Alanood Algarni | Nottingham Trent University (original) (raw)
Papers by Alanood Algarni
International Journal of General Medicine
Background: Pain is a global health issue that affects an individual's quality of life. Its allev... more Background: Pain is a global health issue that affects an individual's quality of life. Its alleviation and management will enhance patients' experience. Community pharmacists can help manage pain severity through their valuable roles in medical teams and by managing the consequences of pain. Objective: This study aimed to evaluate community pharmacists' knowledge and attitudes toward pain and pain management in Saudi Arabia. Methods: A cross-sectional study was performed to evaluate community pharmacists' knowledge and attitudes toward pain and pain management in Saudi Arabia. Pharmacists aged ≥ 21 years, with a degree in pharmacy were included in this study. Each respondent participated in an online survey covering cancer-oriented pain and assessment of pain; pharmacology; abuse of substances; and physical dependence. An independent t-test and One-way ANOVA, with least significant difference as a post-hoc test, were employed, in addition to the General Linear Regression Model using Main Effect as the model. Results: This study revealed that the pain-related knowledge and attitude among community pharmacists in Saudi Arabia were inadequate. Age (p = 0.003), work experience (p = 0.036), nature of work (p = 0.001), and work location (p = 0.003) were determined as significant factors affecting their overall knowledge and attitude toward pain. Conclusion: Overall, attempts to expand community pharmacists' knowledge and foster an appropriate attitude toward pain management among them in Saudi Arabia are highly recommended. Additional academic courses, studies, and tailored neuroscience courses will improve their awareness and knowledge of pain and pain management.
Cancer Treatment and Research Communications
International Journal Of Pharmaceutical Research And Allied Sciences
King Khalid University Journal of Health Sciences
Clinical Pharmacology: Advances and Applications
Obesity and overweight pose a threat to health and are more common than undernutrition among adul... more Obesity and overweight pose a threat to health and are more common than undernutrition among adults. It is categorized by fat accumulation and a body mass index (BMI) of > 30. A significant increase in worldwide obesity has been ongoing over several decades. Over the past few years, several strategies have been followed for weight management and to counteract the increasing prevalence of the disease; however, room for improvement with pharmacological options still exists. This review aimed to digest selected past clinical and experimental studies and understand the role of semaglutide treatment for obesity. Methods: Articles related to the clinical uses of semaglutide, mechanism of action, pharmacokinetics, pharmacodynamics, and side effects of the drug were identified. Only studies with human subjects who used Semaglutide for obesity management were included and assessed. Results: Semaglutide promotes weight loss via appetite and hunger suppression, decreases energy intake, controls eating, and depresses the relative fondness for fatty, energy-dense foods. Moreover, the relationship between obesity and Semaglutide has been widely investigated, and most studies reveal the efficacy of Semaglutide on weight loss. Overall, the pharmacokinetics of semaglutide shows a drop in glycosylated hemoglobin A1c (HbA1c) and total body weight. The usual adverse effects observed in patients treated with Semaglutide include gastrointestinal adverse events, like nausea, vomiting, diarrhea, constipation, and abdominal cramps. Conclusion: The findings from the review suggest that semaglutide appears to be beneficial, most notably in its contribution to weight reduction.
Evidence-Based Complementary and Alternative Medicine, 2022
Many medicinal plants have been utilized for centuries despite the lack of scientific evidence of... more Many medicinal plants have been utilized for centuries despite the lack of scientific evidence of their therapeutic effects. This study evaluated the phytochemical and dual biological profiling, namely, antibacterial and cytotoxic properties, of three plant species, namely, Tribulus terrestris L., Typha domingensis Pers., and Ricinus communis L., in order to explore potential relationships (if any) with their ethnopharmacological uses. GC-MS was used to achieve phytochemical screening of two plant extracts (T. terrestris and T. domingensis). The primary chemicals detected in varying amounts in both extracts were siloxane derivatives, fatty acid esters, diisooctyl phthalate, phytosterol, and aromatic acid esters. According to the findings, the major component detected in both extracts was 1,2-benzenedicarboxylic acid and diisooctyl ester (antibacterial and antifungal). T. domingensis contained a low level of benzoic acid, methyl ester (antibacterial). Both extracts included stigmaste...
Purpose: In a sample of Middle Eastern population, we sought to evaluate whether survival outcome... more Purpose: In a sample of Middle Eastern population, we sought to evaluate whether survival outcomes for mCRC patients treated with systemic chemotherapy (SC) and, with and without biologic therapies (bevacizumab and cetuximab) are different between left and right-sided tumors. Methods: Ministry of National Guard- Health Affairs (MNG-HA) Cancer registry data was utilized to identify patients diagnosed with CRC between 2013 and 2016. Kaplan-Meier method and porosity score adjusted Cox proportional hazard models were used to assess survival for right and left-sided mCRC with and with BT. Results: A total of 196 mCRC patients with mean age of 64 years old, 57.65% male diagnosed from January 1, 2013, through December 31, 2016. The overall median survival for left-sided was significantly higher than right-sided mCRC tumors (P 0.03). mCRC patients treated with SC+BT were associated with decreased mortality only among patients with left-sided mCRC compared to right-sided mCRC (HR, 0.21; 95% ...
World Journal of Pediatrics, 2022
Background Multisystem inflammatory syndrome in children (MIS-C) is a serious health condition th... more Background Multisystem inflammatory syndrome in children (MIS-C) is a serious health condition that develops from and is linked to coronavirus disease 2019. MIS-C is considered a multi-organ dysfunction involving cardiac, renal, respiratory, hematologic, gastrointestinal and neurological symptoms and groups of signs and symptoms such as rash or bilateral nonpurulent conjunctivitis, hypotension or shock and acute gastrointestinal problems, which require immediate therapeutic intervention to prevent the aggravation of the patient's health condition. MIS-C is relatively new in the field of evidence-based medicine; however, there are several clinical guidelines for good clinical practice. For every disorder, the guidelines have different suggestions. Hence, based on the current status of the evidence, recommendations have been combined to form a unified guideline for therapeutic management. Methods This paper compares and evaluates the current MIS-C-specific clinical practice guidelines (namely, American
Several studies have reported that the multifunctional enzyme tissue transglutaminase (TG2) play ... more Several studies have reported that the multifunctional enzyme tissue transglutaminase (TG2) play important roles in neurite outgrowth and modulation of neuronal cell survival. Recently, TG2 enzymatic activity has been shown to be regulated by protein kinases such as PKA and PKC, modulating through G-protein coupled receptors (GPCRs). However, the regulation of TG2 following stimulation of GPCRs coupled to PKC and PKA activation are not fully understood. In neuronal cells, the neurotrophic factors; pituitary adenylate cyclase activating polypeptide (PACAP) and nerve growth factor (NGF) promote the differentiation, maturation, neurite outgrowth and survival of neurons via their GPCR's; the pituitary adenylate cyclase type 1 receptor (PAC1) and the tyrosine kinase receptor (TrKA) including multiple protein kinase signalling pathways. As some of these kinase pathways are associated with modulation of intracellular TG2 activity, it is conceivable to hypothesised that PACAP and NGF di...
Molecules, 2021
Breast cancer is a complex and multi-drug resistant (MDR) disease, which could result in the fail... more Breast cancer is a complex and multi-drug resistant (MDR) disease, which could result in the failure of many chemotherapeutic clinical agents. Discovering effective molecules from natural products or by derivatization from known compounds is the interest of many research studies. The first objective of the present study is to investigate the cytotoxic combinatorial, chemosensitizing, and apoptotic effects of an isatin derived compound (5,5-diphenylimidazolidine-2,4-dione conjugated with 5-substituted isatin, named HAA2021 in the present study) against breast cancer cells (MCF7) and breast cancer cells resistant to doxorubicin (MCF7/ADR) when combined with doxorubicin. The second objective is to investigate the binding mode of HAA2021 withP-glycoprotein (P-gp) and heat shock protein 90 (Hsp90), and to determine whether their co-inhibition by HAA2021 contribute to the increase of the chemosensitization of MCF7/ADR cells to doxorubicin. The combination of HAA2021, at non-toxic doses, w...
Journal of Pharmaceutical Research International
Aim: In this study, we aimed to investigate the incidence rate, risk factors, and mortality rates... more Aim: In this study, we aimed to investigate the incidence rate, risk factors, and mortality rates in patients with early-stage breast cancer using anti-HER2 (Human epidermal growth factor receptor-2) treatment. Patients and Methods: A total of 106 patients diagnosed with human epidermal growth factor 2 (HER2)-positive early-stage breast cancer and receiving anti-HER2 treatment at King Abdulaziz Medical City (KAMC) from 2015 to 2019 were included in the analysis to assess the incidence of cardiotoxicity was collected as a retrospective study. Univariate and multivariate analyses as well as multiple exact logistic regression analysis were conducted to understand the relationships between the left ventricular ejection fraction (LVEF) and treatment combinations and comorbidities Results: The LVEF measurements using an echocardiography method at the baseline (before any treatment) and during the anti-HER2 therapy were assessed. The results suggest that the higher the drug combination, th...
Journal of Molecular Structure
Archives of Toxicology
Organophosphate compounds (OPs) induce both acute and delayed neurotoxic effects, the latter of w... more Organophosphate compounds (OPs) induce both acute and delayed neurotoxic effects, the latter of which is believed to involve their interaction with proteins other than acetylcholinesterase. However, few OP-binding proteins have been identified that may have a direct role in OP-induced delayed neurotoxicity. Given their ability to disrupt Ca2+ homeostasis, a key aim of the current work was to investigate the effects of sub-lethal neurite outgrowth inhibitory levels of OPs on the Ca2+-dependent enzyme tissue transglutaminase (TG2). At 1–10 µM, the OPs phenyl saligenin phosphate (PSP) and chlorpyrifos oxon (CPO) had no effect cell viability but induced concentration-dependent decreases in neurite outgrowth in differentiating N2a neuroblastoma cells. The activity of TG2 increased in cell lysates of differentiating cells exposed for 24 h to PSP and chlorpyrifos oxon CPO (10 µM), as determined by biotin-cadaverine incorporation assays. Exposure to both OPs (3 and/or 10 µM) also enhanced i...
European Journal of Pharmacology
NGF (nerve growth factor) and tissue transglutaminase (TG2) play important roles in neurite outgr... more NGF (nerve growth factor) and tissue transglutaminase (TG2) play important roles in neurite outgrowth and modulation of neuronal cell survival. In this study, we investigated the regulation of TG2 transamidase activity by NGF in retinoic acid-induced differentiating mouse N2a and human SH-SY5Y neuroblastoma cells. TG2 transamidase activity was determined using an amine incorporation and a peptide cross linking assay. In situ TG2 activity was assessed by visualising the incorporation of biotin-X-cadaverine using confocal microscopy. The role of TG2 in NGF-induced cytoprotection and neurite outgrowth was investigated by monitoring hypoxia-induced cell death and appearance of axonal-like processes, respectively. The amine incorporation and protein crosslinking activity of TG2 increased in a time and concentration-dependent manner following stimulation with NGF in N2a and SH-SY5Y cells. NGF mediated increases in TG2 activity were abolished by the TG2 inhibitors Z-DON (Z-ZON-Val-Pro-Leu-OMe; Benzyloxycarbonyl-(6-Diazo-5-oxonorleucinyl)-L-valinyl-L-prolinyl-L-leucinmethylester) and R283 (1,3,dimethyl-2[2-oxo-propyl]thio)imidazole chloride) and by pharmacological inhibition of extracellular signal-regulated kinases 1 and 2 (ERK1/2), protein kinase B (PKB) and protein kinase C (PKC), and removal of extracellular Ca 2+. Fluorescence microscopy demonstrated NGF induced in situ TG2 activity. TG2 inhibition blocked NGFinduced attenuation of hypoxia-induced cell death and neurite outgrowth in both cell lines. Together, these results demonstrate that NGF stimulates TG2 transamidase activity via a ERK1/2, PKB and PKC-dependent pathway in differentiating mouse N2a and human SH-SY5Y neuroblastoma cells. Furthermore, NGF-induced cytoprotection and neurite outgrowth are dependent upon TG2. These results suggest a novel and important role of TG2 in the cellular functions of NGF.
Biochemical Pharmacology, Mar 1, 2017
The PAC1 receptor and tissue transglutaminase (TG2) play important roles in neurite outgrowth and... more The PAC1 receptor and tissue transglutaminase (TG2) play important roles in neurite outgrowth and modulation of neuronal cell survival. In this study, we investigated the regulation of TG2 activity by the PAC1 receptor in retinoic acid-induced differentiating N2a neuroblastoma cells. TG2 transamidase activity was determined using an amine incorporation and a peptide cross linking assay. In situ TG2 activity was assessed by visualising the incorporation of biotin-X-cadaverine using confocal microscopy. TG2 phosphorylation was monitored via immunoprecipitation and Western blotting. The role of TG2 in PAC1 receptor-induced cytoprotection and neurite outgrowth was investigated by monitoring hypoxia-induced cell death and appearance of axonal-like processes, respectively. The amine incorporation and protein crosslinking activity of TG2 increased in a time and concentration-dependent manner following stimulation with pituitary adenylate cyclase-activating polypeptide-27 (PACAP-27). PACAP-27 mediated increases in TG2 activity were abolished by the TG2 inhibitors Z-DON and R283 and by pharmacological inhibition of protein kinase A (KT 5720 and Rp-cAMPs), protein kinase C (Ro 31-8220), MEK1/2 (PD 98059), and removal of extracellular Ca 2+. Fluorescence microscopy demonstrated PACAP-27 induced in situ TG2 activity. TG2 inhibition blocked PACAP-27 induced attenuation of hypoxia-induced cell death and outgrowth of axon-like processes. TG2 activation and cytoprotection were also observed in human SH-SY5Y cells. Together, these results demonstrate that TG2 activity was stimulated downstream of the PAC1 receptor via a multi protein kinase dependent pathway. Furthermore, PAC1 receptor-induced cytoprotection and neurite outgrowth are dependent upon TG2. These results highlight the importance of TG2 in the cellular functions of the PAC1 receptor.
International Journal of General Medicine
Background: Pain is a global health issue that affects an individual's quality of life. Its allev... more Background: Pain is a global health issue that affects an individual's quality of life. Its alleviation and management will enhance patients' experience. Community pharmacists can help manage pain severity through their valuable roles in medical teams and by managing the consequences of pain. Objective: This study aimed to evaluate community pharmacists' knowledge and attitudes toward pain and pain management in Saudi Arabia. Methods: A cross-sectional study was performed to evaluate community pharmacists' knowledge and attitudes toward pain and pain management in Saudi Arabia. Pharmacists aged ≥ 21 years, with a degree in pharmacy were included in this study. Each respondent participated in an online survey covering cancer-oriented pain and assessment of pain; pharmacology; abuse of substances; and physical dependence. An independent t-test and One-way ANOVA, with least significant difference as a post-hoc test, were employed, in addition to the General Linear Regression Model using Main Effect as the model. Results: This study revealed that the pain-related knowledge and attitude among community pharmacists in Saudi Arabia were inadequate. Age (p = 0.003), work experience (p = 0.036), nature of work (p = 0.001), and work location (p = 0.003) were determined as significant factors affecting their overall knowledge and attitude toward pain. Conclusion: Overall, attempts to expand community pharmacists' knowledge and foster an appropriate attitude toward pain management among them in Saudi Arabia are highly recommended. Additional academic courses, studies, and tailored neuroscience courses will improve their awareness and knowledge of pain and pain management.
Cancer Treatment and Research Communications
International Journal Of Pharmaceutical Research And Allied Sciences
King Khalid University Journal of Health Sciences
Clinical Pharmacology: Advances and Applications
Obesity and overweight pose a threat to health and are more common than undernutrition among adul... more Obesity and overweight pose a threat to health and are more common than undernutrition among adults. It is categorized by fat accumulation and a body mass index (BMI) of > 30. A significant increase in worldwide obesity has been ongoing over several decades. Over the past few years, several strategies have been followed for weight management and to counteract the increasing prevalence of the disease; however, room for improvement with pharmacological options still exists. This review aimed to digest selected past clinical and experimental studies and understand the role of semaglutide treatment for obesity. Methods: Articles related to the clinical uses of semaglutide, mechanism of action, pharmacokinetics, pharmacodynamics, and side effects of the drug were identified. Only studies with human subjects who used Semaglutide for obesity management were included and assessed. Results: Semaglutide promotes weight loss via appetite and hunger suppression, decreases energy intake, controls eating, and depresses the relative fondness for fatty, energy-dense foods. Moreover, the relationship between obesity and Semaglutide has been widely investigated, and most studies reveal the efficacy of Semaglutide on weight loss. Overall, the pharmacokinetics of semaglutide shows a drop in glycosylated hemoglobin A1c (HbA1c) and total body weight. The usual adverse effects observed in patients treated with Semaglutide include gastrointestinal adverse events, like nausea, vomiting, diarrhea, constipation, and abdominal cramps. Conclusion: The findings from the review suggest that semaglutide appears to be beneficial, most notably in its contribution to weight reduction.
Evidence-Based Complementary and Alternative Medicine, 2022
Many medicinal plants have been utilized for centuries despite the lack of scientific evidence of... more Many medicinal plants have been utilized for centuries despite the lack of scientific evidence of their therapeutic effects. This study evaluated the phytochemical and dual biological profiling, namely, antibacterial and cytotoxic properties, of three plant species, namely, Tribulus terrestris L., Typha domingensis Pers., and Ricinus communis L., in order to explore potential relationships (if any) with their ethnopharmacological uses. GC-MS was used to achieve phytochemical screening of two plant extracts (T. terrestris and T. domingensis). The primary chemicals detected in varying amounts in both extracts were siloxane derivatives, fatty acid esters, diisooctyl phthalate, phytosterol, and aromatic acid esters. According to the findings, the major component detected in both extracts was 1,2-benzenedicarboxylic acid and diisooctyl ester (antibacterial and antifungal). T. domingensis contained a low level of benzoic acid, methyl ester (antibacterial). Both extracts included stigmaste...
Purpose: In a sample of Middle Eastern population, we sought to evaluate whether survival outcome... more Purpose: In a sample of Middle Eastern population, we sought to evaluate whether survival outcomes for mCRC patients treated with systemic chemotherapy (SC) and, with and without biologic therapies (bevacizumab and cetuximab) are different between left and right-sided tumors. Methods: Ministry of National Guard- Health Affairs (MNG-HA) Cancer registry data was utilized to identify patients diagnosed with CRC between 2013 and 2016. Kaplan-Meier method and porosity score adjusted Cox proportional hazard models were used to assess survival for right and left-sided mCRC with and with BT. Results: A total of 196 mCRC patients with mean age of 64 years old, 57.65% male diagnosed from January 1, 2013, through December 31, 2016. The overall median survival for left-sided was significantly higher than right-sided mCRC tumors (P 0.03). mCRC patients treated with SC+BT were associated with decreased mortality only among patients with left-sided mCRC compared to right-sided mCRC (HR, 0.21; 95% ...
World Journal of Pediatrics, 2022
Background Multisystem inflammatory syndrome in children (MIS-C) is a serious health condition th... more Background Multisystem inflammatory syndrome in children (MIS-C) is a serious health condition that develops from and is linked to coronavirus disease 2019. MIS-C is considered a multi-organ dysfunction involving cardiac, renal, respiratory, hematologic, gastrointestinal and neurological symptoms and groups of signs and symptoms such as rash or bilateral nonpurulent conjunctivitis, hypotension or shock and acute gastrointestinal problems, which require immediate therapeutic intervention to prevent the aggravation of the patient's health condition. MIS-C is relatively new in the field of evidence-based medicine; however, there are several clinical guidelines for good clinical practice. For every disorder, the guidelines have different suggestions. Hence, based on the current status of the evidence, recommendations have been combined to form a unified guideline for therapeutic management. Methods This paper compares and evaluates the current MIS-C-specific clinical practice guidelines (namely, American
Several studies have reported that the multifunctional enzyme tissue transglutaminase (TG2) play ... more Several studies have reported that the multifunctional enzyme tissue transglutaminase (TG2) play important roles in neurite outgrowth and modulation of neuronal cell survival. Recently, TG2 enzymatic activity has been shown to be regulated by protein kinases such as PKA and PKC, modulating through G-protein coupled receptors (GPCRs). However, the regulation of TG2 following stimulation of GPCRs coupled to PKC and PKA activation are not fully understood. In neuronal cells, the neurotrophic factors; pituitary adenylate cyclase activating polypeptide (PACAP) and nerve growth factor (NGF) promote the differentiation, maturation, neurite outgrowth and survival of neurons via their GPCR's; the pituitary adenylate cyclase type 1 receptor (PAC1) and the tyrosine kinase receptor (TrKA) including multiple protein kinase signalling pathways. As some of these kinase pathways are associated with modulation of intracellular TG2 activity, it is conceivable to hypothesised that PACAP and NGF di...
Molecules, 2021
Breast cancer is a complex and multi-drug resistant (MDR) disease, which could result in the fail... more Breast cancer is a complex and multi-drug resistant (MDR) disease, which could result in the failure of many chemotherapeutic clinical agents. Discovering effective molecules from natural products or by derivatization from known compounds is the interest of many research studies. The first objective of the present study is to investigate the cytotoxic combinatorial, chemosensitizing, and apoptotic effects of an isatin derived compound (5,5-diphenylimidazolidine-2,4-dione conjugated with 5-substituted isatin, named HAA2021 in the present study) against breast cancer cells (MCF7) and breast cancer cells resistant to doxorubicin (MCF7/ADR) when combined with doxorubicin. The second objective is to investigate the binding mode of HAA2021 withP-glycoprotein (P-gp) and heat shock protein 90 (Hsp90), and to determine whether their co-inhibition by HAA2021 contribute to the increase of the chemosensitization of MCF7/ADR cells to doxorubicin. The combination of HAA2021, at non-toxic doses, w...
Journal of Pharmaceutical Research International
Aim: In this study, we aimed to investigate the incidence rate, risk factors, and mortality rates... more Aim: In this study, we aimed to investigate the incidence rate, risk factors, and mortality rates in patients with early-stage breast cancer using anti-HER2 (Human epidermal growth factor receptor-2) treatment. Patients and Methods: A total of 106 patients diagnosed with human epidermal growth factor 2 (HER2)-positive early-stage breast cancer and receiving anti-HER2 treatment at King Abdulaziz Medical City (KAMC) from 2015 to 2019 were included in the analysis to assess the incidence of cardiotoxicity was collected as a retrospective study. Univariate and multivariate analyses as well as multiple exact logistic regression analysis were conducted to understand the relationships between the left ventricular ejection fraction (LVEF) and treatment combinations and comorbidities Results: The LVEF measurements using an echocardiography method at the baseline (before any treatment) and during the anti-HER2 therapy were assessed. The results suggest that the higher the drug combination, th...
Journal of Molecular Structure
Archives of Toxicology
Organophosphate compounds (OPs) induce both acute and delayed neurotoxic effects, the latter of w... more Organophosphate compounds (OPs) induce both acute and delayed neurotoxic effects, the latter of which is believed to involve their interaction with proteins other than acetylcholinesterase. However, few OP-binding proteins have been identified that may have a direct role in OP-induced delayed neurotoxicity. Given their ability to disrupt Ca2+ homeostasis, a key aim of the current work was to investigate the effects of sub-lethal neurite outgrowth inhibitory levels of OPs on the Ca2+-dependent enzyme tissue transglutaminase (TG2). At 1–10 µM, the OPs phenyl saligenin phosphate (PSP) and chlorpyrifos oxon (CPO) had no effect cell viability but induced concentration-dependent decreases in neurite outgrowth in differentiating N2a neuroblastoma cells. The activity of TG2 increased in cell lysates of differentiating cells exposed for 24 h to PSP and chlorpyrifos oxon CPO (10 µM), as determined by biotin-cadaverine incorporation assays. Exposure to both OPs (3 and/or 10 µM) also enhanced i...
European Journal of Pharmacology
NGF (nerve growth factor) and tissue transglutaminase (TG2) play important roles in neurite outgr... more NGF (nerve growth factor) and tissue transglutaminase (TG2) play important roles in neurite outgrowth and modulation of neuronal cell survival. In this study, we investigated the regulation of TG2 transamidase activity by NGF in retinoic acid-induced differentiating mouse N2a and human SH-SY5Y neuroblastoma cells. TG2 transamidase activity was determined using an amine incorporation and a peptide cross linking assay. In situ TG2 activity was assessed by visualising the incorporation of biotin-X-cadaverine using confocal microscopy. The role of TG2 in NGF-induced cytoprotection and neurite outgrowth was investigated by monitoring hypoxia-induced cell death and appearance of axonal-like processes, respectively. The amine incorporation and protein crosslinking activity of TG2 increased in a time and concentration-dependent manner following stimulation with NGF in N2a and SH-SY5Y cells. NGF mediated increases in TG2 activity were abolished by the TG2 inhibitors Z-DON (Z-ZON-Val-Pro-Leu-OMe; Benzyloxycarbonyl-(6-Diazo-5-oxonorleucinyl)-L-valinyl-L-prolinyl-L-leucinmethylester) and R283 (1,3,dimethyl-2[2-oxo-propyl]thio)imidazole chloride) and by pharmacological inhibition of extracellular signal-regulated kinases 1 and 2 (ERK1/2), protein kinase B (PKB) and protein kinase C (PKC), and removal of extracellular Ca 2+. Fluorescence microscopy demonstrated NGF induced in situ TG2 activity. TG2 inhibition blocked NGFinduced attenuation of hypoxia-induced cell death and neurite outgrowth in both cell lines. Together, these results demonstrate that NGF stimulates TG2 transamidase activity via a ERK1/2, PKB and PKC-dependent pathway in differentiating mouse N2a and human SH-SY5Y neuroblastoma cells. Furthermore, NGF-induced cytoprotection and neurite outgrowth are dependent upon TG2. These results suggest a novel and important role of TG2 in the cellular functions of NGF.
Biochemical Pharmacology, Mar 1, 2017
The PAC1 receptor and tissue transglutaminase (TG2) play important roles in neurite outgrowth and... more The PAC1 receptor and tissue transglutaminase (TG2) play important roles in neurite outgrowth and modulation of neuronal cell survival. In this study, we investigated the regulation of TG2 activity by the PAC1 receptor in retinoic acid-induced differentiating N2a neuroblastoma cells. TG2 transamidase activity was determined using an amine incorporation and a peptide cross linking assay. In situ TG2 activity was assessed by visualising the incorporation of biotin-X-cadaverine using confocal microscopy. TG2 phosphorylation was monitored via immunoprecipitation and Western blotting. The role of TG2 in PAC1 receptor-induced cytoprotection and neurite outgrowth was investigated by monitoring hypoxia-induced cell death and appearance of axonal-like processes, respectively. The amine incorporation and protein crosslinking activity of TG2 increased in a time and concentration-dependent manner following stimulation with pituitary adenylate cyclase-activating polypeptide-27 (PACAP-27). PACAP-27 mediated increases in TG2 activity were abolished by the TG2 inhibitors Z-DON and R283 and by pharmacological inhibition of protein kinase A (KT 5720 and Rp-cAMPs), protein kinase C (Ro 31-8220), MEK1/2 (PD 98059), and removal of extracellular Ca 2+. Fluorescence microscopy demonstrated PACAP-27 induced in situ TG2 activity. TG2 inhibition blocked PACAP-27 induced attenuation of hypoxia-induced cell death and outgrowth of axon-like processes. TG2 activation and cytoprotection were also observed in human SH-SY5Y cells. Together, these results demonstrate that TG2 activity was stimulated downstream of the PAC1 receptor via a multi protein kinase dependent pathway. Furthermore, PAC1 receptor-induced cytoprotection and neurite outgrowth are dependent upon TG2. These results highlight the importance of TG2 in the cellular functions of the PAC1 receptor.