| Objective:To establish a model of renal interstitial fibrosis in rats with unilateral ureteral obstruction(UUO)and observe the effect of Ginkgo biloba extract(GBE)on tr ansforming growth factor β1 and bone morphogenetic protein in renal tissue of renal int erstitial fibrosis.The influence of expression was to explore the intervention effect and p ossible mechanism of Ginkgo biloba extract in the process of renal fibrosis,to clarify th e mechanism of Ginkgo biloba extract on renal protection.Methods:72 male SPF Wistar rats(body weight about 200±10g,6 weeks old)w ere randomly divided into 3 groups,24 in each group.Group A was normal control gro up(control group,Sham group),M group was UUO model group(model Group,UUO group),and group R was Ginkgo biloba extract treatment group(treatment group,GBE group).The obstructed renal tissues were randomly selected from 3rd,7th,and 14 th da y after unilateral ureteral obstruction respectively,in the N3,N7,and N14 groups,with 8 in each group.Rats in group M and group R were anesthetized by intraperitoneal injection of 10%chloral hydrate.Under conventional disinfection,there were incisions made of 1.5 cm o n side of the left back spine and 1 cm in the longitudinal incision under the ribs.The s kin,subcutaneous tissue,muscle,peritoneum and visceral peritoneum were incised in lay er by layer.Looking for the left ureter along the lower left kidney,4 # line double liga ture ligation of the upper 1/3 of the left ureter,and then cut from it.Rats in group A only detached the ureter and did not cut the ureter.The rest of the procedure was the s ame as before.At the end of the 3rd,7th and 14 th day,the rats were disposed in batc hes.The renal tissues were taken out.The pathological changes were observed by HE a nd Masson staining.The expressions of BMP-7 and TGF-β1 in the kidney tissues of r ats were observed by immunohistochemical staining.Semi-quantitative analysis was perfo rmed using image analysis system software Image-Pro Plus6.0.The treatment group received 100 mg/kg/d ginkgo biloba extract,control group and the model group were administered with 0.9% normal saline once a day.Rats were ga vaged a day before the molding surgery.During the experiment,rats in each group wer e given free access to water,standard diet,and grown in a constant temperature and hu midity environment.Results: 1.HE staining: Results of HE staining: On the 3rd day after operation in UUO group,renal tubular injury began to appear in the renal tissue of rats.As the obst ruction time prolonged,the renal tubules expanded progressively,inflammatory cell infiltr ation gradually increased,and some tubules atrophied.The damage increased and the tub ulointerstitial injury score(TIS)gradually increased,the difference was statistically signifi cant(P<0.05).There was no obvious pathological change in the renal tissue of Sham gr oup.Compared with the time point corresponding to Sham group,TIS increased in UU O group,the difference was statistically significant(P<0.05).Compared with the time po int of UUO group,GBE group renal tubule.The injury was alleviated and the TIS was decreased.The difference was statistically significant(P<0.05).2.Masson staining results: On the 3rd day after operation,the interstitial fibers beg an to increase in the UUO group.As the obstruction time prolonged,renal interstitial co llagen fiber deposition area gradually increases,and fibrous connective tissue with diffus e hyperplasia appeared.The area ratio of fibrosis(RIF index)gradually increased,and th e difference was statistically significant(P<0.05).The Sham group interstitial collagen fi bers had a small green staining area and no renal interstitial fibrosis.Compared with the time points corresponding to the Sham group,the renal interstitial fibrosis(RIF)index of the UUO group increased,and the difference was statistically significant(P<0.05).Co mpared with the time points corresponding to the UUO group,the degree of interstitial fibrosis in the GBE group was improved to some extent,and the RIF index was decrea sed,the difference was statistically significant(P<0.05).3.Immunohistochemical results:(1)Immunohistochemistry showed that the expressio n of TGF-β1 protein was mainly located in renal tubular epithelial cells.With the prolo ngation of obstruction time,the expression of TGF-β1 protein in renal tissue of UUO gr oup increased gradually.The difference of time points was statistically significant(P<0.05).Compared with the time point corresponding to Sham group,the expression of TGF-β1 protein increased gradually,the difference was statistically significant(P<0.05);the ti me corresponding to UUO group Compared with the point,the expression of TGF-β1 pr otein in GBE group decreased,the difference was statistically significant(P<0.05).(2)I mmunohistochemistry showed that BMP-7 protein was mainly expressed in renal tubular epithelial cells and Renal interstitial.The expression of BMP-7 protein in UUO group decreased gradually with the prolongation of obstruction time.The difference of time poi nts was statistically significant(P<0.05).Compared with the time point corresponding to Sham group,the expression of BMP-7 protein in UUO group decreased,the difference was statistically significant(P<0.05);compared with the time point corresponding to U UO group,GBE group BMP-7 protein expression increased,the difference was statistic ally significant(P<0.05).Conclusions:1.In UUO rat renal interstitial fibrosis,TGF-β1 protein expression in creased,suggesting that TGF-β1 is involved in the progression of renal interstitial fibrosi s.2.After applying Ginkgo biloba extract,the expression of TGF-β1 protein decreased,the expression of BMP-7 protein increased,and the degree of renal fibrosis decreased.It indicated that Ginkgo biloba extract has renal protective effect. |
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