Effect Of Keluoxin Capsule On Renal Function And Podocyte Autophagy In Rat Models Of Diabetic Nephropathy

Objective: To observe the effect of Keluoxin capsule on renal function and autophagy of podocytes in diabetic nephropathy rat models induced by STZ combined with high fat and high sugar diet.Methods: 40 SPF grade male SD rats,10 were randomly selected as control group,and the remaining 30 rats were induced diabetic nephropathy rats model by intraperitoneal streptozotocin(STZ)and high fat and high glucose diet.After successful modeling,they were randomly divided into DN group(model group),Keluoxin group and Irbesartan group,with 10 rats in each group.After 8 weeks of treatment,the body weight was measured and 24 h urine volume was recorded,the levels of FBG,Urea,MAU and MAU/UCr were detected,and the renal tissues were stained by PAS and Masson staining to observe the pathological changes of the kidneys.Immunofluorescence staining was used to observe the expression of Nephrin protein in podocytes,and Western Blot was used to detect the expression of autophagy-related proteins in podocytes.Results:(1)In the model group,the body weight of model rats decreased obviously(P<0.01),and the levels of FBG,24 h urinary volume,Urea,MAU and MAU/UCr increased significantly(P<0.01).Compared with the model group,the body weight,FBG and MAU/UCR levels in Keluoxin group decreased significantly(P<0.01),and the 24 h urine volume,Urea and MAU levels decreased obviously(P<0.05).In Irbesartan group,the 24 h urine volume decreased obviously(P<0.05),the body weight,MAU and MAU/UCR levels decreased significantly(P<0.01),while there are no significant improvement in FBG and Urea levels.(2)PAS staining: In the model group,the rat glomerular basement membrane became irregularly thick,the mesangium substrate increased,and the percentage of glomerular positive expression areas increased clearly(P<0.01).After treatment with Keluoxin and Irbesartan,the percentage of glomerular positive expression areas decreased significantly(P<0.01).(3)Masson staining: In the model group,a large number of blue areas appeared in the kidney tissues,and the percentage of collagen expression areas increased significantly(P<0.01).After treatment with Keluoxin and Irbesartan,the blue areas of renal tissues and the percentage of collagen expression areas decreased significantly(P<0.05,P<0.01).(4)Immunofluorescence detection: Nephrin protein content in model group decreased significantly from the control group,and the mean fluorescence intensity was significantly lowered(P<0.05).After treatment with Keluoxin and Irbesartan,the Nephrin protein content and the mean fluorescence intensity of the two groups increased significantly(P<0.05).(5)Western blot test: the results showed that the expression of Nephrin in the model group decreased significantly,while the levels of LC3-ll,LC3-II/LC3-I and P62 increased significantly(P<0.01).After treatment with Keluoxin and Irbesartan,the Nephrin expression in Keluoxin group was significantly up-regulated,the LC3-II and P62 levels were significantly decreased(P<0.01),and the level of LC3-II/LC3-I reduced significantly(P<0.05).In Irbesartan group,the expression of Nephrin was significantly up-regulated,the LC3-II and P62 levels decreased significantly(P<0.01).Conclusion: The Keluoxin capsule could reduce the body weight,24 h urine volume,FBG,Urea,MAU and MAU/UCr levels of DN rat models,improve renal function,alleviate renal histopathological changes,improve the autophagy activity of podocytes,and reduce the damage of podocytes.