| Objective:By studying the effect of Taohe Chengqi Decoction on anemia,renal function,pathological morphology and expression of APC 、 β-catenin and TCF4 in Wnt/β-catenin signaling pathway in chronic renal failure(CRF)rats induced by 5/6 nephrectomy clinical medication to explore the mechanism of Taohe Chengqi Decoction in improving renal fibrosis in CRF rats,so as to provide objective experimental basis for the treatment of CRF and guide clinical medication.Methods:(1)125 clean 7-week-old male Wistar rats were fed with normal diet and ultrapure water for one week to make them adapt to the experimental environment.Firstly,50 rats were randomly selected and then divided them into blank group and sham operation group,with 25 rats in each group.The rats in the blank group were not intervened by operation and the rats in the sham operation group were only stripped of the two renal tunica;the remaining 75 rats adopted 5/6 nephrectomy to copy CRF model according to literature methods,that is,the left2/3 renal tissues were excised,then the right one was excised a week later;penicillin was injected 3 days after operation to prevent infection.After confirming that the 5/6nephrectomy model was successfully established,the rats were randomly divided into model group,Niaoduqing group and Taohe Chengqi Decoction group,with 25 rats in each group.(2)Dosage treatment:starting from the fourth week after the rats successful modeled,medication was given regularly at 8:00 a.m.every day: the rats in Niaoduqing group and Taohe Chengqi Decoction group were given Niaoduqing granular solution and Taohe Chengqi Decoction respectively,while rats in other three groups were given normal saline gavage.The dosage of rats was calculated as 6 times the dosage per kilogram of body weight of clinical adults,which was adjusted according to the weight change of rats.The general condition of rats was observed and recorded before daily gavage for 56 days.(3)Specimen detection: on the 57 th day,the rats were sacrificed after blood and residual renal tissue samples were taken.The blood specimens were used to detect RBC,HB,Scr and BUN;the kidney tissue specimens were used to detect the m RNA and protein expression of APC,β-catenin and TCF4 by the method of real-time FQ-PCR and Western Blot respectively;3 rats were respectively and randomly selected from each group,without taking blood,several pieces of 1 mm × 1 mm × 1 mm tissue were taken from the renal cortex,and then put into the precooled stationary liquid for the observation of renal pathological morphology by transmission electron microscope.Results:(1)The blood routine,renal function,m RNA and protein expression level and pathological morphology of the blank group and the sham operation group rats were normal and there was no significant difference between these two groups(P>0.05).(2)The levels of RBC and Hb in the Taohe Chengqi Decoction group rats were significantly higher than those in the model group rats(P<0.05).(3)The levels of Scr and BUN in the Taohe Chengqi Decoction group rats were significantly lower than the model group rats(P<0.05),and were also lower than the Niaoduqing group rats(P<0.05).(4)Compared with the model group and Niaoduqing group rats,the m RNA and protein expression levels of APC in Taohe Chengqi Decoction group rats were significantly increased(P<0.05),while the β-catenin and TCF4 expression levels were significantly decreased(P<0.05).(5)Transmission electron microscope showed that the ultrastructure of glomeruli and renal tubules in blank group and sham operation group rats were normal.In the model group rats,the ultrastructure of glomeruli and renal tubules could not be clearly identified under electron microscope;and there were a lot of inflammatory cells infiltration in capillaries,mesangial cells proliferation,foot process fusion,podocyte degeneration;renal tubular necrosis,epithelial cell edema,nuclear membrane damage in many places;mitochondria fuzzy,vacuolar changes;brush edge loss.The pathological changes of Niaoduqing group and Taohe Chengqi Decoction group rats were significantly milder than that of model group rats,and the integrity of glomeruli and renal tubular tissue in Taohe Chengqi Decoction group rats were better than that in Niaoduqing group rats.Conclusion:(1)Taohe Chengqi Decoction can significantly improve the renal anemia condition,renal function and renal pathological damage in the rat model of chronic renal failure.(2)Taohe Chengqi Decoction can reduce the degree of renal fibrosis in rats with chronic renal failure and delay the process of CRF.Its mechanism may be related to inhibiting the Wnt/β-catenin signal pathway,up-regulating the expression of APC and down-regulating that of β-catenin,TCF4. |
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