The Effect Of Qinghua Gushen Paidu Decoction In UUO Model Of TGF-?1 And Notch Signaling Pathway

Objective To investigate the effect of Qinghua Gushen Paidu Decoction on possible mechanism of renal protection and renal interstitial fibrosis in unilateral ureteral obstruction rats and Notch Signaling Pathway.We established a rat model of unilateral ureteral obstruction,and expect to find zhe theoretical support for anti-renal fibrosis and renal protection by Qinghua Gushen Paidu Decoction.Methods 72 male(Specefic pathogen Free,SPF)SD rats were randomly divided into 6 groups which sham-operation group(n=18),model group(n=18),Lotensin group(n=18)and Qinghua Gushen Paidu Decoction(n=18),and establish the model of UUO procedure.At 7,14,21 days,6 rats in each group were sacrificed.Blood samples were collected from herat,then measured serum urea nitrogen and serum creatinine.The pathological of the obstruction renal tissue were examined by HE and Masson staining.The protein expression levels of TGF-?1,?-SMA,Notch1 and Jagged1 were observed by immunohistochemical method.Quantitative real-time PCR was adopted to inspect the m RNA expression of Notch1 and Jagged1.2.SPSS23.0 was used.Data were analyzed using one-way ANOVA.All values were expressed as mean±SE,statistical significance was assigned at a p value of <0.05.Result1.The results of 24 hours urinary protein : There was no significant difference of 24 hours urinary protein in sham operation group,model group,Lotensin group and Qinghua Gushen Paidu Decoctiongroup(P > 0.05).2.The results of serum creatinine and urea nitrogen: Compared with the sham-operation group,serum creatinine and blood urea nitrogen significantly increased in the model group,Lotensin group and Qinghua Gushen Paidu Decoction group.(P<0.05).Compared with the model group,the levels of serum creatinine and urea nitrogen significantly decreased in the Lotensin group and Qinghua Gushen Paidu Decoction group on the 7th,14 th and 21 st days(P<0.05).There was no significant difference abou the levels of serum creatinine and urea nitrogen between Lotensin group and Qinghua Gushen Paidu Decoction group on the 7th,14 th and 21 st days(P> 0.05).3.The results of HE staining : The glomerular structure of the sham operation group was complete.There was no obvious infiltration of inflammatory cells in the interstitium.In the model group,there were diffuse vacuolar degeneration of some renal tubular epithelial cells,dilated lumen,atrophy and necrosis of renal tubular epithelial cells,fibroblast proliferation and interstitial fibrosis in renal interstitium.Compared with the UUO model group,the rats renal tubular injury degree of Lotensin group and Qinghua Gushen Paidu Decoction group has significantly reduced.The visible part of the structural integrity of the renal tubules,renal interstitial infiltration of inflammatory cells decreased.4.The results of Masson staining : Renal tissue of sham operation group visible blue collagen fiber trace,renal tissue morphology was normal,the basic rules of glomerular morphology,renal tubules arranged closely;renal tissue of the rats in the model group can be seen a large number of blue collagen fiber distribution,with the obstruction time prolonged,the blue collagen fibers increased significantly said,the degree of renal fibrosis is gradually increased.5.The results of immunohistochemistry : Renal tissue of sham operation group in the renal interstitium showed a low positive expression of brown,a brown positive expression in renal interstitial in model group,Lotensin group and Qinghua Gushen Paidu Decoction group in the renal interstitial showed moderate positive expression of brown.Compared with the sham operation group,TGF-?1,?-SMA,Notch1 and Jagged1 expression of model group,Lotensin group,Qinghua Gushen Decoction group significantly increased on the 7th,14 th and 21 st days(P < 0.05).Compared with the model group,TGF-?1,?-SMA,Notch1 and Jagged1 expression of Lotensin group,and Qinghua Gushen Decoction group decreased significantly on the 7th,14 th and 21 st days(P<0.05).There was no statistically significant difference about expression of TGF-?1,?-SMA,Notch1 and Jagged1 between Lotensin group and Qinghua Gushen Paidu Decoction group on the 7th,14 th and 21 st days(P > 0.05).6.The results of real time fluorescence quantitative PCR detection: Compared with sham operation group,Notch1 m RNA,Jagged1 m RNA expression of model group,Lotensin group and Qinghua Gushen Decoction group increased significantly(P<0.05).Compared with the model group,Notch1 m RNA,Jagged1 m RNA expression of Lotensin group and Qinghua Gushen Decoction group decreased(P < 0.05).Conclusion 1.Qinghua Gushen Paidu Decoction can delay to renal function deterioration of unilateral ureteral obstruction model in a certain extent,and reduce levels of serum creatinine and urea nitrogen,and protect kidney.2.According to reduing possibly of TGF-?1 and ?-SMA protein expression,Qinghua Gushen Paidu Decoction achieve its anti-renal fibrosis and renal protection.3.The overexpression of Notch signaling pathway leads to the progression of renal fibrosis in rats with unilateral ureteral obstruction model.Qinghua Gushen Paidu Decoction can inhibit renal interstitial fibrosis and renal interstitial fibrosis,which may be related to the inhibition of Notch signaling pathway.