ABSTRACT
<p><b>AIM</b>To investigate the effect and mechanism of ascorbic acid on podocyte, last barrier of glomerular filtration, in diabetic rats.</p><p><b>METHODS</b>Diabetic rats induced by streptozotocin injection intraperitoneally were treated by ascorbic acid for 5 weeks. The levels of blood glucose (BG), HbA1c, urinary albumin excretion rate (UAER) and superoxide diamutase (SOD), catalase (CAT) and malondialdehyde (MDA) in renal cortex were measured. The podocyte ultrastructure was observed while the expression of desmin protein, a marker of podocyte injury, was examined.</p><p><b>RESULTS</b>Compared with control group, BG and HbA1c were increased markedly in diabetic group. The activities of SOD and CAT were decreased and the concentrations of MDA were increased significantly in diabetic renal cortex. There were the increased proteinic expression of desmin, foot process effacement in podocytes and UAER markedly in diabetic rats. Compared with diabetic rats, foot process effacement and the changes of UAER were ameliorated markedly while the activities of SOD were increased, the levels of MDA and proteinic expression of desmin were decreased markedly although BG, HbA1c and the activities of CAT were no significant difference in the diabetic rats by ascorbic acid treatment.</p><p><b>CONCLUSION</b>The findings suggest that there are marked injury in podocyte, last barrier of glomerular filtration, in diabetic rats and administration of ascorbic acid can protect podocyte by increasing antioxidative capacity and ameliorating the renal oxidative stress.</p>
Subject(s)
Animals , Male , Rats , Ascorbic Acid , Pharmacology , Catalase , Metabolism , Desmin , Metabolism , Diabetes Mellitus, Experimental , Metabolism , Pathology , Diabetic Nephropathies , Metabolism , Pathology , Oxidative Stress , Podocytes , Metabolism , Random Allocation , Rats, Wistar , Superoxide Dismutase , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effects of recombinant adenovirus encoding human apM1 gene on proliferation and nitric oxide synthase (NOS) activity in human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>Protein expression of apM1 in cell culture supernatant of HUVECs transfected with human Ad-apM1 was detected by double antibody sandwich ELISA. The effect of human adiponectin on cell proliferation was assessed by MTT assay. The total NOS and iNOS expressions were measured by chromatometre.</p><p><b>RESULTS</b>Human adiponectin protein level and total NOS and eNOS expressions were significant increased and iNOS expression significantly reduced in culture supernatant of HUVECs infected with Ad-apM1 compared to that in control HUVECs. The recombinant adenovirus had no influence on HUVECs growth as determined by MTT assay.</p><p><b>CONCLUSIONS</b>Human Ad-apM1 can be effectively expressed in HUVECs and do not influence HUVECs growth. Increased total NOS and eNOS expressions and decreased iNOS expression in HUVECs transfected with Ad-apM1 gene suggest a potential role of Ad-apM1 gene transfer for the prevention and treatment of arteriosclerosis.</p>
Subject(s)
Humans , Adenoviridae , Genetics , Adiponectin , Genetics , Cell Proliferation , Cells, Cultured , Endothelial Cells , Cell Biology , Metabolism , Endothelium, Vascular , Cell Biology , Gene Expression , Gene Transfer Techniques , Nitric Oxide Synthase , Metabolism , Umbilical Veins , Cell BiologyABSTRACT
Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.