QDPR Gene Expression Level Influence Oxidative Stress Of NRK-52E Cells In High Glucose

Objective The contents of superoxide anion(O2-) and superoxide dismutase 1(SOD1) in cell models were detected under high glucose environment,to study whether QDPR expression level change can affect oxidative stress of NRK-52 E renal tubular cells in a high glucose environment.Methods The NRK-52 E models of overexpression QDPR, overexpression lentivirus control, knockdown QDPR gene and lentivirus SHDNA control were constructed by lentivirus in laboratory. All groups were given 5.4mmol/L and 30mmol/L culture medium respectively to imitate normal and high glucose condition. The experiment cell models are divided according to the following groups:1 NRK-52 E Control(NC)2 NRK-52 E exposed to High glucose(NHG)3 overexpression lentivirus control(LV-OCON)4 overexpression lentivirus control exposed to High glucose(LV-OCON-HG)5 lentivirus overexpressed QDPR(LV-QDPR)6 lentivirus overexpressed QDPR exposed to High glucose(LV-QDPRHG) 7 lentivirus SHDNA control(LV-SHCON) 8 lentivirus SHDNA control exposed to High glucose(LV-SHCON-HG) 9 lentivirus QDPR SHDNA(LV-SHQDPR) 10 lentivirus QDPR SHDNA exposed to High glucose(LV-SHQDPR-HG). The level of superoxide anion(O2-) was detected by flow cytometer dihydroethidium method. The protein expression level of SOD1 was tested by Western blot.Results 1 Under high glucose environment NRK-52 E cells are in the oxidative stress state. Compared with the normal glucose group(NC), O2- level increased in NHG group(P < 0.05); SOD1 expression decreased(P < 0.05). 2 Over expression QDPR gene of NRK-52 E in cell model successfully constructed by lentivirus. Under high glucose environment, the O2- content of the LV-QDPR-HG group was lower than that of the control group LVOCON-HG(P < 0.001). In high glucose environment, SOD1 expression was regulated down in the LV-QDPR-HG group(P < 0.01). 3 Knockdown QDPR gene of NRK-52 E in cell model successfully constructed by lentivirus. In high glucose environment, compared with LV-SHCON-HG, the O2- content of the LV-SHQDPR-HG group was higher(P < 0.05). Under high glucose environment, compared with LV-SHCON-HG group, the SOD1 expression had no significant change in the LV-SHQDPR-HG group(P>0.05).Conclusions 1 QDPR gene can affect the occurrence and development of diabetic nephropathy by oxidative stress. 2 Under high glucose condition, overexpression QDPR gene can decrease NRK-52 E cell oxidative stress. 3 Under high glucose condition, Knockdown QDPR gene can increase NRK-52 E cell oxidative stress.