Yiqihuoxue Jiangzhuo Side Of Renal Interstitial Fibrosis Of Tgf-beta <sub> 1 </ Sub> The / Smads/bmp-7 Signal Transduction Pathways Affect Experimental Research

Glomerular sclerosis and tubular interstitial fibrosis is the common performance and main pathological basis of which all sorts of chronic kidney diseases progress to end-stage kidney failure. In recent years, tubular interstitial fibrosis have increasingly played a significant role in the progress of chronic kidney diseases. Therefore, to discuss the prevention measures of renal interstitial fibrosis are of great significance in clinical. Chinese medicine have achieved good effects in preventing organs fibrosis aspects and also have shown broad application prospects. This experiment adopts Yiqi Huoxue Jiangzhuo Formula(YQHXJZF) which is composed by astragalus mongholicus, angelica, rhizomaligusticiwallichii, white paeonia root, radix bupleuri, baikal skullcap root, rhubarb, tuckahoe and has the efficacy of tonifying qi and activating blood, clearing away dampness and dropping turbidity. According to Chinese medicine's pharmacology function of multicomponent, multi-link,multi-level and multi-target for preliminary exploration of the treatment function of YQHXJZF with renal fibrosis, we established unilateral ureter obstruction to cause renal fibrosis in.the rat model and used immunohistochemistry, real-time quantitative PCR to conduct a research from organization, protein, gene level. It aimed to preliminarily probe into YQHXJZF's possible mechanism of action of preventing and curing RIF.Objective:In.order to clarify the possible mechanism of YQHXJZF in preventing RIF from levels'of tissue, protein and gene, we established UUO rat model and then observed the efficiency of YQHXJZF in influence TGF-β1/Smads/BMP-7 signal transduction pathways and related factors'expression as well as studied if YQHXJZF can improve renal function of UUO rat model in a certain degree or reduce kidney pathological damage.Methods:We adopted unilateral ureter obstruction to cause renal fibrosis in the rat model and the rats were randomly divided into seven groups:normal control group, sham operation group, model group, Fosinopril group, YQHXJZF high-dose group, YQHXJZF medium-dose group and YQHXJZF low-dose group.Each group rats were executed on the 7th,14th,21th day and the following indexes were detected.1. Pharmacodynamic:Observe the general condition and weekly weight of each group rats.Calculate the kidney weight/body weight ratio every week.The 24-hour urinary protein were detected by coomassie briliant blue method.After executing the rats every week, we separated serum of each rat to measure the serum biochemical indexes determination, such as blood urea nitrogen (Bun), serum creatinine (Scr), albumin(Alb).The HE and Mallory dyeing were used to observe the pathological morphology changes of the rats kidney.2. Immunohistochemistry:The immunohistochemical method was used to detect the expressions of TGF-β1, Smad6, BMP-7, CTGF, FN, ColⅠand ColⅢprotein.3. Real-time quantitative PCR:The real-time quantitative PCR was used to detect the expressions of TGF-β1mRNA, BMP-7mRNA, CTGFmRNA.Results:1. Pharmacodynamic:Yiqi Huoxue Jiangzhuo Formula(YQHXJZF) can improve the general condition of UUO rats to a certain extent but the effect on body weight is not very clear. Compared with the normal control group, the model group rats' kidney weight/body weight ratio increased significantly in different time points(P<0.01);the urine protein of model group also significantly increased (P<0.O1);After preparing mode, the level of Scr in model group also significantly rose in the second and third week(P<0.01) while the level of Alb declined (P<0.01), but there was no significant statistics difference in Bun. Compared with the model group, each treatment group rats'kidney weight/body weight ratio declined but there was no significant statistics difference in the first and second week;In the third week, Fosinopril group, YQHXJZF high-dose group, medium-dose group rats had significant statistics differences compared with the model group(P<0.05);The 24-hour urinary protein of each treatment group declined but there was no significant statistics difference in the first week; The 24-hour urinary protein of YQHXJZF high-dose group, low-dose group reduced(P<0.05) while the medium-dose group remarkably reduced(P<0.01) in the second week; In the third week, the 24-hour urinary protein of each treatment group significantly declined(P<0.01); The level of Scr, Bun and Alb in YQHXJZF medium-dose group, high-dose group and Fosinopril group had improved obviously in the third week(P<0.01, P<0.05).The glomerular sclerosis index and renal interstitial damage examination score in YQHXJZF medium-dose group, high-dose group also significantly reduced in different time points(P<0.01).2. The protein expressions of TGF-β1, Smad6, BMP-7, CTGF, FN, ColⅠand ColⅢin each group:The protein expressions of TGF-β1, CTGF, FN, ColⅠand ColⅢin model group were notably higher than those in normal control group in different time points (P<0.01), while YQHXJZF high-dose group, YQHXJZF medium-dose group, YQHXJZF low-dose group and Fosinopril group's expression significantly reduced(P<0.01); The protein expressions of Smad6, BMP-7 in model group were lower than those in the normal control group.3. The expressions of TGF-β1mRNA, BMP-7mRNA, CTGFmRNA in each group:Compared with the normal control group, the expressions of TGF-β1mRNA, CTGFmRNA in model group significantly up-regulated in different time points (P<0.01), while YQHXJZF medium-dose group, YQHXJZF high-dose group and Fosinopril group significantly down-regulated the mRNA expressions of TGF-β1 and CTGF in different time points(P<0.01, P<0.05). Just the opposite happened in the expression of BMP-7mRNA.Conclusions:1. Yiqi Huoxue Jiangzhuo Formula(YQHXJZF) can improve the general condition of UUO rats to a certain extent but the effect on body weight is not very clear. It also can reduce the kidney weight/body weight ratio, and decline the 24-hour urine.protein excretion, and improve the level of serum biochemical, indexes, such as Scr, BUN, ALB.2. YQHXJZF can alleviate severe pathological changes of kidney in rats with UUO, such as relieve the renal tubular epithelial cells falling off and vacuolar degeneration,alleviate renal interstitial inflammatory cells infiltrating, relieve lumens of renal tubules expanding and the less integrity of renal tubular basement membrane's structure.3. YQHXJZF can down-regulate the protein expressions of TGF-Pi, CTGF, FN, ColⅠand ColⅢ, and up-regulate the protein expressions of BMP-7, Smad6.4. YQHXJZF can down-regulate the mRNA expressions of TGF-β1 and CTGF as well as up-regulate the mRNA expression of BMP-7.In a word, YQHXJZF has a certain protective effect in the kidney of UUO rats. Its mechanism is likely regulated by TGF-β1/Smads/BMP-7 signaling transduction pathway:YQHXJZF might inhibit the over-expression of CTGF through down-regulation the expression of TGF-β1 as well as up-regulation the expressions of Smad6, BMP-7, and prevent the synthesis of collagen and extracellular matrix, thus playing the role of antagonisting renal interstitial fibrosis.