Raphael Felizardo | Universidade Federal de São Paulo (UNIFESP) (original) (raw)

Papers by Raphael Felizardo

Pharmacological Research, 2019

The bacteria community living in the gut maintains a symbiotic relationship with the host and its... more The bacteria community living in the gut maintains a symbiotic relationship with the host and its unbalance has been associated with progression of a wide range of intestinal and extra intestinal conditions. Hypertension and chronic kidney disease (CKD) are closely associated diseases with high incidence rates all over the world. Increasing data have supported the involvement of gut microbiome in the blood pressure regulation and the impairment of CKD prognosis. In hypertension, the reduced number of short-chain fatty acids (SCFAs) producing bacteria is associated with modifications in gut environment, involving reduction of the hypoxic gut profile and worsening of the microbial balance, leading to a loss of epithelial barrier integrity, development of gut inflammation and the reduction of SCFAs plasma levels. Those modifications compromise the blood pressure regulation and, as a consequence, favor the end organ damage, also affecting the kidneys. In CKD, impaired renal function leads to accumulation of high levels of uremic toxins that reach the intestine and cause alterations in bacteria composition and fecal metabolite profile, inducing a positive feedback that allows translocation of endotoxins into the bloodstream, which enhances local kidney inflammation and exacerbate kidney injury, compromising even more CKD prognosis. In line with these data, the use of prebiotics, probiotics and fecal microbiota transplantation are becoming efficient therapies to improve the gut dysbiosis aiming hypertension and CKD treatment. This review describes how changes in gut microbiota composition can affect the development of hypertension and the progression of kidney diseases, highlighting the importance of the gut microbial composition uncovering to improve human health maintenance and, especially, for the development of new alternative therapies.

Universidade Federal de São Paulo (UNIFESP), Jul 31, 2017

Introduction: Chronic Kidney Disease (CKD) is characterized by structural abnormalities and progr... more Introduction: Chronic Kidney Disease (CKD) is characterized by structural abnormalities and progressive decrease of kidney function until complete loss of filtration capacity. Damages to podocytes, specialized cells involved on filtration process, are one of the major causes that lead to a CKD. The literature comprises a large number of experimental attempts that focus on mitigating the damage to the podocytes. In recent years, many studies point to the gut microbiota as a modulator of intestinal and extraintestinal diseases through the generation of short chain fatty acids (SCFA). It is already known that chronic kidney patients have an imbalanced gut microbiota and lower production of SCFA. Thus, we have explored the role of butyrate, an AGCC able to regulate epigenetic processes and activate G protein coupled receptors (GPR), during the progression of experimental nephropathy induced by adriamycin, focusing mainly on the protection of podocytes. Methods: Wild type mice in a Balb/c background, Gpr109a-/- mice and Gpr109a-/-.Gpr43-/- mice were induced to develop glomerulopathy by a single dose of adriamycin and treated with butyrate. Results: Wild type mice treated with butyrate showed improvement of renal function, associated to a preserved podocyte layer in the glomerular basement membrane and reduction of pro-inflammatory and pro-fibrotic markers in the kidneys. Particularly, butyrate modulated the activity of enzymes involved on epigenetic modifications in the kidneys and changed the frequency of histone markers (H3K9Ac, H3K4me3 and H3K9me3) in the promoter region of the genes encoding synaptopodin, podocin and NEPH-1 in the podocytes. Concomitantly, treatment with butyrate was not sufficient to improve the renal function of Gpr109a -/- mice and Gpr109a-/-.Gpr43-/-mice. Activation of the receptors was associated with the regulations of small GTPases activity Rac1 and Cdc42 and maintenance of the organization of actin filaments in the podocytes grown in vitro. Conclusion: Our results demonstrate that butyrate exerts important effects on podocyte homeostasis during experimental nephropathy through epigenetic and GPR109a receptors-mediated mechanisms.Introdução: A Doença Renal Crônica (DRC) caracteriza-se por anormalidades estruturais e diminuição progressiva da função dos rins até a perda completa da capacidade de filtração. A lesão dos podócitos, células especializadas no processo de filtração glomerular, constitui uma das maiores causas que levam a DRC. Sendo assim, diversos trabalhos tentam procurar alternativas que minimizem os progressivos danos a estas células. Nos últimos anos, muitos trabalhos apontam a microbiota intestinal como moduladora das doenças intestinais e extra-intestinais por meio da geração de ácidos graxos de cadeia curta (AGCC). Hoje sabemos que pacientes renais crônicos possuem desequilíbrio da microbiota intestinal e menor produção de AGCC. Assim, exploramos o papel do butirato, um AGCC com capacidade de regular processos epigenéticos e se ligar a receptores acoplados a proteína G (GPR), em proteger os podócitos durante a progressão da nefropatia experimental induzida pela adriamicina. Metodologia: Camundongos selvagens Balb/c, Gpr109a-/- e Gpr109a-/- x Gpr43-/- foram induzidos a desenvolver glomerulopatia por meio da adriamicina e tratados com butirato. Resultados: Animais selvagens tratados com butirato apresentaram melhora da função da barreira glomerular, associada à preservação da camada de podócitos na membrana basal glomerular e diminuição de marcadores pró-inflamatórios e pró-fibróticos nos rins. Especificamente, o butirato modulou a expressão de enzimas envolvidas em modificações epigenéticas nos rins e alterou a frequência de modificações em histonas (H3K9Ac, H3K4me3 e H3K9me3) na região promotora dos genes que codificam sinaptopodina, podocina e NEPH-1 em podócitos. Paralelamente, o tratamento com butirato não mostrou recuperação da função da barreira glomerular em animais deficientes dos receptores de butirato GPR109a e GPR43. A ativação desses receptores foi associada à modulação da atividade de GTPases como Rac1 e Cdc42 e à manutenção da organização dos filamentos de actina dos pedicelos nos podócitos in vitro. Conclusão: Nossos resultados demonstram que o butirato tem efeitos importantes na homeostase dos podócitos durante a nefropatia experimental por meio de alterações epigenéticas e ativação dos receptores GPR109a.Dados abertos - Sucupira - Teses e dissertações (2017

Copyright © 2014 Matheus Correa-Costa et al. This is an open access article distributed under the... more Copyright © 2014 Matheus Correa-Costa et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Macrophages play a special role in the onset of several diseases, including acute and chronic kidney injuries. In this sense, tubule interstitial nephritis (TIN) represents an underestimated insult, which can be triggered by different stimuli and, in the absence of a proper regulation, can lead to fibrosis deposition. Based on this perception, we evaluated the participation of macrophage recruitment in the development of TIN. Initially, we provided adenine-enriched food toWT and searched formacrophage presence and action in the kidney. Also, a group of animals were depleted of macrophages with the clodronate liposome while receiving adenine-enriched diet. We collected blood and renal tissue from these animals and renal function...

Clinical science (London, England : 1979), Jan 2, 2018

Acute kidney injury (AKI) is considered an inflammatory disease in which Toll-like receptors (TLR... more Acute kidney injury (AKI) is considered an inflammatory disease in which Toll-like receptors (TLRs) signaling pathways play an important role. The activation of TLRs results in production of several inflammatory cytokines leading to further renal damage. In contrast, TLRs are key players on autophagy induction, which is associated with a protective function on cisplatin-induced AKI. Hence, this study aimed evaluate the specific participation of TLR2 and TLR4 molecules on development of cisplatin-induced AKI. Complementarily, we also investigated the link between TLRs and HO-1, a promisor cytoprotective molecule. Firstly, we observed that only the absence of TLR2 but not TLR4 in mice exacerbated the renal dysfunction, tissue injury and mortality rate, even under an immunologically privileged microenvironment. Second, we demonstrated that TLR2 KO mice presented lower expression of autophagy-associated markers when compared with TLR4 KO animals. Similar parameter was confirmed, using t...

Inflammopharmacology, Jan 23, 2017

Inflammatory bowel diseases (IBDs) affect millions of people worldwide and their frequencies in d... more Inflammatory bowel diseases (IBDs) affect millions of people worldwide and their frequencies in developed countries have increased since the twentieth century. In this context, there is an intensive search for therapies that modulate inflammation and provide tissue regeneration in IBDs. Recently, the immunomodulatory activity of adipose tissue-derived mesenchymal stromal cells (ADMSCs) has been demonstrated to play an important role on several immune cells in different conditions of inflammatory and autoimmune diseases. In this study, we explored the immunomodulatory potential of ADMSC in a classical model of DSS-induced colitis. First, we found that treatment of mice with ADMSC ameliorated the severity of DSS-induced colitis, reducing colitis pathological score and preventing colon shortening. Moreover, a prominent reduction of pro-inflammatory cytokines levels (i.e., IFN-γ, TNF-α, IL-6 and MCP-1) was observed in the colon of animals treated with ADMSC. We also observed a significa...

Frontiers in physiology, 2017

The antineoplastic drug cisplatin promotes renal injury, which limits its use. Protocols that red... more The antineoplastic drug cisplatin promotes renal injury, which limits its use. Protocols that reduce renal cisplatin toxicity will allow higher doses to be used in cisplatin treatment. Here, we compare physical exercise and caloric restriction (CR) as protocols to reduce cisplatin renal injury in mice. Male C57BL/6 were divided into four groups: Control, cisplatin, exercise + cisplatin, and 30% CR + cisplatin. Animals were injected with a single dose of cisplatin (20 mg/kg i.p.) and sacrificed 96 h after injection. Quantitative real time PCR, histological analyses, immunohistochemistry, and biochemical measurements were performed to investigate renal injury, necrosis, apoptosis, and inflammatory mechanisms. Both protocols protected against cisplatin renal injury, but CR was more effective in reducing uraemia and renal necrosis. The CR + Cisplatin group exhibited reduced serum IL-1β and TNF-α levels. No differences were noted in the renal mRNA expression of cytokines. Both interventi...

Molecular and cellular biochemistry, Jan 4, 2017

Cisplatin is a drug widely used in chemotherapy that frequently causes severe renal dysfunction. ... more Cisplatin is a drug widely used in chemotherapy that frequently causes severe renal dysfunction. Organic transporters have an important role to control the absorption and excretion of cisplatin in renal cells. Deletion and blockage of kinin B1 receptor has already been show to protect against cisplatin-induced acute kidney injury. To test whether it exerts its protective function by modulating the organic transporters in kidney, we studied kinin B1 receptor knockout mice and treatment with a receptor antagonist at basal state and in presence of cisplatin. Cisplatin administration caused downregulation of renal organic transporters; in B1 receptor knockout mice, this downregulation of organic transporters in kidney was absent; and treatment by a B1 receptor antagonist attenuated the downregulation of the transporter MATE-1. Moreover, kinin B1 receptor deletion and blockage at basal state resulted in higher renal expression of MATE-1. Moreover we observed that kinin B1 receptor deleti...

Frontiers in immunology, 2016

Mesenchymal stromal cells (MSCs) orchestrate tissue repair by releasing cell-derived microvesicle... more Mesenchymal stromal cells (MSCs) orchestrate tissue repair by releasing cell-derived microvesicles (MVs), which, presumably by small RNA species, modulate global gene expression. The knowledge of miRNA/mRNA signatures linked to a reparative status may elucidate some of the molecular events associated with MSC protection. Here, we used a model of cisplatin-induced kidney injury (acute kidney injury) to assess how MSCs or MVs could restore tissue function. MSCs and MVs presented similar protective effects, which were evidenced in vivo and in vitro by modulating apoptosis, inflammation, oxidative stress, and a set of prosurvival molecules. In addition, we observed that miRNAs (i.e., miR-880, miR-141, miR-377, and miR-21) were modulated, thereby showing active participation on regenerative process. Subsequently, we identified that MSC regulates a particular miRNA subset which mRNA targets are associated with Wnt/TGF-β, fibrosis, and epithelial-mesenchymal transition signaling pathways. ...

American Journal of Physiology - Lung Cellular and Molecular Physiology, 2016

Sakuranetin is the main isolate flavonoid from Baccharis retusa ( Asteraceae) leaves and exhibits... more Sakuranetin is the main isolate flavonoid from Baccharis retusa ( Asteraceae) leaves and exhibits anti-inflammatory and antioxidative activities. Acute respiratory distress syndrome is an acute failure of the respiratory system for which effective treatment is urgently necessary. This study investigated the preventive and therapeutic effects of sakuranetin on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Animals were treated with intranasal sakuranetin 30 min before or 6 h after instillation of LPS. Twenty-four hours after ALI was induced, lung function, inflammation, macrophages population markers, collagen fiber deposition, the extent of oxidative stress, and the expression of matrix metalloprotease-9 (MMP-9), tissue inhibitor of MMP-9 (TIMP-1) and NF-κB were evaluated. The animals began to show lung alterations 6 h after LPS instillation, and these changes persisted until 24 h after LPS administration. Preventive and therapeutic treatment with sakuranetin redu...

Clinical & Translational Immunology, 2016

Journal of Stem Cell Research & Therapy, 2014

A considerable number of epileptic patients had become resistant to antiepileptic drugs, justifyi... more A considerable number of epileptic patients had become resistant to antiepileptic drugs, justifying the need for development of new therapeutic strategies to treat epilepsy. The use of mesenchymal stem cells is an innovative and accessible strategy for the treatment of neuronal disorders, due to their involvement in immunoregulatory mechanisms, trophic and anti-apoptotic action. Objective: Based on this evidence, we evaluated the protective effect of mesenchymal cells from adipose tissue (MCAT) by behavioral and inflammatory responses against convulsive seizure induced by maximum electroconvulsive shock (MES). Methods: MCAT cells were transplanted into the hippocampus of adult male mice, and ten days after the transplantation MES stimulation was applied to induce a generalized tonic-clonic seizure. To evaluate the anticonvulsant activity of MCAT cells, we evaluated the parameters involved with: protection and reduction in the duration of tonic phase, reduction in the mortality rate, and alteration in the hippocampal gene expression of IL-1beta, IL-6, IL-4, IL-10, caspase-1, iNOS and TNFα. Results: MCAT cells transplanted into the hippocampus altered the convulsive threshold, showed anticonvulsant effect by protecting from tonic seizures and mortality and reduced the hippocampal expression of transcripts related to inflammatory response such as IL-1beta, IL-6, caspase-1 and iNOS and increased the level of anti-inflammatory interleukin IL-4. Conclusion: The anticonvulsant effects of the MCAT cells on acute convulsive seizure may be related to inhibitory factors and immunomodulatory mechanisms assigned to mesenchymal cells in the hippocampus. These anticonvulsants mechanisms of MCAT cells bring strong therapeutic implications for the control of epileptic seizures.

World Journal of Nephrology, 2014

Disease Models & Mechanisms, 2014

Focal and segmental glomerulosclerosis (FSGS) is one of the most important renal diseases related... more Focal and segmental glomerulosclerosis (FSGS) is one of the most important renal diseases related to end stage renal failure. Bradykinin has been implicated in the pathogenesis of renal inflammation whereas the role of its receptor 2 (B2RBK) in FSGS has not been studied. FSGS was induced in wild type and B2RBK KO mice by a single intravenous injection of Adriamycin (ADM). In order to further modulate the kinin receptors, animals were also treated with B2RBK antagonist HOE-140, and DALBK, B1RBK antagonist. Here, we show that the blockage of B2RBK with HOE-140 protects mice from FSGS development, including podocyte foot process effacement and reestablishment of slit diaphragm-related proteins. However, B2RBK KO mice were not protected from FSGS. These opposite results were due to B1RBK expression. B1RBK was up regulated after ADM injection and it was exacerbated in B2RBK KO animals. Further, HOE-140 treatment down regulated B1RBK receptor. The blockade of B1RBK in B2RBK KO animals pro...

Journal of the American Society of Nephrology : JASN, Jan 14, 2015

Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbio... more Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbiota and have anti-inflammatory and histone deacetylase-inhibiting properties. Recently, a dual relationship between the intestine and kidneys has been unraveled. Therefore, we evaluated the role of SCFA in an AKI model in which the inflammatory process has a detrimental role. We observed that therapy with the three main SCFAs (acetate, propionate, and butyrate) improved renal dysfunction caused by injury. This protection was associated with low levels of local and systemic inflammation, oxidative cellular stress, cell infiltration/activation, and apoptosis. However, it was also associated with an increase in autophagy. Moreover, SCFAs inhibited histone deacetylase activity and modulated the expression levels of enzymes involved in chromatin modification. In vitro analyses showed that SCFAs modulated the inflammatory process, decreasing the maturation of dendritic cells and inhibiting the ...

Frontiers in Immunology

Increasing evidence shows the essential participation of gut microbiota in human health and disea... more Increasing evidence shows the essential participation of gut microbiota in human health and diseases by shaping local and systemic immunity. Despite an accumulating body of studies showing that chronic kidney disease (CKD) is closely associated with disturbances in the composition of gut microbiota, it remains unclear the importance of gut microbiota in the onset and development of CKD. For the purpose of untangling the role of gut microbiota in CKD, gut microbiota was depleted with a pool of broad-spectrum antibiotics in mice submitted to unilateral ureteral obstruction (UUO). Depletion of gut microbiota significantly decreased levels of proinflammatory cytokines and fibrosis markers, attenuating renal injury. Additionally, to study whether the pathogenic role of gut microbiota is dependent of microbial-host crosstalk, we generated mice lacking Myd88 (myeloid differentiation primary response gene 8) expression in intestinal epithelial cells (IECs) and performed UUO. The absence of ...

Inflammatory Bowel Diseases

Background The gut microbiota is a key element to support host homeostasis and the development of... more Background The gut microbiota is a key element to support host homeostasis and the development of the immune system. The relationship between the microbiota and immunity is a 2-way road, in which the microbiota contributes to the development/function of immune cells and immunity can affect the composition of microbes. In this context, natural killer T cells (NKT cells) are distinct T lymphocytes that play a role in gut immunity and are influenced by gut microbes. In our work, we investigated the involvement of invariant NKT cells (iNKT) in intestinal homeostasis. Results We found that iNKT-deficient mice (iNKT-KO) had reduced levels of fecal IgA and an altered composition of the gut microbiota, with increased Bacteroidetes. The absence of iNKT cells also affected TGF-β1 levels and plasma cells, which were significantly reduced in knockout (KO) mice. In addition, when submitted to dextran sodium sulfate colitis, iNKT-KO mice had worsening of colitis when compared with wild-type (WT) ...

International immunopharmacology, Jan 31, 2018

Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public... more Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public health, and their pathophysiology involves immune cells activation, being macrophages one of the main players of both processes. It is suggested that metabolic pathways could contribute to macrophage modulation and phosphatidylinositol‑3 kinase (PI3K) pathway was shown to be activated in kidneys subjected to ischemia and reperfusion as well as unilateral ureteral obstruction (UUO). Although PI3K inhibition is mostly associated with anti-inflammatory response, its use in kidney injuries has been shown controversial results, which indicates the need for further studies. Our aim was to unveil the role of PI3Kγ in macrophage polarization and in kidney diseases development. We analyzed bone-marrow macrophages polarization from wild-type (WT) and PI3Kγ knockout (PI3K KO) animals. We observed increased expression of M1 (CD86, CCR7, iNOS, TNF, CXCL9, CXCL10, IL-12 and IL-23) and decreased of M2...

International Immunopharmacology

Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public... more Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public health, and their pathophysiology involves immune cells activation, being macrophages one of the main players of both processes. It is suggested that metabolic pathways could contribute to macrophage modulation and phosphatidylinositol‑3 kinase (PI3K) pathway was shown to be activated in kidneys subjected to ischemia and reperfusion as well as unilateral ureteral obstruction (UUO). Although PI3K inhibition is mostly associated with anti-inflammatory response, its use in kidney injuries has been shown controversial results, which indicates the need for further studies. Our aim was to unveil the role of PI3Kγ in macrophage polarization and in kidney diseases development. We analyzed bone-marrow macrophages polarization from wild-type (WT) and PI3Kγ knockout (PI3K KO) animals. We observed increased expression of M1 (CD86, CCR7, iNOS, TNF, CXCL9, CXCL10, IL-12 and IL-23) and decreased of M2 (CD206, Arg-1, FIZZ1 and YM1) markers in the lack of PI3Kγ. And this modulation was accompanied by higher levels of inflammatory cytokines in PI3K KO M1 cells. PI3K KO mice had increased M1 in steady state kidneys, and no protection was observed in these mice after acute and chronic kidney insults. On the contrary, they presented higher levels of protein-to-creatinine ratio and Kim-1 expression and increased tubular injury. In conclusion, our findings demonstrated that the lack of PI3Kγ favors M1 macrophages polarization providing an inflammatory-prone environment, which does not prevent kidney diseases progression.

Journal of the American Society of Nephrology : JASN, Jan 14, 2015

Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbio... more Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbiota and have anti-inflammatory and histone deacetylase-inhibiting properties. Recently, a dual relationship between the intestine and kidneys has been unraveled. Therefore, we evaluated the role of SCFA in an AKI model in which the inflammatory process has a detrimental role. We observed that therapy with the three main SCFAs (acetate, propionate, and butyrate) improved renal dysfunction caused by injury. This protection was associated with low levels of local and systemic inflammation, oxidative cellular stress, cell infiltration/activation, and apoptosis. However, it was also associated with an increase in autophagy. Moreover, SCFAs inhibited histone deacetylase activity and modulated the expression levels of enzymes involved in chromatin modification. In vitro analyses showed that SCFAs modulated the inflammatory process, decreasing the maturation of dendritic cells and inhibiting the ...

Pharmacological Research, 2019

The bacteria community living in the gut maintains a symbiotic relationship with the host and its... more The bacteria community living in the gut maintains a symbiotic relationship with the host and its unbalance has been associated with progression of a wide range of intestinal and extra intestinal conditions. Hypertension and chronic kidney disease (CKD) are closely associated diseases with high incidence rates all over the world. Increasing data have supported the involvement of gut microbiome in the blood pressure regulation and the impairment of CKD prognosis. In hypertension, the reduced number of short-chain fatty acids (SCFAs) producing bacteria is associated with modifications in gut environment, involving reduction of the hypoxic gut profile and worsening of the microbial balance, leading to a loss of epithelial barrier integrity, development of gut inflammation and the reduction of SCFAs plasma levels. Those modifications compromise the blood pressure regulation and, as a consequence, favor the end organ damage, also affecting the kidneys. In CKD, impaired renal function leads to accumulation of high levels of uremic toxins that reach the intestine and cause alterations in bacteria composition and fecal metabolite profile, inducing a positive feedback that allows translocation of endotoxins into the bloodstream, which enhances local kidney inflammation and exacerbate kidney injury, compromising even more CKD prognosis. In line with these data, the use of prebiotics, probiotics and fecal microbiota transplantation are becoming efficient therapies to improve the gut dysbiosis aiming hypertension and CKD treatment. This review describes how changes in gut microbiota composition can affect the development of hypertension and the progression of kidney diseases, highlighting the importance of the gut microbial composition uncovering to improve human health maintenance and, especially, for the development of new alternative therapies.

Universidade Federal de São Paulo (UNIFESP), Jul 31, 2017

Introduction: Chronic Kidney Disease (CKD) is characterized by structural abnormalities and progr... more Introduction: Chronic Kidney Disease (CKD) is characterized by structural abnormalities and progressive decrease of kidney function until complete loss of filtration capacity. Damages to podocytes, specialized cells involved on filtration process, are one of the major causes that lead to a CKD. The literature comprises a large number of experimental attempts that focus on mitigating the damage to the podocytes. In recent years, many studies point to the gut microbiota as a modulator of intestinal and extraintestinal diseases through the generation of short chain fatty acids (SCFA). It is already known that chronic kidney patients have an imbalanced gut microbiota and lower production of SCFA. Thus, we have explored the role of butyrate, an AGCC able to regulate epigenetic processes and activate G protein coupled receptors (GPR), during the progression of experimental nephropathy induced by adriamycin, focusing mainly on the protection of podocytes. Methods: Wild type mice in a Balb/c background, Gpr109a-/- mice and Gpr109a-/-.Gpr43-/- mice were induced to develop glomerulopathy by a single dose of adriamycin and treated with butyrate. Results: Wild type mice treated with butyrate showed improvement of renal function, associated to a preserved podocyte layer in the glomerular basement membrane and reduction of pro-inflammatory and pro-fibrotic markers in the kidneys. Particularly, butyrate modulated the activity of enzymes involved on epigenetic modifications in the kidneys and changed the frequency of histone markers (H3K9Ac, H3K4me3 and H3K9me3) in the promoter region of the genes encoding synaptopodin, podocin and NEPH-1 in the podocytes. Concomitantly, treatment with butyrate was not sufficient to improve the renal function of Gpr109a -/- mice and Gpr109a-/-.Gpr43-/-mice. Activation of the receptors was associated with the regulations of small GTPases activity Rac1 and Cdc42 and maintenance of the organization of actin filaments in the podocytes grown in vitro. Conclusion: Our results demonstrate that butyrate exerts important effects on podocyte homeostasis during experimental nephropathy through epigenetic and GPR109a receptors-mediated mechanisms.Introdução: A Doença Renal Crônica (DRC) caracteriza-se por anormalidades estruturais e diminuição progressiva da função dos rins até a perda completa da capacidade de filtração. A lesão dos podócitos, células especializadas no processo de filtração glomerular, constitui uma das maiores causas que levam a DRC. Sendo assim, diversos trabalhos tentam procurar alternativas que minimizem os progressivos danos a estas células. Nos últimos anos, muitos trabalhos apontam a microbiota intestinal como moduladora das doenças intestinais e extra-intestinais por meio da geração de ácidos graxos de cadeia curta (AGCC). Hoje sabemos que pacientes renais crônicos possuem desequilíbrio da microbiota intestinal e menor produção de AGCC. Assim, exploramos o papel do butirato, um AGCC com capacidade de regular processos epigenéticos e se ligar a receptores acoplados a proteína G (GPR), em proteger os podócitos durante a progressão da nefropatia experimental induzida pela adriamicina. Metodologia: Camundongos selvagens Balb/c, Gpr109a-/- e Gpr109a-/- x Gpr43-/- foram induzidos a desenvolver glomerulopatia por meio da adriamicina e tratados com butirato. Resultados: Animais selvagens tratados com butirato apresentaram melhora da função da barreira glomerular, associada à preservação da camada de podócitos na membrana basal glomerular e diminuição de marcadores pró-inflamatórios e pró-fibróticos nos rins. Especificamente, o butirato modulou a expressão de enzimas envolvidas em modificações epigenéticas nos rins e alterou a frequência de modificações em histonas (H3K9Ac, H3K4me3 e H3K9me3) na região promotora dos genes que codificam sinaptopodina, podocina e NEPH-1 em podócitos. Paralelamente, o tratamento com butirato não mostrou recuperação da função da barreira glomerular em animais deficientes dos receptores de butirato GPR109a e GPR43. A ativação desses receptores foi associada à modulação da atividade de GTPases como Rac1 e Cdc42 e à manutenção da organização dos filamentos de actina dos pedicelos nos podócitos in vitro. Conclusão: Nossos resultados demonstram que o butirato tem efeitos importantes na homeostase dos podócitos durante a nefropatia experimental por meio de alterações epigenéticas e ativação dos receptores GPR109a.Dados abertos - Sucupira - Teses e dissertações (2017

Copyright © 2014 Matheus Correa-Costa et al. This is an open access article distributed under the... more Copyright © 2014 Matheus Correa-Costa et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Macrophages play a special role in the onset of several diseases, including acute and chronic kidney injuries. In this sense, tubule interstitial nephritis (TIN) represents an underestimated insult, which can be triggered by different stimuli and, in the absence of a proper regulation, can lead to fibrosis deposition. Based on this perception, we evaluated the participation of macrophage recruitment in the development of TIN. Initially, we provided adenine-enriched food toWT and searched formacrophage presence and action in the kidney. Also, a group of animals were depleted of macrophages with the clodronate liposome while receiving adenine-enriched diet. We collected blood and renal tissue from these animals and renal function...

Clinical science (London, England : 1979), Jan 2, 2018

Acute kidney injury (AKI) is considered an inflammatory disease in which Toll-like receptors (TLR... more Acute kidney injury (AKI) is considered an inflammatory disease in which Toll-like receptors (TLRs) signaling pathways play an important role. The activation of TLRs results in production of several inflammatory cytokines leading to further renal damage. In contrast, TLRs are key players on autophagy induction, which is associated with a protective function on cisplatin-induced AKI. Hence, this study aimed evaluate the specific participation of TLR2 and TLR4 molecules on development of cisplatin-induced AKI. Complementarily, we also investigated the link between TLRs and HO-1, a promisor cytoprotective molecule. Firstly, we observed that only the absence of TLR2 but not TLR4 in mice exacerbated the renal dysfunction, tissue injury and mortality rate, even under an immunologically privileged microenvironment. Second, we demonstrated that TLR2 KO mice presented lower expression of autophagy-associated markers when compared with TLR4 KO animals. Similar parameter was confirmed, using t...

Inflammopharmacology, Jan 23, 2017

Inflammatory bowel diseases (IBDs) affect millions of people worldwide and their frequencies in d... more Inflammatory bowel diseases (IBDs) affect millions of people worldwide and their frequencies in developed countries have increased since the twentieth century. In this context, there is an intensive search for therapies that modulate inflammation and provide tissue regeneration in IBDs. Recently, the immunomodulatory activity of adipose tissue-derived mesenchymal stromal cells (ADMSCs) has been demonstrated to play an important role on several immune cells in different conditions of inflammatory and autoimmune diseases. In this study, we explored the immunomodulatory potential of ADMSC in a classical model of DSS-induced colitis. First, we found that treatment of mice with ADMSC ameliorated the severity of DSS-induced colitis, reducing colitis pathological score and preventing colon shortening. Moreover, a prominent reduction of pro-inflammatory cytokines levels (i.e., IFN-γ, TNF-α, IL-6 and MCP-1) was observed in the colon of animals treated with ADMSC. We also observed a significa...

Frontiers in physiology, 2017

The antineoplastic drug cisplatin promotes renal injury, which limits its use. Protocols that red... more The antineoplastic drug cisplatin promotes renal injury, which limits its use. Protocols that reduce renal cisplatin toxicity will allow higher doses to be used in cisplatin treatment. Here, we compare physical exercise and caloric restriction (CR) as protocols to reduce cisplatin renal injury in mice. Male C57BL/6 were divided into four groups: Control, cisplatin, exercise + cisplatin, and 30% CR + cisplatin. Animals were injected with a single dose of cisplatin (20 mg/kg i.p.) and sacrificed 96 h after injection. Quantitative real time PCR, histological analyses, immunohistochemistry, and biochemical measurements were performed to investigate renal injury, necrosis, apoptosis, and inflammatory mechanisms. Both protocols protected against cisplatin renal injury, but CR was more effective in reducing uraemia and renal necrosis. The CR + Cisplatin group exhibited reduced serum IL-1β and TNF-α levels. No differences were noted in the renal mRNA expression of cytokines. Both interventi...

Molecular and cellular biochemistry, Jan 4, 2017

Cisplatin is a drug widely used in chemotherapy that frequently causes severe renal dysfunction. ... more Cisplatin is a drug widely used in chemotherapy that frequently causes severe renal dysfunction. Organic transporters have an important role to control the absorption and excretion of cisplatin in renal cells. Deletion and blockage of kinin B1 receptor has already been show to protect against cisplatin-induced acute kidney injury. To test whether it exerts its protective function by modulating the organic transporters in kidney, we studied kinin B1 receptor knockout mice and treatment with a receptor antagonist at basal state and in presence of cisplatin. Cisplatin administration caused downregulation of renal organic transporters; in B1 receptor knockout mice, this downregulation of organic transporters in kidney was absent; and treatment by a B1 receptor antagonist attenuated the downregulation of the transporter MATE-1. Moreover, kinin B1 receptor deletion and blockage at basal state resulted in higher renal expression of MATE-1. Moreover we observed that kinin B1 receptor deleti...

Frontiers in immunology, 2016

Mesenchymal stromal cells (MSCs) orchestrate tissue repair by releasing cell-derived microvesicle... more Mesenchymal stromal cells (MSCs) orchestrate tissue repair by releasing cell-derived microvesicles (MVs), which, presumably by small RNA species, modulate global gene expression. The knowledge of miRNA/mRNA signatures linked to a reparative status may elucidate some of the molecular events associated with MSC protection. Here, we used a model of cisplatin-induced kidney injury (acute kidney injury) to assess how MSCs or MVs could restore tissue function. MSCs and MVs presented similar protective effects, which were evidenced in vivo and in vitro by modulating apoptosis, inflammation, oxidative stress, and a set of prosurvival molecules. In addition, we observed that miRNAs (i.e., miR-880, miR-141, miR-377, and miR-21) were modulated, thereby showing active participation on regenerative process. Subsequently, we identified that MSC regulates a particular miRNA subset which mRNA targets are associated with Wnt/TGF-β, fibrosis, and epithelial-mesenchymal transition signaling pathways. ...

American Journal of Physiology - Lung Cellular and Molecular Physiology, 2016

Sakuranetin is the main isolate flavonoid from Baccharis retusa ( Asteraceae) leaves and exhibits... more Sakuranetin is the main isolate flavonoid from Baccharis retusa ( Asteraceae) leaves and exhibits anti-inflammatory and antioxidative activities. Acute respiratory distress syndrome is an acute failure of the respiratory system for which effective treatment is urgently necessary. This study investigated the preventive and therapeutic effects of sakuranetin on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Animals were treated with intranasal sakuranetin 30 min before or 6 h after instillation of LPS. Twenty-four hours after ALI was induced, lung function, inflammation, macrophages population markers, collagen fiber deposition, the extent of oxidative stress, and the expression of matrix metalloprotease-9 (MMP-9), tissue inhibitor of MMP-9 (TIMP-1) and NF-κB were evaluated. The animals began to show lung alterations 6 h after LPS instillation, and these changes persisted until 24 h after LPS administration. Preventive and therapeutic treatment with sakuranetin redu...

Clinical & Translational Immunology, 2016

Journal of Stem Cell Research & Therapy, 2014

A considerable number of epileptic patients had become resistant to antiepileptic drugs, justifyi... more A considerable number of epileptic patients had become resistant to antiepileptic drugs, justifying the need for development of new therapeutic strategies to treat epilepsy. The use of mesenchymal stem cells is an innovative and accessible strategy for the treatment of neuronal disorders, due to their involvement in immunoregulatory mechanisms, trophic and anti-apoptotic action. Objective: Based on this evidence, we evaluated the protective effect of mesenchymal cells from adipose tissue (MCAT) by behavioral and inflammatory responses against convulsive seizure induced by maximum electroconvulsive shock (MES). Methods: MCAT cells were transplanted into the hippocampus of adult male mice, and ten days after the transplantation MES stimulation was applied to induce a generalized tonic-clonic seizure. To evaluate the anticonvulsant activity of MCAT cells, we evaluated the parameters involved with: protection and reduction in the duration of tonic phase, reduction in the mortality rate, and alteration in the hippocampal gene expression of IL-1beta, IL-6, IL-4, IL-10, caspase-1, iNOS and TNFα. Results: MCAT cells transplanted into the hippocampus altered the convulsive threshold, showed anticonvulsant effect by protecting from tonic seizures and mortality and reduced the hippocampal expression of transcripts related to inflammatory response such as IL-1beta, IL-6, caspase-1 and iNOS and increased the level of anti-inflammatory interleukin IL-4. Conclusion: The anticonvulsant effects of the MCAT cells on acute convulsive seizure may be related to inhibitory factors and immunomodulatory mechanisms assigned to mesenchymal cells in the hippocampus. These anticonvulsants mechanisms of MCAT cells bring strong therapeutic implications for the control of epileptic seizures.

World Journal of Nephrology, 2014

Disease Models & Mechanisms, 2014

Focal and segmental glomerulosclerosis (FSGS) is one of the most important renal diseases related... more Focal and segmental glomerulosclerosis (FSGS) is one of the most important renal diseases related to end stage renal failure. Bradykinin has been implicated in the pathogenesis of renal inflammation whereas the role of its receptor 2 (B2RBK) in FSGS has not been studied. FSGS was induced in wild type and B2RBK KO mice by a single intravenous injection of Adriamycin (ADM). In order to further modulate the kinin receptors, animals were also treated with B2RBK antagonist HOE-140, and DALBK, B1RBK antagonist. Here, we show that the blockage of B2RBK with HOE-140 protects mice from FSGS development, including podocyte foot process effacement and reestablishment of slit diaphragm-related proteins. However, B2RBK KO mice were not protected from FSGS. These opposite results were due to B1RBK expression. B1RBK was up regulated after ADM injection and it was exacerbated in B2RBK KO animals. Further, HOE-140 treatment down regulated B1RBK receptor. The blockade of B1RBK in B2RBK KO animals pro...

Journal of the American Society of Nephrology : JASN, Jan 14, 2015

Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbio... more Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbiota and have anti-inflammatory and histone deacetylase-inhibiting properties. Recently, a dual relationship between the intestine and kidneys has been unraveled. Therefore, we evaluated the role of SCFA in an AKI model in which the inflammatory process has a detrimental role. We observed that therapy with the three main SCFAs (acetate, propionate, and butyrate) improved renal dysfunction caused by injury. This protection was associated with low levels of local and systemic inflammation, oxidative cellular stress, cell infiltration/activation, and apoptosis. However, it was also associated with an increase in autophagy. Moreover, SCFAs inhibited histone deacetylase activity and modulated the expression levels of enzymes involved in chromatin modification. In vitro analyses showed that SCFAs modulated the inflammatory process, decreasing the maturation of dendritic cells and inhibiting the ...

Frontiers in Immunology

Increasing evidence shows the essential participation of gut microbiota in human health and disea... more Increasing evidence shows the essential participation of gut microbiota in human health and diseases by shaping local and systemic immunity. Despite an accumulating body of studies showing that chronic kidney disease (CKD) is closely associated with disturbances in the composition of gut microbiota, it remains unclear the importance of gut microbiota in the onset and development of CKD. For the purpose of untangling the role of gut microbiota in CKD, gut microbiota was depleted with a pool of broad-spectrum antibiotics in mice submitted to unilateral ureteral obstruction (UUO). Depletion of gut microbiota significantly decreased levels of proinflammatory cytokines and fibrosis markers, attenuating renal injury. Additionally, to study whether the pathogenic role of gut microbiota is dependent of microbial-host crosstalk, we generated mice lacking Myd88 (myeloid differentiation primary response gene 8) expression in intestinal epithelial cells (IECs) and performed UUO. The absence of ...

Inflammatory Bowel Diseases

Background The gut microbiota is a key element to support host homeostasis and the development of... more Background The gut microbiota is a key element to support host homeostasis and the development of the immune system. The relationship between the microbiota and immunity is a 2-way road, in which the microbiota contributes to the development/function of immune cells and immunity can affect the composition of microbes. In this context, natural killer T cells (NKT cells) are distinct T lymphocytes that play a role in gut immunity and are influenced by gut microbes. In our work, we investigated the involvement of invariant NKT cells (iNKT) in intestinal homeostasis. Results We found that iNKT-deficient mice (iNKT-KO) had reduced levels of fecal IgA and an altered composition of the gut microbiota, with increased Bacteroidetes. The absence of iNKT cells also affected TGF-β1 levels and plasma cells, which were significantly reduced in knockout (KO) mice. In addition, when submitted to dextran sodium sulfate colitis, iNKT-KO mice had worsening of colitis when compared with wild-type (WT) ...

International immunopharmacology, Jan 31, 2018

Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public... more Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public health, and their pathophysiology involves immune cells activation, being macrophages one of the main players of both processes. It is suggested that metabolic pathways could contribute to macrophage modulation and phosphatidylinositol‑3 kinase (PI3K) pathway was shown to be activated in kidneys subjected to ischemia and reperfusion as well as unilateral ureteral obstruction (UUO). Although PI3K inhibition is mostly associated with anti-inflammatory response, its use in kidney injuries has been shown controversial results, which indicates the need for further studies. Our aim was to unveil the role of PI3Kγ in macrophage polarization and in kidney diseases development. We analyzed bone-marrow macrophages polarization from wild-type (WT) and PI3Kγ knockout (PI3K KO) animals. We observed increased expression of M1 (CD86, CCR7, iNOS, TNF, CXCL9, CXCL10, IL-12 and IL-23) and decreased of M2...

International Immunopharmacology

Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public... more Acute kidney injury (AKI) and chronic kidney disease (CKD) are major concerns in worldwide public health, and their pathophysiology involves immune cells activation, being macrophages one of the main players of both processes. It is suggested that metabolic pathways could contribute to macrophage modulation and phosphatidylinositol‑3 kinase (PI3K) pathway was shown to be activated in kidneys subjected to ischemia and reperfusion as well as unilateral ureteral obstruction (UUO). Although PI3K inhibition is mostly associated with anti-inflammatory response, its use in kidney injuries has been shown controversial results, which indicates the need for further studies. Our aim was to unveil the role of PI3Kγ in macrophage polarization and in kidney diseases development. We analyzed bone-marrow macrophages polarization from wild-type (WT) and PI3Kγ knockout (PI3K KO) animals. We observed increased expression of M1 (CD86, CCR7, iNOS, TNF, CXCL9, CXCL10, IL-12 and IL-23) and decreased of M2 (CD206, Arg-1, FIZZ1 and YM1) markers in the lack of PI3Kγ. And this modulation was accompanied by higher levels of inflammatory cytokines in PI3K KO M1 cells. PI3K KO mice had increased M1 in steady state kidneys, and no protection was observed in these mice after acute and chronic kidney insults. On the contrary, they presented higher levels of protein-to-creatinine ratio and Kim-1 expression and increased tubular injury. In conclusion, our findings demonstrated that the lack of PI3Kγ favors M1 macrophages polarization providing an inflammatory-prone environment, which does not prevent kidney diseases progression.

Journal of the American Society of Nephrology : JASN, Jan 14, 2015

Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbio... more Short-chain fatty acids (SCFAs) are fermentation end products produced by the intestinal microbiota and have anti-inflammatory and histone deacetylase-inhibiting properties. Recently, a dual relationship between the intestine and kidneys has been unraveled. Therefore, we evaluated the role of SCFA in an AKI model in which the inflammatory process has a detrimental role. We observed that therapy with the three main SCFAs (acetate, propionate, and butyrate) improved renal dysfunction caused by injury. This protection was associated with low levels of local and systemic inflammation, oxidative cellular stress, cell infiltration/activation, and apoptosis. However, it was also associated with an increase in autophagy. Moreover, SCFAs inhibited histone deacetylase activity and modulated the expression levels of enzymes involved in chromatin modification. In vitro analyses showed that SCFAs modulated the inflammatory process, decreasing the maturation of dendritic cells and inhibiting the ...