The Effects And Mechanisms Of Anisodine Hydrobromide On Renal Interstitial Fibrosis In UUO Rats

Objective: The animal model of renal interstitial fibrosis(RIF)was established by the method of unilateral ureteral obstruction(UUO)and use this animal model to explore the effects and mechanism of Anisodine HydrobromideMethods: 40 SD male rats were randomly divided into four groups including:sham operated group,UUO model group(model group),UUO model treated with high doses Anisodine Hydrobromide(high dose group),UUO model treated with low doses Anisodine Hydrobromide(low dose group).Each group have 10 rats.Since the operation day,high dose group were given 5mg/kg·d Anisodine Hydrobromide by peritoneal injection once a day,low dose group group were given 2.5mg/kg·d Anisodine Hydrobromide by peritoneal injection once a day,the operated group and model group were given Isometric saline by peritoneal injection once a day.14 days later,the rats were sacrificed.Blood samples and renal tissue were collected.Serum creatinine(SCR)and urea nitrogen(BUN)were measured by automatic biochemical analyzer.Renal tissue was observed to assess renal pathological changes.The relative percentage of renal tubulointerstitial injury and renal interstitial fibrosis were assess by HE and MASSON stain.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in renal tissue were detected by WST-8 method and thiobarbituric acid method.Immunohistochemical staining and Western blot were used to detect the expression of transforming growth factor-beta(TGF-beta)and alpha smooth muscle actin(?-SMA)in renal tissue.Results:1 Body weight : Compared with the sham operated group,the weight of rats in other groups were reduced to varying degrees.Compared with the model group,the weight of high dose and low dose group increased in different degrees,which was the most obvious after 14 days(P < 0.05).There was no significant differences between high dose group and low dose group(P >0.05).2 Renal function: Compared with sham operated group,the levels of SCR and BUN in other groups were increased in different degrees.Compared with the model group,the levels of SCR and BUN in high and low dose groups were lower than those in the model group.There was no significant differences between high dose group and low dose group(P>0.05).3 Pathological changes in kidney: HE and Masson staining showed that there were no significant pathological changes in renal glomerulus,renal tubules and renal interstitium in sham operated group.In the other groups,the renal cortex became obviously thinning,and showed varying degrees of lumen expansion,tubular epithelial cells atrophy,interstitial inflammatory cell infiltration and hyperplasia of fibrous tissue.The tubulointerstitial injury index and the relative area of the renal collagen score were significantly increased in other groups(P < 0.05).There was no significant differences between high dose group and low dose group(P>0.05).4 The levels of SOD and MDA in renal tissue: Compared with sham operated group,the level of SOD in renal tissue of model group was significantly decreased(P<0.05)while the level of MDA was significantly increased(P <0.05).Compared with the model group,the levels of SOD in the two Anisodine Hydrobromide treat groups were increased,and the levels of MDA were decreased,among which,the changes of the high dose group was more significant(P < 0.05).There was no significant differences between high dose group and low dose group(P>0.05).5 Immunohistochemical results of TGF-?1: In sham operated group,the expression of TGF-?1 was only weakly expressed in the renal tubular epithelium.The expression of TGF-?1 was increased in other groups located in renal tubule and interstitium(P < 0.05).After treated with Anisodine Hydrobromide,the expression of TGF-?1 was significantly decreased(P <0.05).There was no significant differences between high dose group and low dose group(P>0.05).6 Immunohistochemical results of ?-SMA: In sham operated group,there was almost no expression of ?-SMA in renal interstitial.In other groups7 Western blot results of ?-SMA?TGF-?1 in renal tissue: A little expression of?-SMA ? TGF-?1 were observed in operated group while the expression of?-SMA?TGF-?1 were increased in model groups(P<0.05).After treated with Anisodine Hydrobromide,the expression of ?-SMA ? TGF-?1 were significantly decreased(P < 0.05).There was no significant differences between high dose group and low dose group(P>0.05).Conclusion:1.Anisodine Hydrobromide can improve the renal interstitial fibrosis in UUO rat model2.Oxidative stress is involved in the pathogenesis of renal interstitial fibrosis,the effect of Anisodine Hydrobromide is associated with oppose oxidative stress.3.Anisodine Hydrobromide may inhibit the expression of TGF-? and reduce the activation of fibroblasts,so as to improve the renal interstitial fibrosis.