12 results on '"Shenqiang, Rao"'
Search Results
2. Protective Effects of Oxymatrine on Vascular Endothelial Cells from High-Glucose-Induced Cytotoxicity by Inhibiting the Expression of A2B Receptor
- Author
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Yun Yi, Yulin Shen, Qin Wu, Jingan Rao, Shu Guan, Shenqiang Rao, Liping Huang, Mengxia Tan, Lingkun He, Lijuan Liu, Guodong Li, Shangdong Liang, Wei Xiong, and Yun Gao
- Subjects
A2B receptor ,Oxymatrine ,Human umbilical vein ,Endothelial cells ,Diabetes mellitus ,Physiology ,QP1-981 ,Biochemistry ,QD415-436 - Abstract
Background/Aims: Diabetes mellitus (DM) has become an increasingly epidemic metabolic disease. Vascular endothelial cells play a key role in developing the cardiovascular complications of DM. The A2B receptor is expressed in vascular endothelial cells, and may help regulate the function of endothelial cells. The aim of this study was to investigate the protective effects of oxymatrine (OMT) on human umbilical vein endothelial cells (HUVECs) from high glucose-induced cytotoxicity. Methods: Homology modeling and molecular docking analysis were used to detect the binding sites between the adenosine A2B receptor and OMT. HUVECs were cultured with control (5.5 mM) or elevated glucose (22.2 mM) in the presence or absence of 3 µM OMT or A2B siRNA for 3 days. The MTS cell viability assay was used to measure the toxicity of high glucose on HUVECs and the protective effect of OMT or A2B siRNA. The expression of the adenosine A2B receptor and CCL5 in HUVECs was detected with real-time quantitative PCR (qPCR) and Western blotting methods in each group. Levels of IL-1β and TNF-α were measured using an enzyme-linked immunosorbent assay (ELISA) kit, and the concentration of NO was detected with the nitrate reductase method. Monocyte chemotactic activity in each group was detected using Transwell chambers. Furthermore, the phosphorylation of p38 and ERK1/2 in each group was observed through the Western blotting method. Results: Homology modeling and molecular docking analysis showed that OMT contains well-fitted binding sites to the A2B receptor. After chronic culture at high glucose, the rate of cell viability was significantly lower than that of the control group. After co-treatment with OMT or A2B siRNA, cell viability was significantly increased compared with the high-glucose group. The results from real-time quantitative RT-PCR (qRT-PCR) and Western blotting indicated that high glucose could increase the expression of A2B receptors in HUVECs, an effect that was inhibited by OMT. In addition, the results revealed that the expression of CCL5, IL-1β and TNF-α was increased in the high-glucose group, and that the NO produced by HUVECs decreased due to hyperglycemia; however, co-culture with OMT or A2B siRNA abolished these effects. Meanwhile, the chemotaxis activity of monocytes to HUVECs cultured in high-glucose medium was enhanced 2.59-fold compared to the control cells. However, the inflammatory reactions in HUVECs were completely relieved by co-treatment with OMT or A2B siRNA. Moreover, the phosphorylation of p38 and ERK1/2 in HUVECs in the high-glucose group was significantly higher than that of the control group; these effects were reversed after co-treatment with OMT or A2B siRNA. Conclusion: OMT may protect the HUVECs from high glucose-induced cytotoxicity through inhibitting the expression of A2B receptor and inflammatory factors as well as decreasing the phosphorylation of p38 and ERK1/2.
- Published
- 2018
- Full Text
- View/download PDF
3. Naringin Protects Against High Glucose-Induced Human Endothelial Cell Injury Via Antioxidation and CX3CL1 Downregulation
- Author
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Guilin Li, Yurong Xu, Xuan Sheng, Hua Liu, Jingjing Guo, Jiayue Wang, Qi Zhong, Huaide Jiang, Chaoran Zheng, Mengxia Tan, Shenqiang Rao, Yanling Yu, Yun Gao, Guodong Li, Shangdong Liang, and Gaochun Zhu
- Subjects
Naringin ,HUVEC ,CX3CL1 ,ROS ,Mitochondria ,Oxygen consumption rate ,Physiology ,QP1-981 ,Biochemistry ,QD415-436 - Abstract
Background/Aims: The induction of endothelial injury by hyperglycemia in diabetes has been widely accepted. Naringin is a bio-flavonoid. Some studies showed that naringin alleviates diabetic complications, but the exact mechanisms by which naringin improves diabetic anomalies are not yet fully understood. The aim of this research was to study the protective effect of naringin on high glucose-induced injury of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were cultured with or without high glucose in the absence or presence of naringin for 5 days. The expression of CX3CL1 was determined by quantitative real-time RT-PCR (qPCR) and western blot. The cellular bioenergetic analysis oxygen consumption rate (OCR) was measured with a Seahorse Bioscience XF analyzer. Results: The production of reactive oxygen species (ROS), the expression of CX3CL1 and the level of AKT phosphorylation were increased in HUVECs cultured with high glucose compared with controls. However, naringin rescued these increases in ROS production, CX3CL1 expression and AKT phosphorylation. Nitric oxide (NO) production and OCR were lower in the high glucose group, and naringin restored the changes induced by high glucose. Molecular docking results suggested that Naringin might interact with the CX3CL1 protein. Conclusion: Naringin protects HUVECs from high-glucose-induced damage through its antioxidant properties by downregulating CX3CL1 and by improving mitochondrial function.
- Published
- 2017
- Full Text
- View/download PDF
4. Purinergic P2X7 receptor functional genetic polymorphisms are associated with the susceptibility to osteoporosis in Chinese postmenopausal women
- Author
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Luling He, Yongfang Fan, Fangfang Hu, Peipei Zhong, Yuping Yang, Tao Li, Shangdong Liang, Changle Liu, Xingzi Liu, Shuo Wang, Lan Tang, Hong Xu, Shenqiang Rao, Lu Ding, Chaopeng Xiong, Yijun Nie, Yunming Tu, Chengxin Gong, and Hui Wang
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Candidate gene ,Genotype ,Osteoporosis ,Single-nucleotide polymorphism ,Biology ,Polymorphism, Single Nucleotide ,Pathogenesis ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Sex Factors ,Asian People ,Gene Frequency ,Osteoclast ,Internal medicine ,medicine ,Humans ,Genetic Predisposition to Disease ,Allele ,Molecular Biology ,Haplotype ,Cell Biology ,medicine.disease ,Postmenopause ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Case-Control Studies ,Immunology ,Original Article ,Female ,Receptors, Purinergic P2X7 - Abstract
Osteoporosis (OP) is a major public health problem worldwide. Genetic factors are considered to be major contributors to the pathogenesis of OP. The purinergic P2X7 receptor (P2X7R) has been shown to play a role in the regulation of osteoblast and osteoclast activity and has been considered as an important candidate gene for OP. A case-control study was performed to investigate the associations of functional single nucleotide polymorphisms (SNPs) in the P2X7R gene (rs2393799, rs7958311, rs1718119, rs2230911, and rs3751143) with susceptibility to OP in 400 Chinese OP patients and 400 controls. Results showed that rs3751143 was associated with OP; in particular, carriers of the C allele and CC/(AC + CC) genotypes were at a higher risk of OP, but no significant association of rs2230911, rs7958311, rs1718119, and rs2393799 with OP risk was observed. Analysis of the haplotypes revealed one haplotype (rs1718119G-rs2230911G-rs3751143C) that appeared to be a significant "risk" haplotype with OP. The rs3751143 polymorphism was associated with osteoclast apoptosis; ATP-induced caspase-1 activity of osteoclasts with AC and CC genotypes is lower than that of osteoclasts with AA genotype in vitro. The findings suggest that the P2X7R rs3751143 functional polymorphism might contribute to OP susceptibility in Chinese postmenopausal women.
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- 2017
5. The effect of sinomenine in diabetic neuropathic pain mediated by the P2X3 receptor in dorsal root ganglia
- Author
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Shenqiang Rao, Shuangmei Liu, Lifang Zou, Tianyu Jia, Shanhong Zhao, Bing Wu, Zhihua Yi, Shouyu Wang, Yun Xue, Yun Gao, Changshui Xu, Guilin Li, Hong Xu, Chunping Zhang, and Shangdong Liang
- Subjects
Male ,Pain Threshold ,0301 basic medicine ,Purinergic P2X Receptor Antagonists ,endocrine system diseases ,Diabetes Mellitus, Experimental ,Rats, Sprague-Dawley ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Diabetic Neuropathies ,Ganglia, Spinal ,Animals ,Humans ,Molecular Biology ,nutritional and metabolic diseases ,Cell Biology ,Rats ,body regions ,HEK293 Cells ,030104 developmental biology ,Diabetes Mellitus, Type 2 ,Morphinans ,Hyperalgesia ,Original Article ,Erratum ,Receptors, Purinergic P2X3 ,030217 neurology & neurosurgery - Abstract
Type 2 diabetes mellitus (T2DM) accounts for more than 90% of all cases of diabetes mellitus (DM). Diabetic neuropathic pain (DNP) is a common complication of T2DM. Sinomenine is a natural bioactive component extracted from the Sinomenium acutum and has anti-inflammatory effects. The aim of our study was to investigate the effects of sinomenine on DNP mediated by the P2X3 receptor in dorsal root ganglia (DRG). The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) in T2DM rats were lower than those of control rats. MWT and TWL in T2DM rats treated with sinomenine were higher compared with those in T2DM rats. The expression levels of the P2X3 protein and mRNA in T2DM rat DRG were higher compared with those of the control, while those in T2DM rats treated with sinomenine were significantly lower compared with those of the T2DM rats. Sinomenine significantly inhibited P2X3 agonist ATP-activated currents in HEK293 cells transfected with the P2X3 receptor. Sinomenine decreased the phosphorylation and activation of P38MAPK in T2DM DRG. Therefore, sinomenine treatment may suppress the up-regulated expression and activation of the P2X3 receptor and relieve the hyperalgesia potentiated by the activation of P38MAPK in T2DM rats.
- Published
- 2017
6. Naringin Protects Against High Glucose-Induced Human Endothelial Cell Injury Via Antioxidation and CX3CL1 Downregulation
- Author
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Yun Gao, Guilin Li, Jiayue Wang, Guodong Li, Yurong Xu, Chaoran Zheng, Yanling Yu, Hua Liu, Shenqiang Rao, Gaochun Zhu, Jingjing Guo, Shangdong Liang, Mengxia Tan, Xuan Sheng, Huaide Jiang, and Qi Zhong
- Subjects
0301 basic medicine ,Cell Survival ,Physiology ,Down-Regulation ,Oxygen consumption rate ,Pharmacology ,Mitochondrion ,Nitric Oxide ,Protective Agents ,lcsh:Physiology ,Umbilical vein ,CX3CL1 ,Nitric oxide ,lcsh:Biochemistry ,03 medical and health sciences ,chemistry.chemical_compound ,HUVEC ,Oxygen Consumption ,0302 clinical medicine ,Downregulation and upregulation ,Western blot ,Catalytic Domain ,Human Umbilical Vein Endothelial Cells ,medicine ,Humans ,lcsh:QD415-436 ,Phosphorylation ,Naringin ,Cell Proliferation ,chemistry.chemical_classification ,Reactive oxygen species ,Binding Sites ,lcsh:QP1-981 ,medicine.diagnostic_test ,Chemokine CX3CL1 ,ROS ,Mitochondria ,Molecular Docking Simulation ,Endothelial stem cell ,Oxidative Stress ,Glucose ,030104 developmental biology ,chemistry ,Biochemistry ,030220 oncology & carcinogenesis ,Flavanones ,Reactive Oxygen Species ,Proto-Oncogene Proteins c-akt - Abstract
Background/Aims: The induction of endothelial injury by hyperglycemia in diabetes has been widely accepted. Naringin is a bio-flavonoid. Some studies showed that naringin alleviates diabetic complications, but the exact mechanisms by which naringin improves diabetic anomalies are not yet fully understood. The aim of this research was to study the protective effect of naringin on high glucose-induced injury of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were cultured with or without high glucose in the absence or presence of naringin for 5 days. The expression of CX3CL1 was determined by quantitative real-time RT-PCR (qPCR) and western blot. The cellular bioenergetic analysis oxygen consumption rate (OCR) was measured with a Seahorse Bioscience XF analyzer. Results: The production of reactive oxygen species (ROS), the expression of CX3CL1 and the level of AKT phosphorylation were increased in HUVECs cultured with high glucose compared with controls. However, naringin rescued these increases in ROS production, CX3CL1 expression and AKT phosphorylation. Nitric oxide (NO) production and OCR were lower in the high glucose group, and naringin restored the changes induced by high glucose. Molecular docking results suggested that Naringin might interact with the CX3CL1 protein. Conclusion: Naringin protects HUVECs from high-glucose-induced damage through its antioxidant properties by downregulating CX3CL1 and by improving mitochondrial function.
- Published
- 2017
7. Protective Effects of Oxymatrine on Vascular Endothelial Cells from High-Glucose-Induced Cytotoxicity by Inhibiting the Expression of A2B Receptor
- Author
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Liping Huang, Qin Wu, Yun Yi, Shangdong Liang, Wei Xiong, Jingan Rao, Mengxia Tan, Yun Gao, Shu Guan, Shenqiang Rao, Lingkun He, Lijuan Liu, Guodong Li, and Yulin Shen
- Subjects
0301 basic medicine ,Physiology ,Cell Survival ,Endothelial cells ,Interleukin-1beta ,Gene Expression ,Oxymatrine ,Protective Agents ,Receptor, Adenosine A2B ,p38 Mitogen-Activated Protein Kinases ,lcsh:Physiology ,Umbilical vein ,Monocytes ,A2B receptor ,lcsh:Biochemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Diabetes mellitus ,Alkaloids ,Human umbilical vein ,medicine ,Human Umbilical Vein Endothelial Cells ,Humans ,lcsh:QD415-436 ,Viability assay ,Phosphorylation ,Cytotoxicity ,Receptor ,Chemokine CCL5 ,Mitogen-Activated Protein Kinase 1 ,Mitogen-Activated Protein Kinase 3 ,lcsh:QP1-981 ,Tumor Necrosis Factor-alpha ,Monocyte ,Molecular biology ,Protein Structure, Tertiary ,Blot ,Molecular Docking Simulation ,030104 developmental biology ,medicine.anatomical_structure ,Glucose ,chemistry ,RNA Interference ,Adenosine A2B receptor ,Quinolizines ,Signal Transduction - Abstract
Background/Aims: Diabetes mellitus (DM) has become an increasingly epidemic metabolic disease. Vascular endothelial cells play a key role in developing the cardiovascular complications of DM. The A2B receptor is expressed in vascular endothelial cells, and may help regulate the function of endothelial cells. The aim of this study was to investigate the protective effects of oxymatrine (OMT) on human umbilical vein endothelial cells (HUVECs) from high glucose-induced cytotoxicity. Methods: Homology modeling and molecular docking analysis were used to detect the binding sites between the adenosine A2B receptor and OMT. HUVECs were cultured with control (5.5 mM) or elevated glucose (22.2 mM) in the presence or absence of 3 µM OMT or A2B siRNA for 3 days. The MTS cell viability assay was used to measure the toxicity of high glucose on HUVECs and the protective effect of OMT or A2B siRNA. The expression of the adenosine A2B receptor and CCL5 in HUVECs was detected with real-time quantitative PCR (qPCR) and Western blotting methods in each group. Levels of IL-1β and TNF-α were measured using an enzyme-linked immunosorbent assay (ELISA) kit, and the concentration of NO was detected with the nitrate reductase method. Monocyte chemotactic activity in each group was detected using Transwell chambers. Furthermore, the phosphorylation of p38 and ERK1/2 in each group was observed through the Western blotting method. Results: Homology modeling and molecular docking analysis showed that OMT contains well-fitted binding sites to the A2B receptor. After chronic culture at high glucose, the rate of cell viability was significantly lower than that of the control group. After co-treatment with OMT or A2B siRNA, cell viability was significantly increased compared with the high-glucose group. The results from real-time quantitative RT-PCR (qRT-PCR) and Western blotting indicated that high glucose could increase the expression of A2B receptors in HUVECs, an effect that was inhibited by OMT. In addition, the results revealed that the expression of CCL5, IL-1β and TNF-α was increased in the high-glucose group, and that the NO produced by HUVECs decreased due to hyperglycemia; however, co-culture with OMT or A2B siRNA abolished these effects. Meanwhile, the chemotaxis activity of monocytes to HUVECs cultured in high-glucose medium was enhanced 2.59-fold compared to the control cells. However, the inflammatory reactions in HUVECs were completely relieved by co-treatment with OMT or A2B siRNA. Moreover, the phosphorylation of p38 and ERK1/2 in HUVECs in the high-glucose group was significantly higher than that of the control group; these effects were reversed after co-treatment with OMT or A2B siRNA. Conclusion: OMT may protect the HUVECs from high glucose-induced cytotoxicity through inhibitting the expression of A2B receptor and inflammatory factors as well as decreasing the phosphorylation of p38 and ERK1/2.
- Published
- 2017
8. Resveratrol-decreased hyperalgesia mediated by the P2X
- Author
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Bing, Wu, Yucheng, Ma, Zhihua, Yi, Shuangmei, Liu, Shenqiang, Rao, Lifang, Zou, Shouyu, Wang, Yun, Xue, Tianyu, Jia, Shanhong, Zhao, Liran, Shi, Lin, Li, Huilong, Yuan, and Shangdong, Liang
- Subjects
Male ,viruses ,Blotting, Western ,dorsal root ganglia ,HIV Envelope Protein gp120 ,resveratrol ,Real-Time Polymerase Chain Reaction ,Interleukin-10 ,Rats ,Electrophysiology ,Rats, Sprague-Dawley ,HEK293 Cells ,Hyperalgesia ,HIV gp120-associated neuropathic pain ,Stilbenes ,P2X7 receptor ,Animals ,Humans ,Neuralgia ,Receptors, Purinergic P2X7 ,Research Article - Abstract
Background Chronic pain is a common symptom in human immunodeficiency virus (HIV)-1 infection/acquired immunodeficiency syndrome patients. The literature shows that the HIV envelope glycoprotein 120 (gp120) can directly cause hyperalgesia by stimulating primary sensory afferent nerves. The P2X7 receptor in the dorsal root ganglia (DRG) is closely related to neuropathic and inflammatory pain. In this study, we aimed to explore the effect of resveratrol (RES) on gp120-induced neuropathic pain that is mediated by the P2X7 receptor in the rat DRG. Results Mechanical hyperalgesia in rats treated with gp120 was increased compared with that in the sham group. The P2X7 expression levels in rats treated with gp120 were higher than those in the sham group. Co-localization of the P2X7 receptor and glial fibrillary acidic protein (GFAP, a marker of satellite glial cells [SGCs]) in the DRG SGCs of the gp120 group exhibited more intense staining than that of the sham group. RES decreased the mechanical hyperalgesia and P2X7 expression levels in gp120 treatment rats. Co-localization of the P2X7 receptor and GFAP in the gp120+ RES group was significantly decreased compared to the gp120 group. RES decreased the IL-1β and TNF-α receptor (R) expression levels and ERK1/2 phosphorylation levels as well as increased IL-10 expression in the DRG of gp120-treated rats. Whole cell clamping demonstrated that RES significantly inhibited adenosine triphosphate-activated currents in HEK293 cells that were transfected with the P2X7 plasmid. Conclusions RES relieved mechanical hyperalgesia in gp120-treated rats by inhibiting the P2X7 receptor.
- Published
- 2017
9. Erratum to: The effect of sinomenine in diabetic neuropathic pain mediated by the P2X3 receptor in dorsal root ganglia
- Author
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Bing Wu, Changshui Xu, Shouyu Wang, Tianyu Jia, Chunping Zhang, Hong Xu, Yun Xue, Shanhong Zhao, Lifang Zou, Shuangmei Liu, Guilin Li, Shangdong Liang, Shenqiang Rao, Zhihua Yi, and Yun Gao
- Subjects
0301 basic medicine ,Agonist ,food.ingredient ,endocrine system diseases ,medicine.drug_class ,Pharmacology ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,food ,Diabetes mellitus ,Threshold of pain ,Medicine ,Molecular Biology ,Sinomenine ,business.industry ,nutritional and metabolic diseases ,Type 2 Diabetes Mellitus ,Cell Biology ,medicine.disease ,body regions ,030104 developmental biology ,Anesthesia ,Hyperalgesia ,Neuropathic pain ,medicine.symptom ,business ,030217 neurology & neurosurgery ,Sinomenium - Abstract
Type 2 diabetes mellitus (T2DM) accounts for more than 90% of all cases of diabetes mellitus (DM). Diabetic neuropathic pain (DNP) is a common complication of T2DM. Sinomenine is a natural bioactive component extracted from the Sinomenium acutum and has anti-inflammatory effects. The aim of our study was to investigate the effects of sinomenine on DNP mediated by the P2X3 receptor in dorsal root ganglia (DRG). The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) in T2DM rats were lower than those of control rats. MWT and TWL in T2DM rats treated with sinomenine were higher compared with those in T2DM rats. The expression levels of the P2X3 protein and mRNA in T2DM rat DRG were higher compared with those of the control, while those in T2DM rats treated with sinomenine were significantly lower compared with those of the T2DM rats. Sinomenine significantly inhibited P2X3 agonist ATP-activated currents in HEK293 cells transfected with the P2X3 receptor. Sinomenine decreased the phosphorylation and activation of P38MAPK in T2DM DRG. Therefore, sinomenine treatment may suppress the up-regulated expression and activation of the P2X3 receptor and relieve the hyperalgesia potentiated by the activation of P38MAPK in T2DM rats.
- Published
- 2017
10. Resveratrol-decreased hyperalgesia mediated by the P2X7 receptor in gp120-treated rats
- Author
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Shenqiang Rao, Lifang Zou, Yun Xue, Shangdong Liang, Shanhong Zhao, Tianyu Jia, Lin Li, Liran Shi, Yucheng Ma, Bing Wu, Shouyu Wang, Huilong Yuan, Shuangmei Liu, and Zhihua Yi
- Subjects
0301 basic medicine ,Human immunodeficiency virus (HIV) ,Resveratrol ,medicine.disease_cause ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,0302 clinical medicine ,Acquired immunodeficiency syndrome (AIDS) ,medicine ,P2x7 receptor ,chemistry.chemical_classification ,business.industry ,virus diseases ,medicine.disease ,030104 developmental biology ,Anesthesiology and Pain Medicine ,chemistry ,Anesthesia ,Immunology ,Hyperalgesia ,Molecular Medicine ,medicine.symptom ,business ,Glycoprotein ,Hiv envelope ,030217 neurology & neurosurgery - Abstract
BackgroundChronic pain is a common symptom in human immunodeficiency virus (HIV)-1 infection/acquired immunodeficiency syndrome patients. The literature shows that the HIV envelope glycoprotein 120...
- Published
- 2017
11. Protective Effects of Oxymatrine on Vascular Endothelial Cells from High-Glucose-Induced Cytotoxicity by Inhibiting the Expression of A2B Receptor.
- Author
-
Yun Yi, Yulin Shen, Qin Wu, Jingan Rao, Shu Guan, Shenqiang Rao, Liping Huang, Mengxia Tan, Lingkun He, Lijuan Liu, Guodong Li, Shangdong Liang, Wei Xiong, and Yun Gao
- Subjects
VASCULAR endothelial cells -- Viability ,CELL-mediated cytotoxicity ,THERAPEUTIC use of alkaloids ,ADENOSINES ,PHYSIOLOGICAL effects of glucose ,DIABETES complications ,MOLECULAR docking ,PHOSPHORYLATION - Abstract
Background/Aims: Diabetes mellitus (DM) has become an increasingly epidemic metabolic disease. Vascular endothelial cells play a key role in developing the cardiovascular complications of DM. The A
2B receptor is expressed in vascular endothelial cells, and may help regulate the function of endothelial cells. The aim of this study was to investigate the protective effects of oxymatrine (OMT) on human umbilical vein endothelial cells (HUVECs) from high glucoseinduced cytotoxicity. Methods: Homology modeling and molecular docking analysis were used to detect the binding sites between the adenosine A2B receptor and OMT. HUVECs were cultured with control (5.5 mM) or elevated glucose (22.2 mM) in the presence or absence of 3 µM OMT or A2B siRNA for 3 days. The MTS cell viability assay was used to measure the toxicity of high glucose on HUVECs and the protective effect of OMT or A2B siRNA. The expression of the adenosine A2B receptor and CCL5 in HUVECs was detected with real-time quantitative PCR (qPCR) and Western blotting methods in each group. Levels of IL-1ß and TNF-a were measured using an enzyme-linked immunosorbent assay (ELISA) kit, and the concentration of NO was detected with the nitrate reductase method. Monocyte chemotactic activity in each group was detected using Transwell chambers. Furthermore, the phosphorylation of p38 and ERK1/2 in each group was observed through the Western blotting method. Results: Homology modeling and molecular docking analysis showed that OMT contains well-fitted binding sites to the A2B receptor. After chronic culture at high glucose, the rate of cell viability was significantly lower than that of the control group. After co-treatment with OMT or A2B siRNA, cell viability was significantly increased compared with the high-glucose group. The results from real-time quantitative RT-PCR (qRT-PCR) and Western blotting indicated that high glucose could increase the expression of A2B receptors in HUVECs, an effect that was inhibited by OMT. In addition, the results revealed that the expression of CCL5, IL-1ß and TNF-a was increased in the high-glucose group, and that the NO produced by HUVECs decreased due to hyperglycemia; however, co-culture with OMT or A2B siRNA abolished these effects. Meanwhile, the chemotaxis activity of monocytes to HUVECs cultured in high-glucose medium was enhanced 2.59-fold compared to the control cells. However, the inflammatory reactions in HUVECs were completely relieved by co-treatment with OMT or A2B siRNA. Moreover, the phosphorylation of p38 and ERK1/2 in HUVECs in the high-glucose group was significantly higher than that of the control group; these effects were reversed after co-treatment with OMT or A2B siRNA. Conclusion: OMT may protect the HUVECs from high glucose-induced cytotoxicity through inhibitting the expression of A2B receptor and inflammatory factors as well as decreasing the phosphorylation of p38 and ERK1/2. [ABSTRACT FROM AUTHOR]- Published
- 2018
- Full Text
- View/download PDF
12. Resveratrol-decreased hyperalgesia mediated by the P2X7 receptor in gp120-treated rats.
- Author
-
Bing Wu, Yucheng Ma, Zhihua Yi, Shuangmei Liu, Shenqiang Rao, Lifang Zou, Shouyu Wang, Yun Xue, Tianyu Jia, Shanhong Zhao, Liran Shi, Lin Li, Huilong Yuan, and Shangdong Liang
- Subjects
HIV ,RESVERATROL ,DORSAL root ganglia ,CHARCOT joints ,PELVIC inflammatory disease - Abstract
Background: Chronic pain is a common symptom in human immunodeficiency virus (HIV)-1 infection/acquired immunodeficiency syndrome patients. The literature shows that the HIV envelope glycoprotein 120 (gp120) can directly cause hyperalgesia by stimulating primary sensory afferent nerves. The P2X
7 receptor in the dorsal root ganglia (DRG) is closely related to neuropathic and inflammatory pain. In this study, we aimed to explore the effect of resveratrol (RES) on gp120-induced neuropathic pain that is mediated by the P2X7 receptor in the rat DRG. Results: Mechanical hyperalgesia in rats treated with gp120 was increased compared with that in the sham group. The P2X7 expression levels in rats treated with gp120 were higher than those in the sham group. Co-localization of the P2X7 receptor and glial fibrillary acidic protein (GFAP, a marker of satellite glial cells [SGCs]) in the DRG SGCs of the gp120 group exhibited more intense staining than that of the sham group. RES decreased the mechanical hyperalgesia and P2X7 expression levels in gp120 treatment rats. Co-localization of the P2X7 receptor and GFAP in the gp120þ RES group was significantly decreased compared to the gp120 group. RES decreased the IL-1b and TNF-a receptor (R) expression levels and ERK1/2 phosphorylation levels as well as increased IL-10 expression in the DRG of gp120-treated rats. Whole cell clamping demonstrated that RES significantly inhibited adenosine triphosphate-activated currents in HEK293 cells that were transfected with the P2X7 plasmid. Conclusions: RES relieved mechanical hyperalgesia in gp120-treated rats by inhibiting the P2X7 receptor. [ABSTRACT FROM AUTHOR]- Published
- 2017
- Full Text
- View/download PDF
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