Efect Of Qikui Granule On Rats With Diabetic Nephropathy And Expression Of MCP-1in Its Kidneys And In Human Mesangial Cells Induced By High Glucose

Objective:To observe the efect of Qikui granule on expression of MCP-1in kidneys of rats with diabetic nephropathy and human mesangial cells induced by high glucose, to explore the possible mechanisms for the treatment of diabetic nephropathy with Qikui granule.Methods:Establish a type2diabetes rat model by the high-fat diet plus intraperitoneal injection of a small dose STZ. Male SD rats were randomly divided into normal control group, model group, TCM group and irbesartan group. Body weight and blood glucose by shearing the end of trail were measured once every two weeks. The end of the8th weeks, before killed, the rats were keeped in metabolic cages to collect24h urine for urine protein (Upro), urinary albumin/creatinine (mAlb/Cr). Carotid blood was collected for determination of blood glucose (Glu), total cholesterol (TC), triglyceride (TG), blood urea nitrogen (BUN) and creatinine (Cre). After the rats were killed, taking the both kidneys, weighing kidney weight, HE staining renal tissue, observing renal changes in organizational structure and to locate and to quantitate expression of MCP-1by immunohistochemical method.The logarithmic growth phase HRMC is divided into three major groups:the crude drug group,10%of the drug-containing serum group and20%of the drug-containing serum group. The crude drug group is divided into the following six groups:normal control group (NG,5mmol/L glucose), the mannose alcohol osmotic pressure control group (Man,5mmol/L glucose+25mmol/L mannitol), high glucose group (HG,30mmol/L glucose),2ug crude drug group (S2,30mmol/L glucose+2ug/mL crude drug), lug crude drug group (S1,30mmol/L glucose+1ug/mL crude drug),0.5ug crude drug group (S0.5,30mmol/L glucose+0.5ug/mL crude drug)(crude drug concentrations were screened by MTT).10%of the drug-containing serum group was divided into the following six groups:10%normal serum low-sugar group (NLG15mmol/L glucose), high glucose group10%normal serum (NHG1,30mmol/L glucose),10%of low-dose serum low-sugar group (LLG1,5mmol/L glucose),10%of low-dose serum high glucose group (LHG1,5mmol/L glucose),10%of high-dose serum sugar group (HLG1,5mmol/L glucose),10%of high-dose serum sugar group (HHG1,5mmol/L glucose).20% of the drug-containing serum group was divided into the following six groups:20%normal serum low-sugar group (NLG2,5mmol/L glucose),20%normal serum high-sugar group (NHG2,30mmol/L glucose),20%of low-dose serum low-sugar group (LLG2,5mmol/L glucose),20%of low-dose serum high glucose group (LHG2,5mmol/L glucose),20%of the high dose of serum low-sugar group (HLG2,5mmol/L glucose),20%of high-dose serum low-sugar group (HHG2,5mmol/L glucose). Realtime-RT-PCR was used to detect the expression of24h MCP-1mRNA. The expression of MCP-1protein was detected by western blot.And MCP-1protein secreted in culture supernatant was detected by ELISA kit.Results:Compared with the model group rats weight of traditional Chinese medicine group had a significant increase (P<0.05). Kidney weight significantly reduced (P<0.05).Kidney weight/body weight ratio also significantly reduced (P<0.05). Glucose, total cholesterol, blood urea nitrogen and creatinine were significantly lower (P<0.05). The model rats glomerular hyperplasia, glomerular capillary hyalinization and sclerosis, glomerular capillary occlusion were allieviated in the traditional Chinese medicine group. And tubular interstitial MCP-1expression levels significantly reduced compared with the model group. In crude drug group, compared with HG, MCP-1protein expression in S1was significantly decreased (P<0.05). In10%serum containing group, compared with the NHG1, HHG1MCP-1mRNA expression levels decreased (P<0.05), compared with NLG1the HLG1of MCP-1protein was significantly lower (P<0.05). In20%of the drug-containing serum group, compared with NLG2, HLG2MCP-1protein expression significantly decreased (P<0.05), the HHG2of MCP-1protein levels were significantly lower (P<0.05), compared with NHG2.Conclusion:the Qikui granule can significantly lower blood glucose levels of rats with diabetic nephropathy, reduc renal MCP-1expression, anti-diabetic rat kidney disease, reduce MCP-1mRNA and protein expression of human mesangial cells induced by high glucose. Thus Qikui granule may protect the kidneys and delay the development of diabetic nephropathy kidney disease by improving kidney inflammation.