The Role Of TianQi Injection Against Kidney Fibrosis And The Mechanism Of TGF-β1/Smads Signaling Transduction

Objective:1. To observe the effect of TianQi injection on the structure, fuction and Extracellular matrix of kidney, confirming the anti-fibrosis role of TianQi injection.2. To investigate the anti-fibrosis mechanism of TianQi injection on the TGF-β1/Smads signaling conduction.Methods:1. The adriamycin-induced kidney fibrosis models were established by ways of excising right kidney and injecting Adriamycin into vena caudalis.2. The 48 healthy male Sprague-Dawley (SD) rats were randomly divided into four groups:normal group, sham group, model group, TianQi injection treatment group. The rats in the normal group feed under normal condition without experienceing the operation were dealed with normal sodium(NS) (10 ml/kg·d) by intraperitoneal injection; The rats in the sham group after remove right kidney were dealed with normal sodium(NS) (10 ml/kg·d) by intraperitoneal injection without receiving adriamycin;The rats in the model group received NS (10 ml/kg·d) by intraperitoneal injection after operation; The rats in the TianQi injection treatment group received TianQi injection (10 ml/kg·d) by intraperitoneal injection after operation; All the rats were sacrificed after receiving treatments for 4 weeks, respectively, and the urine, blood and kidneys were collected.24h quantity of urinary protein and kidney function were detected; Kidney histological examinations was performed by HE and Masson staining, and the degree of tubular-interstitial damage was valuated in terms of semi-quantitative score. The mRNA and/or protein expression of TGF-β1, TGF-βR-Ⅰ, TGF-βR-Ⅱ, p-Smad2 anc p-Smad7 were detected by means of reverse transcription-Polymerase chair reaction (RT-PCR), inununohistochemistry and Western blot.Results:1. After excising right kidney and injecting Adriamycin into vena caudalis 24h quantity of urinary protein in rats was increased (P<0.01), and the BUN and Scr level in serum sample was enhanced (P<0.01), and the kideny pathology such as glomcrulus sclerosis, tubular atrophy, interstitial fibrosis and inflammation cells infiltration were found in rats, it suggests a successful kidney fibrosis model.2. When compared with sham group, the urinary protein in model group rats was increased (P<0.01), and the BUN and Scr level in serum sample was enhanced (P<0.01). The protein expression of CL-IV and a-SMA in rats model witl Adriamycin-induced kidney fibrosis were increased when compared with sham group (P<0.01), which suggests an EMT course occuring in kidney fibrosis rats TianQi injection treatment could reduce the quantity of urinary protein (P<0.01) lower the BUN and Scr level(P<0.01), ameliorate the pathology in kidney such a glomcrulus sclerosis, tubular atrophy, interstitial fibrosis and inflammation cells infiltration, reduce the scores of glomcrulus sclerosis area and interstitia fibrosis(P<0.01),down-regulate the protein expression of collagen type IV(P<0.01 when compared with model group, which indicates that TianQi injection plays anti-kidney fibrosis role, and the mechanism is involved in influencing the course o EMT.3. The mRNA and/or protein expression of TGF-β1, TGF-βtypeⅠreceptor, TGF-βtypeⅡreceptor, p-Smad2 and p-Smad7 in Adriamycin-induced kidney fibrosis models were up-regulated when compared with sham group (P<0.01),which suggests that the TGF-β1/Smads signaling is activated. When compared with model group, TianQi injection treatment could significantly down-regulate the mRNA and/or protein expression of TGF-β1, TGF-βtypeⅠreceptor, TGF-P typeⅡreceptor, p-Smad2 (P<0.01), and up-regulate protein expression of p-Smad7 (P<0.01), which indicates that anti-kidney fibarosis role of TianQi injection is associated with repressing the activation of TGF-β1/Smads signaling.Conclusion:1. The activation of TGF-β1/Smads signaling in the adriamycin-induced kidney fibrosis models is involved in the course of EMT in the tubular.2. TianQi injection plays a anti-fibrosis role that is perhaps associated with repressing TGF-β1/Smads signaling mediated tubular EMT.