Study On Kidney Protection Mechanism Of Shenxiao Tongluo Decoction In Diabetic Db/db Mice Based On PI3K/AKT Signal Pathway

Objective:The expression of vascular endothelial cell marker CD31 was detected,the proliferation and angiogenesis of glomerular endothelial cells in db/db mice and the intervention effect of shenxiao tongluo decoction were observed,the activation of the PI3K/AKT/VEGF signaling pathway was observed to explore the possible mechanism of shenxiao tongluo decoction,and provide experimental basis for clinical application of traditional Chinese medicineMethods:In this study,db/db mice with spontaneous type 2 diabetes were used as the DKD model.Thirtyeight-week-old male db/db mice were divided into model group?Model?,shenxiao tongluo group?SXTL?and candesartan group?KDST?,ten homology db/m mice with spontaneously type2 diabetes mellitus were divided into normal control group?Control?fed for 8 weeks.The mice of shenxiao tongluo group were intragastric administrated with Shenxiao Tongluo for 29.5 g.kg^-1.d^-11 and the candesartan group with candesartan for 1.33 mg.kg^-1.d^-1,control group and model group with distilled water of equal capacity.Body weights and blood glucose levels were measured at before and after intervention 4,8 weekends.The urine were collected to test 24h urine protein in the end.The blood was collected to measure Scr and BUN.One side of the kidney were fixed by 4%paraformaldehyde for HE and PAS staining in order to observe the renal pathological change.The expression of CD31 were detected by immunohistochemistry and semi-quantitatively analyzed,the correlation of their expression with 24h urinary protein were analyzed.The other side of kidney was kept in refrigerator at-80?.PI3K,p-PI3K,AKT,p-AKT,VEGF were detected by Western blot.Results:1.General condition of miceCompared with the control group,the mice in the model group showed obvious symptoms of polydipsia,polyphagia,polyuria and body obesity.Two died in the normal group due to intragastric administration,while two died in the model group,shenxiao tongluo group and candesartan group.2.Body weightsCompared with the control group,the model group showed higher body weights?P<0.01?.Compared with the model group,the shenxiao tongluo group and the candesartan group were decreased?P<0.05?.3.Comparison of blood glucose,renal function and 24h urinary protein contentCompared with the control group,the blood glucose of mice in the model group increased?P<0.01?.Compared with the model group,the blood glucose of mice in the shenxiao tongluo group and the candesartan group showed no significant changes.Compared with the control group,serum Scr,BUN and 24h urinary prote-in content in the model group were significantly increased?P<0.01?.Compared with the model group,the serum Scr,BUN content and 24h urinary protein in shenxiao tongluo group and candesartan group were significantly decreased?P<0.05?.4.changes of renal PathomorphologicalHE staining showed that the glomerular structure was clear and normal in the control group.The number of cells in the glomeruli increased and the glomerular hypertrophy increased in the model group.Compared with the model group,the pathological changes of shenxiao tongluo group and candesartan group were reduced,but the differences between the two groups were not significant.PAS staining showed that no obvious abnormality of glomerular structure was in the control group.The glomerular basement membrane thickened,extracellular matrix accumulated and mesangial hyperplasia occurred in the model group.Compared with the model group,the pathological changes of shenxiao tongluo group and candesartan group were reduced.5.The expression of CD31 were examined by immunohistochemistryCompared with the control group,the expression of glomerular CD31 in the model group was significantly increased?P<0.01?.Compared with the model group,the expressions of CD31 in the glomerular glomeruli of shenxiao tongluo group and candesartan group were significantly decreased?P<0.05?.6.Correlation analysis of CD31 expression with 24h urinary proteinCorrelation analysis showed that CD31 expression was positively correlated with 24h urinary protein.7.The expression of PI3K,p-PI3K,AKT,p-AKT,VEGF were detected by Western blotWestern blot results showed that the expression of p-PI3K/PI3K,p-AKT/AKT and VEGF in the model group was significantly enhanced compared with the control group?P<0.01?.After treatment,the expression of p-PI3K/PI3K,p-AKT/AKT and VEGF in the shenxiao tongluo group and candesartan group was significantly decreased compared with the model group,and the difference was statistically significant.Conclusion:1.Shenxiao tongluo could improve the general condition of db/db mice,reduce the content of urine protein for 24h and the level of Scr,BUN,improve the renal function.2.Shenxiao tongluo can alleviate the pathological damage of kidney in db/db mice.3.Shenxiao tongluo can suppress the high expression of CD31.4.Shenxiao tongluo can suppress the PI3K/AKT/VEGF signal pathway and the proliferation of endothelial cells and the generation of blood vessels,which may be one of the mechanisms for its treatment of DKD.