| Objective: Rat renal interstitial fibrosis model was prepared by unilateral ureteral obstruction(UUO)and valsartan was used as a positive control drug to study the effect of kudiezi injection on renal interstitial fibrosis and further observation the effect of kudiezi injection on superoxide dismutase(SOD)activity,malondialdehyde(MDA)content,as well as the expression of transforming growth factor-β1(TGF-β1),phosphorylated-Smad2/3(p-Smad2/3),bone morphogenetic protein-7(BMP-7),type Ι collagen protein and their genes,α-SMA protein in renal tissue of rats in each group.Methods: Sixty male SD rats were randomly divided into three groups,including the normal group and the sham operation group(ten rats in each group)and the modeling group(forty rats).The renal interstitial fibrosis model was established by ligating the left ureter of the rat.After the model was successful,the modeling group was randomly divided into model group,valsartan group,low dose group and high dose group of kudiezi injection(ten rats in each group).All rats were administered on the first day after operation.Valsartan group was intragastrically administered with 0.01g/(kg●d),low dose group and high dose group of kudiezi injection were intraperitoneally injected with 0.15g/(kg●d)and 0.30g/(kg●d),respectively.The remaining groups were intraperitoneally injected with the same volume of normal saline and the rat was weighed at intervals of 2 days to adjust the dosage.After seventh days and fourteenth days of UUO,blood and left renal tissue of rats were collected.The serum creatinine and blood urea nitrogen level were detected.HE staining,Masson staining and calculation of tubulointerstitial damage index,renal interstitial collagen deposition in the relative area score were performanced.SOD activity and MDA content in the kidney were measured by colorimetry.The expression of TGF-β1,p-Smad2/3,BMP-7,α-SMA protein in renal tissue of rats were detected with immunohistochemical staining and the content of TGF-β1,α-SMA,type Ι collagen(ColΙ)were detected by Western Blot.The expression of TGF-β1,Smad2,Smad3,BMP-7,ColΙ m RNA in renal tissue of rats were detected by reverse transcriptase polymerase chain reaction(RT-PCR).Results:1.Compared with the normal group and the sham operation group,the level of serum creatinine in model group rats was increased significantly;compared with the model group,valsartan and kudiezi after intervention could reduce the serum creatinine level(P<0.05);the rats blood urea nitrogen level after unilateral ureteral obstruction increased,but there was no difference among groups(P>0.05).2.HE staining and Masson staining showed that the morphology and structure of glomeruli and renal tubules in the normal group and the sham operation group were normal.On the seventh day after unilateral ureteral ligation,the renal tubular epithelial cells were partially exfoliated,vacuolar degeneration of cells,partial lumen dilation or atrophy,interstitial edema,infiltration of inflammatory cells,and focal interstitial fibrosis in the model group.On the fourteenth day after unilateral ureteral ligation,most of the structures of renal tubules and interstitium disappeared,and the remaining renal tubule lumen was diffuse or atrophic,a large number of inflammatory cell infiltration and diffuse interstitial fibrosis of model group.The index of renal tubulointerstitial damage and the relative area score of collagen fiber increased significantly.Compared with the model group,the renal tubular epithelial cell lesion,interstitial fibrosis in all drug treatment groups were alleviated,tubulointerstitial damage index and the relative area of collagen fiber decreased(P <0.05 or P<0.01).The antifibrosis effect was equal to the kudiezi high dose group and the valsartan group.3.Compared with the normal group,the sham operation group,the activity of SOD decreased and the content of MDA increased in the left renal tissue of the rat in the model group after unilateral ureteral ligation.Compared with the model group,the content of MDA decreased and the activity of SOD increased(P<0.01 or P<0.05)after the treatment of kudiezi and valsartan.There was a difference between the high and low dose groups of the kudiezi(P<0.05).4.Compared with the normal group and the sham operation group,the expression level of TGF-β1,p-Smad2/3 and α-SMA of the model group increased significantly,and the expression of BMP-7 significantly decreased in the left kidneys of rats(P<0.01).Compared with the model group,kudiezi and valsartan could reduce the expression of TGF-β1,p-Smad2/3 andα-SMA and increased the expression of BMP-7 in rats renal tissue(P<0.05 or P<0.01).5.Compared with the normal group and the sham operation group,the expression of TGF-β1,Smad2,Smad3,Col I m RNA significantly increased in left renal tissue of rats in the model group(P<0.01).Compared with the model group,the expression of TGF-β1,Smad2,Smad3 m RNA decreased and the expression of BMP-7 m RNA increased in the renal tissue after the drug treatment.Conclusion:1.Kudiezi injection can reduce the level of blood creatinine in rats and protect the renal function of UUO rats.2.Kudiezi injection can reduce the injury of tubulointerstitial and the deposition of collagen fibers in renal interstitial of UUO rats,indicating that it has the effect of resisting renal interstitial fibrosis to some extent.In terms of the effect of fibrosis,the high dose group of the kudiezi injection is equivalent to the positive control drug valsartan.3.Kudiezi injection can alleviate renal interstitial fibrosis in UUO rats,which may be related to the reduction of oxidative stress level in rat renal tissues and the regulation of the expression of key proteins in the TGF-β1/Smad/BMP-7 signaling pathway and reduction of proliferation and activation of myofibroblast in renal interstitium. |
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