Giuseppe Esposito - Academia.edu (original) (raw)

Papers by Giuseppe Esposito

Research paper thumbnail of Oral Immunization with Escherichia coli Nissle 1917 Expressing SARS-CoV-2 Spike Protein Induces Mucosal and Systemic Antibody Responses in Mice

Biomolecules

As of October 2022, the COVID-19 pandemic continues to pose a major public health conundrum, with... more As of October 2022, the COVID-19 pandemic continues to pose a major public health conundrum, with increased rates of symptomatic infections in vaccinated individuals. An ideal vaccine candidate for the prevention of outbreaks should be rapidly scalable, easy to administer, and able to elicit a potent mucosal immunity. Towards this aim, we proposed an engineered Escherichia coli (E. coli) Nissle 1917 (EcN) strain with SARS-CoV-2 spike protein (SP)-coding plasmid, which was able to expose SP on its cellular surface by a hybridization with the adhesin involved in diffuse adherence 1 (AIDA1). In this study, we presented the effectiveness of a 16-week intragastrically administered, engineered EcN in producing specific systemic and mucosal immunoglobulins against SARS-CoV-2 SP in mice. We observed a time-dependent increase in anti-SARS-CoV-2 SP IgG antibodies in the sera at week 4, with a titre that more than doubled by week 12 and a stable circulating titre by week 16 (+309% and +325% vs...

Research paper thumbnail of Oral Adelmidrol Administration Up-Regulates Palmitoylethanolamide Production in Mice Colon and Duodenum through a PPAR-γ Independent Action

Metabolites

Adelmidrol is a promising palmitoylethanolamide (PEA) analog which displayed up-and-coming anti-i... more Adelmidrol is a promising palmitoylethanolamide (PEA) analog which displayed up-and-coming anti-inflammatory properties in several inflammatory conditions. Recent studies demonstrated that Adelmidrol is an in vitro enhancer of PEA endogenous production, through the so called “entourage” effect. The present study investigated the ability of Adelmidrol (1 and 10 mg/Kg per os) to increase the endogenous level of PEA in the duodenum and colon of mice after 21-day oral administration in the presence and absence of PPAR-γ inhibitor (1 mg/kg). The level of PEA was analyzed by HPLC-MS. The expression of PEA-related enzymatic machinery was evaluated by western blot and RT-PCR analysis. Our findings demonstrated that Adelmidrol significantly increased PEA levels in the duodenum and colon in a dose/time-dependent manner. We also revealed that Adelmidrol up regulated the enzymatic machinery responsible for PEA metabolism and catabolism. Interestingly, the use of the selective irreversible PPAR-...

Research paper thumbnail of N-Palmitoyl-D-Glucosamine Inhibits TLR-4/NLRP3 and Improves DNBS-Induced Colon Inflammation through a PPAR-α-Dependent Mechanism

Biomolecules

Similar to canine inflammatory enteropathy, inflammatory bowel disease (IBD) is a chronic idiopat... more Similar to canine inflammatory enteropathy, inflammatory bowel disease (IBD) is a chronic idiopathic condition characterized by remission periods and recurrent flares in which diarrhea, visceral pain, rectal bleeding/bloody stools, and weight loss are the main clinical symptoms. Intestinal barrier function alterations often persist in the remission phase of the disease without ongoing inflammatory processes. However, current therapies include mainly anti-inflammatory compounds that fail to promote functional symptoms-free disease remission, urging new drug discoveries to handle patients during this step of the disease. ALIAmides (ALIA, autacoid local injury antagonism) are bioactive fatty acid amides that recently gained attention because of their involvement in the control of inflammatory response, prompting the use of these molecules as plausible therapeutic strategies in the treatment of several chronic inflammatory conditions. N-palmitoyl-D-glucosamine (PGA), an under-researched...

Research paper thumbnail of Sleeve Gastrectomy-Induced Body Mass Index Reduction Increases the Intensity of Taste Perception’s and Reduces Bitter-Induced Pleasantness in Severe Obesity

Journal of Clinical Medicine

Background: The sense of taste is involved in food behavior and may drive food choices, likely co... more Background: The sense of taste is involved in food behavior and may drive food choices, likely contributing to obesity. Differences in taste preferences have been reported in normal-weight as compared to obese subjects. Changes in taste perception with an increased sweet-induced sensitivity have been reported in surgically treated obese patients, but data regarding the perception of basic tastes yielded conflicting results. We aimed to evaluate basic taste identification, induced perception, and pleasantness in normal-weight controls and obese subjects before and after bariatric surgery. Methods: Severe obese and matched normal weight subjects underwent a standardized spit test to evaluate sweet, bitter, salty, umami, and sour taste identification, induced perception, and pleasantness. A subset of obese subjects were also studied before and 12 months after sleeve gastrectomy. Results: No significant differences in basic taste-induced perceptions were observed, although a higher numb...

Research paper thumbnail of A Palmitoylethanolamide Producing Lactobacillus paracasei Improves Clostridium difficile Toxin A-Induced Colitis

Frontiers in Pharmacology

Genetically engineered probiotics, able to in situ deliver therapeutically active compounds while... more Genetically engineered probiotics, able to in situ deliver therapeutically active compounds while restoring gut eubiosis, could represent an attractive therapeutic alternative in Clostridium difficile infection (CDI). Palmitoylethanolamide is an endogenous lipid able to exert immunomodulatory activities and restore epithelial barrier integrity in human models of colitis, by binding the peroxisome proliferator–activated receptor-α (PPARα). The aim of this study was to explore the efficacy of a newly designed PEA-producing probiotic (pNAPE-LP) in a mice model of C. difficile toxin A (TcdA)-induced colitis. The human N-acyl-phosphatidylethanolamine-specific phospholipase D (NAPE-PLD), a key enzyme involved in the synthesis of PEA, was cloned and expressed in a Lactobacillus paracasei that was intragastrically administered to mice 7 days prior the induction of the colitis. Bacteria carrying the empty vector served as negative controls (pLP).In the presence of palmitate, pNAPE-LP was abl...

Research paper thumbnail of Next-Generation Probiotics for Inflammatory Bowel Disease

International Journal of Molecular Sciences

Engineered probiotics represent a cutting-edge therapy in intestinal inflammatory disease (IBD). ... more Engineered probiotics represent a cutting-edge therapy in intestinal inflammatory disease (IBD). Genetically modified bacteria have provided a new strategy to release therapeutically operative molecules in the intestine and have grown into promising new therapies for IBD. Current IBD treatments, such as corticosteroids and immunosuppressants, are associated with relevant side effects and a significant proportion of patients are dependent on these therapies, thus exposing them to the risk of relevant long-term side effects. Discovering new and effective therapeutic strategies is a worldwide goal in this research field and engineered probiotics could potentially provide a viable solution. This review aims at describing the proceeding of bacterial engineering and how genetically modified probiotics may represent a promising new biotechnological approach in IBD treatment.

Research paper thumbnail of Nutraceuticals and Diet Supplements in Crohn’s Disease: A General Overview of the Most Promising Approaches in the Clinic

Foods

Crohn’s disease (CD) is a chronic inflammatory gastrointestinal disorder requiring lifelong medic... more Crohn’s disease (CD) is a chronic inflammatory gastrointestinal disorder requiring lifelong medications. The currently approved drugs for CD are associated with relevant side effects and several studies suggest an increased use of nutraceuticals among CD patients, seeking for what is perceived as a more “natural” approach in controlling this highly morbid condition. Nutraceuticals are foods or foods’ components with beneficial health properties that could aid in CD treatment for their anti-inflammatory, analgesic and immunoregulatory activities that come along with safety, high tolerability, easy availability and affordability. Depending on their biological effect, nutraceuticals’ support could be employed in different subsets of CD patients, both those with active disease, as adjunctive immunomodulatory therapies, and/or in quiescent disease to provide symptomatic relief in patients with residual functional symptoms. Despite the increasing interest of the general public, both limit...

Research paper thumbnail of Phytotherapics inCOVID19: Why palmitoylethanolamide?

Phytotherapy Research, 2020

At present, googling the search terms “COVID‐19” and “Functional foods” yields nearly 500,000,000... more At present, googling the search terms “COVID‐19” and “Functional foods” yields nearly 500,000,000 hits, witnessing the growing interest of the scientific community and the general public in the role of nutrition and nutraceuticals during the COVID‐19 pandemic. Many compounds have been proposed as phytotherapics in the prevention and/or treatment of COVID‐19. The extensive interest of the general public and the enormous social media coverage on this topic urges the scientific community to address the question of whether which nutraceuticals can actually be employed in preventing and treating this newly described coronavirus‐related disease. Recently, the Canadian biotech pharma company “FSD Pharma” received the green light from the Food and Drug Administration to design a proof‐of‐concept study evaluating the effects of ultramicronized palmitoylethanolamide (PEA) in COVID‐19 patients. The story of PEA as a nutraceutical to prevent and treat infectious diseases dates back to the 1970s where the molecule was branded under the name Impulsin and was used for its immunomodulatory properties in influenza virus infection. The present paper aims at analyzing the potential of PEA as a nutraceutical and the previous evidence suggesting its anti‐inflammatory and immunomodulatory properties in infectious and respiratory diseases and how these could translate to COVID‐19 care.

Research paper thumbnail of Lathyrus sativus diamine oxidase reduces Clostridium difficile toxin A‐induced toxicity in Caco‐2 cells by rescuing RhoA‐GTPase and inhibiting pp38‐MAPK / NF‐κB / HIF ‐1α activation

Phytotherapy Research, 2020

Clostridium difficile toxin A (TcdA) impairs the intestinal epithelial barrier, increasing the mu... more Clostridium difficile toxin A (TcdA) impairs the intestinal epithelial barrier, increasing the mucosa permeability and triggering a robust inflammatory response. Lathyrus sativus diamino oxidase (LSAO) is a nutraceutical compound successfully used in various gastrointestinal dysfunctions. Here, we evaluated the LSAO (0.004–0.4 μM) ability to counter TcdA‐induced (30 ng/mL) toxicity and damage in Caco‐2 cells, investigating its possible mechanism of action. LSAO has improved the transepithelial electrical resistance (TEER) score and increased cell viability in TcdA‐treated cells, significantly rescuing the protein expression of Ras homolog family members, A‐GTPase (RhoA‐GTPase), occludin, and zonula occludens‐1 (ZO‐1). LSAO has also exhibited an anti‐apoptotic effect by inhibiting the TcdA‐induced expression of Bcl‐2‐associated X protein (Bax), p50 nuclear factor‐kappa‐B (p50), p65nuclear factor‐kappa‐B (p65), and hypoxia‐inducible transcription factor‐1 alpha (HIF‐1α), and the release of tumor necrosis factor‐alpha (TNF‐α), interleukin‐6 (IL‐6), and vascular endothelial growth factor (VEGF) in the cell milieu. Our data showed that LSAO exerts a protective effect on TcdA‐induced toxicity in Caco‐2 cells, placing itself as an interesting nutraceutical to supplement the current treatment of the Clostridium difficile infections.

Research paper thumbnail of Lathyrus sativus diamine oxidase counteracts histamine‐induced cell proliferation, migration and pro‐angiogenic mediators release in human colon adenocarcinoma cell line Caco‐2

Phytotherapy Research, 2019

Because histamine is a modulator of cancer cell proliferation and invasiveness, this study aimed ... more Because histamine is a modulator of cancer cell proliferation and invasiveness, this study aimed at investigating the effect of Lathyrus sativus-derived diamine oxidase (LSAO) and its mechanism of action on Caco-2 cell line, considering that LSAO catalizes the oxidative deamination of histamine to the corresponding aldehyde, NH 3 and H 2 O 2. Histamine (0.01-1 μM) caused a proliferative effect on Caco-2 cells promoting cell migration, invasion and nitric oxide and vascular endothelial growth factor release. Histamine (1 μM) stimulus also down regulated occludin expression, favouring up regulation of pro-proliferative nuclear protein Ki67. Incubation with LSAO (0.004-0.4 μM) resulted in a significant inhibition of histamine-induced effects. LSAO rescued occludin expression and down regulated Ki67, and it inhibited histamine-induced increase of both MMP-2 and 9 expression. Histamine effects were mediated by RhoA-GTP down regulation and inversely related to phospho-p38MAPK/p50/65 up regulation. These effects were counteracted by LSAO incubation. Histamine catabolism by LSAO accounts for a significant down regulation of proliferation and invasiveness of Caco-2 cells. This study highlights the importance to control histamine levels in contrasting pro-angiogenic and metastatization capability of colon cancer cells and expands the knowledge about the diamine oxidase from L. sativus seeding as a phytotherapeutic approach for colon cancer.

Research paper thumbnail of Play in advance against neurodegeneration: exploring enteric glial cells in gut-brain axis during neurodegenerative diseases

Expert Review of Clinical Pharmacology, 2019

Introduction: New investigations have shown that "activated" enteric glial cells (EGCs), astrocyt... more Introduction: New investigations have shown that "activated" enteric glial cells (EGCs), astrocyte-like cells of the enteric nervous system (ENS), represent a possible extra-CNS trigger point of the neurodegenerative processes in impaired intestinal permeability conditions. The early modulation of enteric glia-mediated neuroinflammation might optimize neuroprotective treatments outcomes currently used in neurodegenerative diseases. Areas covered: We discussed recent clinical and preclinical data existing on Pubmed database, concerning the glial role in neurodegeneration. We focused on the gut as possible "entrance door" for endoluminal neurotoxic agents that induce neurological impairments during leaky gut conditions. Moreover, we reviewed the paradigmatic studies linking the leaky gut-A c c e p t e d M a n u s c r i p t Information Classification: General induced priming of EGCs to the induction of late neurodegenerative processes in Parkinson's disease and other neurodegenerative disorders. Expert opinion: The previous appearance of neuropathological markers in the ENS emphasizes the extra-CNS origin of neurodegenerative disorders, by directing their therapies toward peripheral management of neurodegeneration. In light of the EGCs changes resulting from a switchon of activated phenotype in leaky gut syndrome, EGCs sampling could be predictive for neuropathological conditions detection, anticipating their symptomatic manifestation in the CNS.

Research paper thumbnail of Endocannabinoids in the treatment of gasytrointestinal inflammation and symptoms

Current opinion in pharmacology, Jan 12, 2018

The evolving policies regarding the use of therapeutic Cannabis have steadily increased the publi... more The evolving policies regarding the use of therapeutic Cannabis have steadily increased the public interest in its use as a complementary and alternative medicine in several disorders, including inflammatory bowel disease. Endocannabinoids represent both an appealing therapeutic strategy and a captivating scientific dilemma. Results from clinical trials have to be carefully interpreted owing to possible reporting-biases related to cannabinoids psychotropic effects. Moreover, discriminating between symptomatic improvement and the real gain on the underlying inflammatory process is often challenging. This review summarizes the advances and latest discovery in this ever-changing field of investigation, highlighting the main limitations in the current use of these drugs in clinical practice and the possible future perspectives to overcome these flaws.

Research paper thumbnail of Palmitoylethanolamide Modulates Inflammation-Associated Vascular Endothelial Growth Factor (VEGF) Signaling via the Akt/mTOR Pathway in a Selective Peroxisome Proliferator-Activated Receptor Alpha (PPAR-α)-Dependent Manner

PLOS ONE, 2016

Background and Aim Angiogenesis is emerging as a pivotal process in chronic inflammatory patholog... more Background and Aim Angiogenesis is emerging as a pivotal process in chronic inflammatory pathologies, promoting immune infiltration and prompting carcinogenesis. Ulcerative Colitis (UC) and Crohn's Disease (CD) represent paradigmatic examples of intestinal chronic inflammatory conditions in which the process of neovascularization correlates with the severity and progression of the diseases. Molecules able to target the angiogenesis have thus the potential to synergistically affect the disease course. Beyond its anti-inflammatory effect, palmitoylethanolamide (PEA) is able to reduce angiogenesis in several chronic inflammatory conditions, but no data about its anti-angiogenic activity in colitis have been produced, yet. Methods The effects of PEA on inflammation-associated angiogenesis in mice with dextran sulphate sodium (DSS)-induced colitis and in patients with UC were assessed. The release of Vascular Endothelial Growth Factor (VEGF), the hemoglobin tissue content, the expression of CD31 and of phosphatidylinositol 3-kinase/Akt/mammalian-target-of-rapamycin (mTOR) signaling axis were all evaluated in the presence of different concentrations of PEA and concomitant administration of PPAR-α and-γ antagonists.

Research paper thumbnail of Cannabidiol in medicine: a review of its therapeutic potential in CNS disorders

Phytotherapy Research, 2009

Cannabidiol (CBD) is the main non-psychotropic component of the glandular hairs of Cannabis sativ... more Cannabidiol (CBD) is the main non-psychotropic component of the glandular hairs of Cannabis sativa. It displays a plethora of actions including anticonvulsive, sedative, hypnotic, antipsychotic, antiinflammatory and neuroprotective properties. However, it is well established that CBD produces its biological effects without exerting significant intrinsic activity upon cannabinoid receptors. For this reason, CBD lacks the unwanted psychotropic effects characteristic of marijuana derivatives, so representing one of the bioactive constituents of Cannabis sativa with the highest potential for therapeutic use. The present review reports the pharmacological profile of CBD and summarizes results from preclinical and clinical studies utilizing CBD, alone or in combination with other phytocannabinoids, for the treatment of a number of CNS disorders.

Research paper thumbnail of Cannabidiol in Inflammatory Bowel Diseases: A Brief Overview

Phytotherapy Research, 2012

This minireview highlights the importance of cannabidiol (CBD) as a promising drug for the therap... more This minireview highlights the importance of cannabidiol (CBD) as a promising drug for the therapy of inflammatory bowel diseases (IBD). Actual pharmacological treatments for IBD should be enlarged toward the search for lowtoxicityand low-cost drugs that may be given alone or in combination with the conventional anti-IBD drugs to increase their efficacy in the therapy of relapsing forms of colitis. In the past, Cannabis preparations have been considered new promising pharmacological tools in view of their anti-inflammatory role in IBD as well as other gut disturbances. However, their use in the clinical therapy has been strongly limited by their psychotropic effects. CBD is a very promising compound since it shares the typical cannabinoid beneficial effects on gut lacking any psychotropic effects. For years, its activity has been enigmatic for gastroenterologists and pharmacologists, but now it is evident that this compound may interact at extra-cannabinoid system receptor sites, such as peroxisome proliferator-activated receptor-gamma. This strategic interaction makes CBD as a potential candidate for the development of a new class of anti-IBD drugs.

Research paper thumbnail of The astroglial-derived S100β protein stimulates the expression of nitric oxide synthase in rodent macrophages through p38 MAP kinase activation

Life Sciences, 2006

S100h is an astroglial-derived Ca 2+-binding protein having neurotrophic role on neurons and glia... more S100h is an astroglial-derived Ca 2+-binding protein having neurotrophic role on neurons and glial cells. An aberrant S100h production has been observed in neurodegenerative disease, as Alzheimer's disease and Down syndrome. S100h is responsible to start up a gliotic reaction by the release of pro-inflammatory mediators, including nitric oxide (NO) and cytokines from microglia and astrocytes, which are, in turn, deleterious for neurons. Interestingly, pro-inflammatory effect of S100h seems not be restricted into the brain. Macrophages play a pivotal role in inflammatory diseases, occurring both in the brain and in the periphery. In this study, we tested the hypothesis that S100h may affect macrophage functions, amplifying thus the inflammatory process. Our results demonstrate that S100h stimulates both NO production and iNOS protein transcription and expression in J774 and rat peritoneal macrophages. NO production was concentration and time-dependently inhibited by two iNOS inhibitors, l-NAME and SMT. We also demonstrated that S100h induced oxidative stress by increasing H 2 O 2 production and lipid peroxidation of cell membrane in both macrophage types. The pro-oxidant potential of S100h activates p38 MAP kinase (MAPK), which has been described to directly activate NF-nB. In our study, SB203580, a p38 MAPK inhibitor, and two NF-nB inhibitors, TLCK and BAY 11-7082, decreased both NO production and iNOS protein transcription and expression in S100h-stimulated J774 and peritoneal rat macrophages. Moreover, additional studies demonstrated that S100h affected also TNF-a protein expression in J774 macrophages. In conclusion, our results highlight the potential role of S100h during an inflammatory scenario identifying macrophages as a novel S100h-responsive cell-type.

Research paper thumbnail of Local administration of WIN 55,212-2 reduces chronic granuloma-associated angiogenesis in rat by inhibiting NF-κB activation

Journal of Molecular Medicine, 2007

Chronic inflammation is often associated with granuloma formation that is a hallmark of many huma... more Chronic inflammation is often associated with granuloma formation that is a hallmark of many human diseases. The transcription factor nuclear factor-kappa B (NF-κB) plays a central role in this process by regulating the expression of several pro-inflammatory genes. Cannabinoids (CBs) from Cannabis sativa L. exert a large number of biological effects including anti-inflammatory and anti-angiogenic effects. In this study, we investigated the role of CBs on granuloma formation induced by λcarrageenin-soaked sponge implant in rat. Our results show that local administration of WIN 55,212-2, a CB 1 /CB 2 agonist, given daily or at time of implantation significantly decreased weight and neo-angiogenesis in granuloma tissue and inhibited nuclear factor-kappa B (NF-κB)/DNA binding that was associated with a reduced inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), tumor

Research paper thumbnail of S100B induces tau protein hyperphosphorylation via Dickopff-1 up-regulation and disrupts the Wnt pathway in human neural stem cells

Journal of Cellular and Molecular Medicine, 2008

Previous studies suggest that levels of the astrocyte-derived S100B protein, such as those occurr... more Previous studies suggest that levels of the astrocyte-derived S100B protein, such as those occurring in brain extracellular spaces consequent to persistent astroglial activation, may have a pathogenetic role in Alzheimer's disease (AD). Although S100B was reported to promote ␤ amyloid precursor protein overexpression, no clear mechanistic relationship between S100B and formation of neurofibrillary tangles (NFTs) is established. This in vitro study has been aimed at investigating whether S100B is able to disrupt Wnt pathway and lead to tau protein hyperphosphorylation. Utilizing Western blot, electrophoretic mobility shift assay, supershift and reverse transcriptase-polymerase chain reaction techniques, it has been demonstrated that micromolar S100B concentrations stimulate c-Jun N-terminal kinase (JNK) phosphorylation through the receptor for advanced glycation ending products, and subsequently activate nuclear AP-1/cJun transcription, in cultured human neural stem cells. In addition, as revealed by Western blot, small interfering RNA and immunofluorescence analysis, S100B-induced JNK activation increased expression of Dickopff-1 that, in turn, promoted glycogen synthase kinase 3␤ phosphorylation and ␤-catenin degradation, causing canonical Wnt pathway disruption and tau protein hyperphosphorylation. These findings propose a previously unrecognized link between S100B and tau hyperphosphorylation, suggesting S100B can contribute to NFT formation in AD and in all other conditions in which neuroinflammation may have a crucial role.

Research paper thumbnail of Selective CB2 up-regulation in women affected by endometrial inflammation

Journal of Cellular and Molecular Medicine, 2008

Endometritis is defined as an inflammation of the endometrial mucosa of the uterus. In endometrit... more Endometritis is defined as an inflammation of the endometrial mucosa of the uterus. In endometritis large amounts of toxic mediators, including nitric oxide (NO) are released by inflammatory cells. As a consequence of nitric oxide-dependent injury, the cells respond by triggering protective mechanisms, by changing the endocannabinoid system (ECS) which comprises both CB1 and CB2 cannabinoid receptors and their endogenous ligands. The aim of our study was to seek out evidence for the presence of cannabinoid receptors in inflammatory endometrial tissue as well as for their potential role in endometrial inflammation. Our results showed a selective up-regulation of both transcription and expression of CB2 receptors in biopsies from women affected by endometrial inflammation compared to healthy women. The experiments with the nitric oxide-donor S-Nitroso-L-Glutathione (GSNO) suggest that such a selective up-regulation may be related to the nitric oxide release occurring during endometrial inflammation. In addition, we demonstrated an increase in chymase expression, a marker of mast cells, in biopsies of women affected by endometritis. Therefore our results support the hypothesis that the up-regulation of CB2 occurs mainly on mast cells and that it might tend to sensitize these cells to the anti-inflammatory effect exerted by endogenous cannabinoids by binding their receptor and thus preventing the mast cell degranulation and the release of pro-inflammatory mediators. In conclusion, we believe that the selective CB2 up-regulation might play a role as a novel prognostic factor in endometrial inflammation.

Research paper thumbnail of The Endocannabinoid System Protects Rat Glioma Cells Against HIV-1 Tat Protein-induced Cytotoxicity

Journal of Biological Chemistry, 2002

Cannabinoids modulate nitric oxide (NO) levels in cells of the central nervous system. Here we st... more Cannabinoids modulate nitric oxide (NO) levels in cells of the central nervous system. Here we studied the effect of cannabinoid CB 1 and CB 2 receptor agonists on the release of NO and cell toxicity induced by the human immuno-deficiency virus-1 Tat protein (HIV-1 Tat) in rat glioma C6 cells. The CB 1 and CB 2 agonist WIN 55,212-2 inhibited the expression of inducible NO synthase (iNOS) and NO release caused by treatment of C6 cells with HIV-1 Tat and interferon-␥ (IFN-␥). The effect of WIN 55,212-2 was uniquely due to CB 1 receptors, as shown by experiments carried out with selective CB 1 and CB 2 receptor agonists and antagonists. CB 1 receptor stimulation also inhibited HIV-1 Tat ؉ IFN-␥-induced and NO-mediated cell toxicity. Moreover, cell treatment with HIV-1 Tat ؉ IFN-␥ induced a significant inhibition of CB 1 , but not CB 2 , receptor expression. This effect was mimicked by the NO donor GSNO, suggesting that the inhibition of CB 1 expression was due to HIV-1 Tat ؉ IFN-␥-induced NO overexpression. HIV-1 Tat ؉ IFN-␥ treatment also induced a significant inhibition of the uptake of the endocannabinoid anandamide by C6 cells with no effect on anandamide hydrolysis. These findings show that the endocannabinoid system, through the modulation of the L-arginine/NO pathway, reduces HIV-1 Tat-induced cytotoxicity, and is itself regulated by HIV-1 Tat.

Research paper thumbnail of Oral Immunization with Escherichia coli Nissle 1917 Expressing SARS-CoV-2 Spike Protein Induces Mucosal and Systemic Antibody Responses in Mice

Biomolecules

As of October 2022, the COVID-19 pandemic continues to pose a major public health conundrum, with... more As of October 2022, the COVID-19 pandemic continues to pose a major public health conundrum, with increased rates of symptomatic infections in vaccinated individuals. An ideal vaccine candidate for the prevention of outbreaks should be rapidly scalable, easy to administer, and able to elicit a potent mucosal immunity. Towards this aim, we proposed an engineered Escherichia coli (E. coli) Nissle 1917 (EcN) strain with SARS-CoV-2 spike protein (SP)-coding plasmid, which was able to expose SP on its cellular surface by a hybridization with the adhesin involved in diffuse adherence 1 (AIDA1). In this study, we presented the effectiveness of a 16-week intragastrically administered, engineered EcN in producing specific systemic and mucosal immunoglobulins against SARS-CoV-2 SP in mice. We observed a time-dependent increase in anti-SARS-CoV-2 SP IgG antibodies in the sera at week 4, with a titre that more than doubled by week 12 and a stable circulating titre by week 16 (+309% and +325% vs...

Research paper thumbnail of Oral Adelmidrol Administration Up-Regulates Palmitoylethanolamide Production in Mice Colon and Duodenum through a PPAR-γ Independent Action

Metabolites

Adelmidrol is a promising palmitoylethanolamide (PEA) analog which displayed up-and-coming anti-i... more Adelmidrol is a promising palmitoylethanolamide (PEA) analog which displayed up-and-coming anti-inflammatory properties in several inflammatory conditions. Recent studies demonstrated that Adelmidrol is an in vitro enhancer of PEA endogenous production, through the so called “entourage” effect. The present study investigated the ability of Adelmidrol (1 and 10 mg/Kg per os) to increase the endogenous level of PEA in the duodenum and colon of mice after 21-day oral administration in the presence and absence of PPAR-γ inhibitor (1 mg/kg). The level of PEA was analyzed by HPLC-MS. The expression of PEA-related enzymatic machinery was evaluated by western blot and RT-PCR analysis. Our findings demonstrated that Adelmidrol significantly increased PEA levels in the duodenum and colon in a dose/time-dependent manner. We also revealed that Adelmidrol up regulated the enzymatic machinery responsible for PEA metabolism and catabolism. Interestingly, the use of the selective irreversible PPAR-...

Research paper thumbnail of N-Palmitoyl-D-Glucosamine Inhibits TLR-4/NLRP3 and Improves DNBS-Induced Colon Inflammation through a PPAR-α-Dependent Mechanism

Biomolecules

Similar to canine inflammatory enteropathy, inflammatory bowel disease (IBD) is a chronic idiopat... more Similar to canine inflammatory enteropathy, inflammatory bowel disease (IBD) is a chronic idiopathic condition characterized by remission periods and recurrent flares in which diarrhea, visceral pain, rectal bleeding/bloody stools, and weight loss are the main clinical symptoms. Intestinal barrier function alterations often persist in the remission phase of the disease without ongoing inflammatory processes. However, current therapies include mainly anti-inflammatory compounds that fail to promote functional symptoms-free disease remission, urging new drug discoveries to handle patients during this step of the disease. ALIAmides (ALIA, autacoid local injury antagonism) are bioactive fatty acid amides that recently gained attention because of their involvement in the control of inflammatory response, prompting the use of these molecules as plausible therapeutic strategies in the treatment of several chronic inflammatory conditions. N-palmitoyl-D-glucosamine (PGA), an under-researched...

Research paper thumbnail of Sleeve Gastrectomy-Induced Body Mass Index Reduction Increases the Intensity of Taste Perception’s and Reduces Bitter-Induced Pleasantness in Severe Obesity

Journal of Clinical Medicine

Background: The sense of taste is involved in food behavior and may drive food choices, likely co... more Background: The sense of taste is involved in food behavior and may drive food choices, likely contributing to obesity. Differences in taste preferences have been reported in normal-weight as compared to obese subjects. Changes in taste perception with an increased sweet-induced sensitivity have been reported in surgically treated obese patients, but data regarding the perception of basic tastes yielded conflicting results. We aimed to evaluate basic taste identification, induced perception, and pleasantness in normal-weight controls and obese subjects before and after bariatric surgery. Methods: Severe obese and matched normal weight subjects underwent a standardized spit test to evaluate sweet, bitter, salty, umami, and sour taste identification, induced perception, and pleasantness. A subset of obese subjects were also studied before and 12 months after sleeve gastrectomy. Results: No significant differences in basic taste-induced perceptions were observed, although a higher numb...

Research paper thumbnail of A Palmitoylethanolamide Producing Lactobacillus paracasei Improves Clostridium difficile Toxin A-Induced Colitis

Frontiers in Pharmacology

Genetically engineered probiotics, able to in situ deliver therapeutically active compounds while... more Genetically engineered probiotics, able to in situ deliver therapeutically active compounds while restoring gut eubiosis, could represent an attractive therapeutic alternative in Clostridium difficile infection (CDI). Palmitoylethanolamide is an endogenous lipid able to exert immunomodulatory activities and restore epithelial barrier integrity in human models of colitis, by binding the peroxisome proliferator–activated receptor-α (PPARα). The aim of this study was to explore the efficacy of a newly designed PEA-producing probiotic (pNAPE-LP) in a mice model of C. difficile toxin A (TcdA)-induced colitis. The human N-acyl-phosphatidylethanolamine-specific phospholipase D (NAPE-PLD), a key enzyme involved in the synthesis of PEA, was cloned and expressed in a Lactobacillus paracasei that was intragastrically administered to mice 7 days prior the induction of the colitis. Bacteria carrying the empty vector served as negative controls (pLP).In the presence of palmitate, pNAPE-LP was abl...

Research paper thumbnail of Next-Generation Probiotics for Inflammatory Bowel Disease

International Journal of Molecular Sciences

Engineered probiotics represent a cutting-edge therapy in intestinal inflammatory disease (IBD). ... more Engineered probiotics represent a cutting-edge therapy in intestinal inflammatory disease (IBD). Genetically modified bacteria have provided a new strategy to release therapeutically operative molecules in the intestine and have grown into promising new therapies for IBD. Current IBD treatments, such as corticosteroids and immunosuppressants, are associated with relevant side effects and a significant proportion of patients are dependent on these therapies, thus exposing them to the risk of relevant long-term side effects. Discovering new and effective therapeutic strategies is a worldwide goal in this research field and engineered probiotics could potentially provide a viable solution. This review aims at describing the proceeding of bacterial engineering and how genetically modified probiotics may represent a promising new biotechnological approach in IBD treatment.

Research paper thumbnail of Nutraceuticals and Diet Supplements in Crohn’s Disease: A General Overview of the Most Promising Approaches in the Clinic

Foods

Crohn’s disease (CD) is a chronic inflammatory gastrointestinal disorder requiring lifelong medic... more Crohn’s disease (CD) is a chronic inflammatory gastrointestinal disorder requiring lifelong medications. The currently approved drugs for CD are associated with relevant side effects and several studies suggest an increased use of nutraceuticals among CD patients, seeking for what is perceived as a more “natural” approach in controlling this highly morbid condition. Nutraceuticals are foods or foods’ components with beneficial health properties that could aid in CD treatment for their anti-inflammatory, analgesic and immunoregulatory activities that come along with safety, high tolerability, easy availability and affordability. Depending on their biological effect, nutraceuticals’ support could be employed in different subsets of CD patients, both those with active disease, as adjunctive immunomodulatory therapies, and/or in quiescent disease to provide symptomatic relief in patients with residual functional symptoms. Despite the increasing interest of the general public, both limit...

Research paper thumbnail of Phytotherapics inCOVID19: Why palmitoylethanolamide?

Phytotherapy Research, 2020

At present, googling the search terms “COVID‐19” and “Functional foods” yields nearly 500,000,000... more At present, googling the search terms “COVID‐19” and “Functional foods” yields nearly 500,000,000 hits, witnessing the growing interest of the scientific community and the general public in the role of nutrition and nutraceuticals during the COVID‐19 pandemic. Many compounds have been proposed as phytotherapics in the prevention and/or treatment of COVID‐19. The extensive interest of the general public and the enormous social media coverage on this topic urges the scientific community to address the question of whether which nutraceuticals can actually be employed in preventing and treating this newly described coronavirus‐related disease. Recently, the Canadian biotech pharma company “FSD Pharma” received the green light from the Food and Drug Administration to design a proof‐of‐concept study evaluating the effects of ultramicronized palmitoylethanolamide (PEA) in COVID‐19 patients. The story of PEA as a nutraceutical to prevent and treat infectious diseases dates back to the 1970s where the molecule was branded under the name Impulsin and was used for its immunomodulatory properties in influenza virus infection. The present paper aims at analyzing the potential of PEA as a nutraceutical and the previous evidence suggesting its anti‐inflammatory and immunomodulatory properties in infectious and respiratory diseases and how these could translate to COVID‐19 care.

Research paper thumbnail of Lathyrus sativus diamine oxidase reduces Clostridium difficile toxin A‐induced toxicity in Caco‐2 cells by rescuing RhoA‐GTPase and inhibiting pp38‐MAPK / NF‐κB / HIF ‐1α activation

Phytotherapy Research, 2020

Clostridium difficile toxin A (TcdA) impairs the intestinal epithelial barrier, increasing the mu... more Clostridium difficile toxin A (TcdA) impairs the intestinal epithelial barrier, increasing the mucosa permeability and triggering a robust inflammatory response. Lathyrus sativus diamino oxidase (LSAO) is a nutraceutical compound successfully used in various gastrointestinal dysfunctions. Here, we evaluated the LSAO (0.004–0.4 μM) ability to counter TcdA‐induced (30 ng/mL) toxicity and damage in Caco‐2 cells, investigating its possible mechanism of action. LSAO has improved the transepithelial electrical resistance (TEER) score and increased cell viability in TcdA‐treated cells, significantly rescuing the protein expression of Ras homolog family members, A‐GTPase (RhoA‐GTPase), occludin, and zonula occludens‐1 (ZO‐1). LSAO has also exhibited an anti‐apoptotic effect by inhibiting the TcdA‐induced expression of Bcl‐2‐associated X protein (Bax), p50 nuclear factor‐kappa‐B (p50), p65nuclear factor‐kappa‐B (p65), and hypoxia‐inducible transcription factor‐1 alpha (HIF‐1α), and the release of tumor necrosis factor‐alpha (TNF‐α), interleukin‐6 (IL‐6), and vascular endothelial growth factor (VEGF) in the cell milieu. Our data showed that LSAO exerts a protective effect on TcdA‐induced toxicity in Caco‐2 cells, placing itself as an interesting nutraceutical to supplement the current treatment of the Clostridium difficile infections.

Research paper thumbnail of Lathyrus sativus diamine oxidase counteracts histamine‐induced cell proliferation, migration and pro‐angiogenic mediators release in human colon adenocarcinoma cell line Caco‐2

Phytotherapy Research, 2019

Because histamine is a modulator of cancer cell proliferation and invasiveness, this study aimed ... more Because histamine is a modulator of cancer cell proliferation and invasiveness, this study aimed at investigating the effect of Lathyrus sativus-derived diamine oxidase (LSAO) and its mechanism of action on Caco-2 cell line, considering that LSAO catalizes the oxidative deamination of histamine to the corresponding aldehyde, NH 3 and H 2 O 2. Histamine (0.01-1 μM) caused a proliferative effect on Caco-2 cells promoting cell migration, invasion and nitric oxide and vascular endothelial growth factor release. Histamine (1 μM) stimulus also down regulated occludin expression, favouring up regulation of pro-proliferative nuclear protein Ki67. Incubation with LSAO (0.004-0.4 μM) resulted in a significant inhibition of histamine-induced effects. LSAO rescued occludin expression and down regulated Ki67, and it inhibited histamine-induced increase of both MMP-2 and 9 expression. Histamine effects were mediated by RhoA-GTP down regulation and inversely related to phospho-p38MAPK/p50/65 up regulation. These effects were counteracted by LSAO incubation. Histamine catabolism by LSAO accounts for a significant down regulation of proliferation and invasiveness of Caco-2 cells. This study highlights the importance to control histamine levels in contrasting pro-angiogenic and metastatization capability of colon cancer cells and expands the knowledge about the diamine oxidase from L. sativus seeding as a phytotherapeutic approach for colon cancer.

Research paper thumbnail of Play in advance against neurodegeneration: exploring enteric glial cells in gut-brain axis during neurodegenerative diseases

Expert Review of Clinical Pharmacology, 2019

Introduction: New investigations have shown that "activated" enteric glial cells (EGCs), astrocyt... more Introduction: New investigations have shown that "activated" enteric glial cells (EGCs), astrocyte-like cells of the enteric nervous system (ENS), represent a possible extra-CNS trigger point of the neurodegenerative processes in impaired intestinal permeability conditions. The early modulation of enteric glia-mediated neuroinflammation might optimize neuroprotective treatments outcomes currently used in neurodegenerative diseases. Areas covered: We discussed recent clinical and preclinical data existing on Pubmed database, concerning the glial role in neurodegeneration. We focused on the gut as possible "entrance door" for endoluminal neurotoxic agents that induce neurological impairments during leaky gut conditions. Moreover, we reviewed the paradigmatic studies linking the leaky gut-A c c e p t e d M a n u s c r i p t Information Classification: General induced priming of EGCs to the induction of late neurodegenerative processes in Parkinson's disease and other neurodegenerative disorders. Expert opinion: The previous appearance of neuropathological markers in the ENS emphasizes the extra-CNS origin of neurodegenerative disorders, by directing their therapies toward peripheral management of neurodegeneration. In light of the EGCs changes resulting from a switchon of activated phenotype in leaky gut syndrome, EGCs sampling could be predictive for neuropathological conditions detection, anticipating their symptomatic manifestation in the CNS.

Research paper thumbnail of Endocannabinoids in the treatment of gasytrointestinal inflammation and symptoms

Current opinion in pharmacology, Jan 12, 2018

The evolving policies regarding the use of therapeutic Cannabis have steadily increased the publi... more The evolving policies regarding the use of therapeutic Cannabis have steadily increased the public interest in its use as a complementary and alternative medicine in several disorders, including inflammatory bowel disease. Endocannabinoids represent both an appealing therapeutic strategy and a captivating scientific dilemma. Results from clinical trials have to be carefully interpreted owing to possible reporting-biases related to cannabinoids psychotropic effects. Moreover, discriminating between symptomatic improvement and the real gain on the underlying inflammatory process is often challenging. This review summarizes the advances and latest discovery in this ever-changing field of investigation, highlighting the main limitations in the current use of these drugs in clinical practice and the possible future perspectives to overcome these flaws.

Research paper thumbnail of Palmitoylethanolamide Modulates Inflammation-Associated Vascular Endothelial Growth Factor (VEGF) Signaling via the Akt/mTOR Pathway in a Selective Peroxisome Proliferator-Activated Receptor Alpha (PPAR-α)-Dependent Manner

PLOS ONE, 2016

Background and Aim Angiogenesis is emerging as a pivotal process in chronic inflammatory patholog... more Background and Aim Angiogenesis is emerging as a pivotal process in chronic inflammatory pathologies, promoting immune infiltration and prompting carcinogenesis. Ulcerative Colitis (UC) and Crohn's Disease (CD) represent paradigmatic examples of intestinal chronic inflammatory conditions in which the process of neovascularization correlates with the severity and progression of the diseases. Molecules able to target the angiogenesis have thus the potential to synergistically affect the disease course. Beyond its anti-inflammatory effect, palmitoylethanolamide (PEA) is able to reduce angiogenesis in several chronic inflammatory conditions, but no data about its anti-angiogenic activity in colitis have been produced, yet. Methods The effects of PEA on inflammation-associated angiogenesis in mice with dextran sulphate sodium (DSS)-induced colitis and in patients with UC were assessed. The release of Vascular Endothelial Growth Factor (VEGF), the hemoglobin tissue content, the expression of CD31 and of phosphatidylinositol 3-kinase/Akt/mammalian-target-of-rapamycin (mTOR) signaling axis were all evaluated in the presence of different concentrations of PEA and concomitant administration of PPAR-α and-γ antagonists.

Research paper thumbnail of Cannabidiol in medicine: a review of its therapeutic potential in CNS disorders

Phytotherapy Research, 2009

Cannabidiol (CBD) is the main non-psychotropic component of the glandular hairs of Cannabis sativ... more Cannabidiol (CBD) is the main non-psychotropic component of the glandular hairs of Cannabis sativa. It displays a plethora of actions including anticonvulsive, sedative, hypnotic, antipsychotic, antiinflammatory and neuroprotective properties. However, it is well established that CBD produces its biological effects without exerting significant intrinsic activity upon cannabinoid receptors. For this reason, CBD lacks the unwanted psychotropic effects characteristic of marijuana derivatives, so representing one of the bioactive constituents of Cannabis sativa with the highest potential for therapeutic use. The present review reports the pharmacological profile of CBD and summarizes results from preclinical and clinical studies utilizing CBD, alone or in combination with other phytocannabinoids, for the treatment of a number of CNS disorders.

Research paper thumbnail of Cannabidiol in Inflammatory Bowel Diseases: A Brief Overview

Phytotherapy Research, 2012

This minireview highlights the importance of cannabidiol (CBD) as a promising drug for the therap... more This minireview highlights the importance of cannabidiol (CBD) as a promising drug for the therapy of inflammatory bowel diseases (IBD). Actual pharmacological treatments for IBD should be enlarged toward the search for lowtoxicityand low-cost drugs that may be given alone or in combination with the conventional anti-IBD drugs to increase their efficacy in the therapy of relapsing forms of colitis. In the past, Cannabis preparations have been considered new promising pharmacological tools in view of their anti-inflammatory role in IBD as well as other gut disturbances. However, their use in the clinical therapy has been strongly limited by their psychotropic effects. CBD is a very promising compound since it shares the typical cannabinoid beneficial effects on gut lacking any psychotropic effects. For years, its activity has been enigmatic for gastroenterologists and pharmacologists, but now it is evident that this compound may interact at extra-cannabinoid system receptor sites, such as peroxisome proliferator-activated receptor-gamma. This strategic interaction makes CBD as a potential candidate for the development of a new class of anti-IBD drugs.

Research paper thumbnail of The astroglial-derived S100β protein stimulates the expression of nitric oxide synthase in rodent macrophages through p38 MAP kinase activation

Life Sciences, 2006

S100h is an astroglial-derived Ca 2+-binding protein having neurotrophic role on neurons and glia... more S100h is an astroglial-derived Ca 2+-binding protein having neurotrophic role on neurons and glial cells. An aberrant S100h production has been observed in neurodegenerative disease, as Alzheimer's disease and Down syndrome. S100h is responsible to start up a gliotic reaction by the release of pro-inflammatory mediators, including nitric oxide (NO) and cytokines from microglia and astrocytes, which are, in turn, deleterious for neurons. Interestingly, pro-inflammatory effect of S100h seems not be restricted into the brain. Macrophages play a pivotal role in inflammatory diseases, occurring both in the brain and in the periphery. In this study, we tested the hypothesis that S100h may affect macrophage functions, amplifying thus the inflammatory process. Our results demonstrate that S100h stimulates both NO production and iNOS protein transcription and expression in J774 and rat peritoneal macrophages. NO production was concentration and time-dependently inhibited by two iNOS inhibitors, l-NAME and SMT. We also demonstrated that S100h induced oxidative stress by increasing H 2 O 2 production and lipid peroxidation of cell membrane in both macrophage types. The pro-oxidant potential of S100h activates p38 MAP kinase (MAPK), which has been described to directly activate NF-nB. In our study, SB203580, a p38 MAPK inhibitor, and two NF-nB inhibitors, TLCK and BAY 11-7082, decreased both NO production and iNOS protein transcription and expression in S100h-stimulated J774 and peritoneal rat macrophages. Moreover, additional studies demonstrated that S100h affected also TNF-a protein expression in J774 macrophages. In conclusion, our results highlight the potential role of S100h during an inflammatory scenario identifying macrophages as a novel S100h-responsive cell-type.

Research paper thumbnail of Local administration of WIN 55,212-2 reduces chronic granuloma-associated angiogenesis in rat by inhibiting NF-κB activation

Journal of Molecular Medicine, 2007

Chronic inflammation is often associated with granuloma formation that is a hallmark of many huma... more Chronic inflammation is often associated with granuloma formation that is a hallmark of many human diseases. The transcription factor nuclear factor-kappa B (NF-κB) plays a central role in this process by regulating the expression of several pro-inflammatory genes. Cannabinoids (CBs) from Cannabis sativa L. exert a large number of biological effects including anti-inflammatory and anti-angiogenic effects. In this study, we investigated the role of CBs on granuloma formation induced by λcarrageenin-soaked sponge implant in rat. Our results show that local administration of WIN 55,212-2, a CB 1 /CB 2 agonist, given daily or at time of implantation significantly decreased weight and neo-angiogenesis in granuloma tissue and inhibited nuclear factor-kappa B (NF-κB)/DNA binding that was associated with a reduced inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), tumor

Research paper thumbnail of S100B induces tau protein hyperphosphorylation via Dickopff-1 up-regulation and disrupts the Wnt pathway in human neural stem cells

Journal of Cellular and Molecular Medicine, 2008

Previous studies suggest that levels of the astrocyte-derived S100B protein, such as those occurr... more Previous studies suggest that levels of the astrocyte-derived S100B protein, such as those occurring in brain extracellular spaces consequent to persistent astroglial activation, may have a pathogenetic role in Alzheimer's disease (AD). Although S100B was reported to promote ␤ amyloid precursor protein overexpression, no clear mechanistic relationship between S100B and formation of neurofibrillary tangles (NFTs) is established. This in vitro study has been aimed at investigating whether S100B is able to disrupt Wnt pathway and lead to tau protein hyperphosphorylation. Utilizing Western blot, electrophoretic mobility shift assay, supershift and reverse transcriptase-polymerase chain reaction techniques, it has been demonstrated that micromolar S100B concentrations stimulate c-Jun N-terminal kinase (JNK) phosphorylation through the receptor for advanced glycation ending products, and subsequently activate nuclear AP-1/cJun transcription, in cultured human neural stem cells. In addition, as revealed by Western blot, small interfering RNA and immunofluorescence analysis, S100B-induced JNK activation increased expression of Dickopff-1 that, in turn, promoted glycogen synthase kinase 3␤ phosphorylation and ␤-catenin degradation, causing canonical Wnt pathway disruption and tau protein hyperphosphorylation. These findings propose a previously unrecognized link between S100B and tau hyperphosphorylation, suggesting S100B can contribute to NFT formation in AD and in all other conditions in which neuroinflammation may have a crucial role.

Research paper thumbnail of Selective CB2 up-regulation in women affected by endometrial inflammation

Journal of Cellular and Molecular Medicine, 2008

Endometritis is defined as an inflammation of the endometrial mucosa of the uterus. In endometrit... more Endometritis is defined as an inflammation of the endometrial mucosa of the uterus. In endometritis large amounts of toxic mediators, including nitric oxide (NO) are released by inflammatory cells. As a consequence of nitric oxide-dependent injury, the cells respond by triggering protective mechanisms, by changing the endocannabinoid system (ECS) which comprises both CB1 and CB2 cannabinoid receptors and their endogenous ligands. The aim of our study was to seek out evidence for the presence of cannabinoid receptors in inflammatory endometrial tissue as well as for their potential role in endometrial inflammation. Our results showed a selective up-regulation of both transcription and expression of CB2 receptors in biopsies from women affected by endometrial inflammation compared to healthy women. The experiments with the nitric oxide-donor S-Nitroso-L-Glutathione (GSNO) suggest that such a selective up-regulation may be related to the nitric oxide release occurring during endometrial inflammation. In addition, we demonstrated an increase in chymase expression, a marker of mast cells, in biopsies of women affected by endometritis. Therefore our results support the hypothesis that the up-regulation of CB2 occurs mainly on mast cells and that it might tend to sensitize these cells to the anti-inflammatory effect exerted by endogenous cannabinoids by binding their receptor and thus preventing the mast cell degranulation and the release of pro-inflammatory mediators. In conclusion, we believe that the selective CB2 up-regulation might play a role as a novel prognostic factor in endometrial inflammation.

Research paper thumbnail of The Endocannabinoid System Protects Rat Glioma Cells Against HIV-1 Tat Protein-induced Cytotoxicity

Journal of Biological Chemistry, 2002

Cannabinoids modulate nitric oxide (NO) levels in cells of the central nervous system. Here we st... more Cannabinoids modulate nitric oxide (NO) levels in cells of the central nervous system. Here we studied the effect of cannabinoid CB 1 and CB 2 receptor agonists on the release of NO and cell toxicity induced by the human immuno-deficiency virus-1 Tat protein (HIV-1 Tat) in rat glioma C6 cells. The CB 1 and CB 2 agonist WIN 55,212-2 inhibited the expression of inducible NO synthase (iNOS) and NO release caused by treatment of C6 cells with HIV-1 Tat and interferon-␥ (IFN-␥). The effect of WIN 55,212-2 was uniquely due to CB 1 receptors, as shown by experiments carried out with selective CB 1 and CB 2 receptor agonists and antagonists. CB 1 receptor stimulation also inhibited HIV-1 Tat ؉ IFN-␥-induced and NO-mediated cell toxicity. Moreover, cell treatment with HIV-1 Tat ؉ IFN-␥ induced a significant inhibition of CB 1 , but not CB 2 , receptor expression. This effect was mimicked by the NO donor GSNO, suggesting that the inhibition of CB 1 expression was due to HIV-1 Tat ؉ IFN-␥-induced NO overexpression. HIV-1 Tat ؉ IFN-␥ treatment also induced a significant inhibition of the uptake of the endocannabinoid anandamide by C6 cells with no effect on anandamide hydrolysis. These findings show that the endocannabinoid system, through the modulation of the L-arginine/NO pathway, reduces HIV-1 Tat-induced cytotoxicity, and is itself regulated by HIV-1 Tat.