| OBJECTIVE:To observe the effect of Taohe Chengqi Decoction on miR-29 expression mediated by TGF-β1/Smad3 signaling pathway in renal interstitial fibrosis in chronic renal failure(CRF)rats,and to analyze the feasibility of delaying CRF by this party.Provides molecular biology basis for the treatment of CRF.METHODS:One hundred adult male Wistar rats,7 weeks old,were fed adaptively for 1 week and randomly divided into four groups(n=25).They were named blank group,sham group,model group,and treatment group.Rats in both the model group and the treatment group were modeled with 5/6 nephrectomy,and the kidneys of the left kidney 2/3 were excised under chloral hydrate anesthesia.One week later,a second operation was performed to remove the right renal tissue from the rat;in the sham-operated group,only the two kidneys were removed;the blank group was not treated.Penicillin was injected intraperitoneally three days after surgery to prevent infection.Each group was fed for 4 weeks after the completion of the modeling,and each group of rats was given intragastric administration(10 ml/Kg-d)every morning for 4 weeks.Rats in the blank group,sham operation group,and model group were given intragastric administration with normal saline.The treatment group was lavaged with Taohe Chengqi Decoction and was administered by continuous intragastric administration for 56 days.The abdominal aorta was used to take blood for detection of RBC,Hb,Scr,and BUN in rats;the left renal tissue was taken and part of the kidney tissue was placed in cryovials.Western Blot was used to detect the protein levels of TGF-β1 and Smad3 in the left kidney;Real-Time PCR was used to detect the expression of TGF-β1 Smad3 mRNA and miR-29a,miR-29b,and miR-29c;the other part was stored in kidney tissue.In paraformaldehyde,after treatment,the pathological conditions of each group of kidney tissues were observed with a light microscope and pictures were taken to compare the changes of each group.RESULTS:Compared with the blank group and the sham operation group,the contents of RBC and Hb in the model group,the treatment group,and the sham operation group all decreased to different degrees(P<0.05).The serum SCr and BUN increased to different degrees(P<0.05).The results of the treatment group compared with the model group were statistically significant.Western Blot results showed that the expression of TGF-(31 and Smad3 protein in the left kidney was significantly decreased in the treatment group compared with the model group(P<0.05);the expression of TGF-β1 and Smad3 mRNA in the treatment group compared with the model group was shown by Real-Time PCR Down-regulation(P<0.05),miR-29a,miR-29b,and miR-29c expression levels were up-regulated(P<0.05);HE staining of renal interstitial fibrosis under light microscope showed left kidney structure in the blank group and sham-operation group.Normal;Model group left kidney structure disorder,severe interstitial fibrosis,renal tubular atrophy,visible inflammatory cell infiltration;treatment group compared with the model group,renal interstitial inflammatory infiltration reduced interstitial fibrosis significantly improved.CONCLUSION:1.Taohe Chengqi Decoction can effectively improve renal anemia in CRF rats;significantly reduce serum creatinine and urea nitrogen levels in CRF rats and effectively improve renal function in rats with chronic renal failure;can protect CRF Rat kidneys delay the progression of chronic fibrosis;TGF-β1/Smad3 signals are activated in 2.5/6 nephrectomized rat renal interstitial fibrosis model;3.Taohe Chengqi Decoction can inhibit 5/6 nephrectomized rats The TGF-β1/Smad3 signal in the renal interstitial fibrosis model up-regulated the expression of miR-29. |
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