| Objective To investigate the intervention effect of kaijie decoction on the signal pathway of Wnt/?-catenin in rats with adenine induced renal fibrosis and the mechanism of kaijie decoction to improve the inflammatory state of renal fibrosis.Methods Based on the success of the preliminary experiment,70 SPF SD male rats were randomly divided into the normal group(10 rats)and the model group(60 rats)according to their weight.After 21 days of continuous gavage,they were randomly divided into 5 groups:model group,high dose group,medium dose group,low dose group and postive control group.The treatment lasted for 4weeks.Then the rats were placed into the metabolic cage and collect urine for 24 hours then centrifuged to extract the supernatant.Anesthetized rats,5ml blood was taken from the apex of the heart and the serum was extracted after centrifugation.Take the kidney,observe the size,texture and color of the kidney,after cleaning,quickly put into 4% paraformaldehyde fixed.Serum Urea and Scr were tested by enzyme-motility assay;24hUP was detected by immunoturbidimetry;?1-MG was determined by ELISA.HE staining was used to observe the pathological changes of renal tissue;Masson staining observed collagen deposition;The expression of E-cadherin and ?-SMA in rat kidney were detected byimmunohistochemistry;Use the Western Blot and Real-time Quantitative PCR Detecting System to detect the expression of wnt1,wnt4,?-catenin,MMP-7,c-Myc and Col?rat kidney.Results 1.Effect of histopathological changes(1)HE staining results showed that in the blank control group did not observe the pathological changes in the renal tissues.In the model group,a large number of dark brown crystals were deposited in the renal tubules,which were damaged and infiltrated by a large number of inflammatory factors.After 4 weeks of drug treatment,compared with the model group the pathological changes of each treatment group were significantly improved.(2)MASSON staining result showed that compared with the normal group,the kidney collagen area of the model group increased significantly(P<0.01).After 4weeks of treatment,compared with the model group,the renal collagen area of each treatment group were significantly reduced(P<0.01).2.Determination of serum Scr and UreaCompared with the normal group,the levels of serum Scr and Urea in the model group were significantly higher(P<0.05),After drug intervention,the levels of Scr and Urea in serum of rats in all treatment group were significantly reduced(P<0.05).3.Result of the 24-hour urinary protein and ?1-MGCompared with the normal group,the contents of 24 UP and ?1-MG in the model group were significantly increased(P<0.05).After the drug intervention,the contents of 24 UP and ?1-MG in all treatment groups were significantly reduced(P<0.05).4.Immunohistochemical resultsCompared with the normal group,the expression of E-cadherin protein inkidney tissues of the model group was significantly decreased(P<0.01).After drug intervention,compared with the model group,the expression of E-cadherin protein in the kidney tissues of rats in the high-dose kaijie decoction group and the postive group increased significantly(P<0.05)Compared with the normal group,the expression of ?-SMA protein in kidney tissues of the model group was significantly increased(P<0.01).After drug intervention,compared with the model group,the expression of ?-SMA protein in the kidney tissues of the kaijie decoction high-dose group,medium-dose group and the postive group was significantly reduced(P<0.05)..5.Western bolt resultsCompared with the normal group,the expression levels of wnt1,wnt4,?-catenin,mmp-7,c-myc and Col?protein expression levels in the model group were significantly increased(P<0.01).After the drug intervention,the wnt1,?-catenin and Col?in the kaijie decoction high-dose group,the medium-dose group and the postive control group protein expression level decreased obviously(P<0.05 or P<0.01).The expression levels of wnt4,mmp-7 and c-myc in kaijie decoction high-dose group,the medium-dose group,the low-does group and the postive control group were significantly decreased(P<0.05 or P<0.01).6.qPCR test resultsCompared with the normal group,the mRNA levels of wnt1,wnt4,?-catenin,mmp-7,and c-myc in the model group were significantly increased(P<0.01).Compared with the model group,the mRNA levels of wnt1,wnt4,?-catenin,mmp-7,and c-myc in the kaijie decoction high-dose group,the medium-dose group,the low-dose group and the postive control group were significantly decreased(P<0.05 or P<0.01).Conclusion 1.Kaijie decoction can improve the status of nephritis in rats with renal fibrosis caused by adenine,and has a certain protective effect on the kidney.2.Kaijie decoction can effectively reduce the damage of renal fibrosis by intervening the signal pathway of Wnt/ ?-catenin in rats with adenine induced renal fibrosis.This effect has a certain dose-effect relationship,among which kaijie decoction has the best effect in the high-dose group and the medium-dose group. |
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