The Protection Mechanism of Huaiqihuang Granules from Diabetic Kidney Disease in Rats and for Renal Cells in High-Glucose Environment
Objective To observe the protection mechanism of Huaiqihuang Granules from diabetic kidneydisease(DKD)in rats and for renal cells in high-glucose environment.Methods(1)In vivo experiment:the rat model was constructed by high-fat diet feeding combined with STZ intraperitoneal injection.The experiment was divided into normal group,model group,Huaiqihuang Granules low-,medium-and high-dose groups and Losartan group,with 10 rats in each group.After corresponding treatment,the main biochemical indexes such as urinary microalbumin,fasting blood glucose,serum creatinine,total cholesterol and low density lipoprotein cholesterol were measured,and the pathological changes of renal tissue were observed by hematoxylin-eosin staining,periodic acid Schiff's staining and Masson staining,respectively.(2)In vitro experiments:a high glucose-induced renal tubular epithelial cell HK2 model was constructed.The experiment was divided into normal glucose group,high-glucose group,Huaiqihuang Granules group and mannitol group.After corresponding treatment,the cell proliferation was detected by MTT assay,and the generation of reactive oxygen species(ROS)in cells and mitochondria was detected by 2',7'-Dihydrodichlorofluorescein diacetate(DCFH-DA)method and MitoSOX/Hoechst 33342 staining method,respectively.Western Blot was used to detect the expressions of inflammasome-related proteins NOD-like receptor protein 3(NLRP3),cleaved-caspase-3 and interleukin-1β(IL-1β)and renal fibrosis-related proteins fibronectin(FN)and type Ⅳ collagen(Col Ⅳ).Results(1)In vivo experiments,Huaiqihuang Granules significantly improved the urinary microalbumin excretion and serum creatinine,hyperglycemia and lipid metabolism disorders in DKD rats,and reduced the pathological damage of renal tissue.(2)In vitro studies,Huaiqihuang Granules significantly inhibited proliferation and ROS overproduction of high-glucose-induced HK2 cells,reduced the activation of NLRP3 inflammasome and the expressions of renal fibrosis-related proteins FN and Col Ⅳ in high-glucose-induced HK2 cells.Conclusion Huaiqihuang Granules has an impovement effect on DKD in rats,and its protection mechanism for renal cells in high-glucose environment is relavent to inhibiting ROS generation,activation of NLRP3 inflammasome and expression of fibrosis-related proteins.
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