| Objectives Establishing the model of rats by the operation of two Kidney-one Clip, the pathological changes of the rat's heart and kidney tissues were detected, and then explore the effect of Rhein lysinate(RHL) on the fibrosis of kidney and heart tissues of the 2k1 c deduced hypertensive rats, and elucidate the possible molecular mechanism of RHL in inhibition of fibrosis of heart and kidney tissues in rats, and might provide the reference of RHL in reducing the renovascular hypertension and inhibiting the process of heart and kidney tissues fibrosis.Methods 1 Modeling: 35 male Sprague-Dawley(SD) rats are needed in this experiment,28 rats were established into two-kidney one-clip(2K1C) renovascular hypertensive rat models,and the other 7 rats were divided into control group(control group is the same as the shame group). Rats' systolic blood pressures(SBPs) of each group were measured, and the models were established successfully if SBP rised over20 mm Hg(1mm Hg=0.133 k Pa) and was higher ? 140 mm Hg.(26 rats were established successfully totally,the success rate is 93.33%.)2 Grouping: The number of rats of control group was 6,and 24 rats randomly chosen from 26 rats which were established successfully were divided randomly into control group, model group, RHL treatment group, rhein group and lysine group, 6 rats in each group. 3 Rats were put to death after the experiment groups had been established after 8 weeks. 4 Detecting:(1) SBPs of the rats in each group were measured before modeling, 4 weeks and 8 weeks after modeling.(2) The left ventricular weight(LVW), the body weight and the ratio of left ventricle weight to body weight(LVW/BW) were measured at the end point of the study.(3)Observing the pathological changes of kidney and heart tissue by HE staining.(4)Observing the fibrillation of kidney and heart tissue by Masson staining.(5) Observing the expression and localization of TGF-?1 and CTGF of kidney and heart tissue by immunohistochemical staining(IHC).(6) Observing the expression and localization of TGF-?1 and CTGF of kidney and heart tissue by immunofluorescence histochemistry staining(IF).Results 1 SBP variation:(1) compared to the SBPs of pre-operation, SBPs of model group and other 4 experiment groups rised over 140 mm Hg and the increment were over40 mm Hg 4 weeks after operation, the differences were statistically significant(P<0.05),which means that the models of two-kidney one-clip(2K1C) renovascular hypertensive rat were established successfully.(2)Comparasion of SBPs of each goup 8 weeks after operation: SBPs of the model group were obviously higher than that of the control group and the differences were statistically significant(P<0.05); compared to the model group,SBPs of the lysine group were not higher obviously, the differences were not statistically significant(P > 0.05); compared to the lysine group, SBPs of the RHL groups were decreased evidently, the differences were statistically significant(P<0.05); compared to the rhein group, SBPs of the RHL group were decreased evidently, the differences were statistically significant(P<0.05). 2 Comparasion of LVW/BW of each goup: compared to the control group, LVW/BW of the model group were higher obviously, the differences were statistically significant(P<0.05); compared to the model group, LVW/BW of the lysine group were not higher obviously, the differences were not statistically significant(P> 0.05); compared to the lysine group, LVW/BW of the RHL groups were decreased obviously, the differences were statistically significant(P<0.05); compared to the rhein group, LVW/BW of the RHL group were decreased obviously, the differences werestatistically significant(P<0.05). 3 HE staining: Kidney and heart tissues are nearly normal in the control group rats. Commpared to the control group, fibrous tissues were deposited in the kidney and heart tissues of the model group rats; Commpared to the model group,the fibrous tissues deposited in the kidney and heart tissues of the lysine group rats were not different obviously; Commpared to the lysine group, fibrous tissues deposited in the kidney and heart tissues of the RHL group rats were decreased obviously; Commpared to the rhein group, fibrous tissues deposited in the kidney and heart tissues of the RHL group rats was decreased obviously. 4 Masson staining:fibrous tissues deposited in the kidney and heart tissues are nearly normal in the control group rats. Commpared to the control group, fibrous tissues were deposited in the kidney and heart tissues of the model group rats(P<0.05); Commpared to the model group, fibrous tissues deposited in the kidney and heart tissues of the lysine group rats were not different obviously(P>0.05); Commpared to the lysine group, fibrous tissues deposited in the kidney and heart tissues of the RHL group rats were decreased obviously(P<0.05); Commpared to the rhein group, fibrous tissues deposited in the kidney and heart tissues of the RHL group rats was decreased obviously(P<0.05). 5 IHC staining: TGF-?1 and CTGF expressed in the kidney and heart tissues are nearly normal in the control group rats. Commpared to the control group, TGF-?1 and CTGF were expressed significantly in the kidney and heart tissues of the model group rats(P<0.05); Commpared to the model group, the difference of TGF-?1 and CTGF expression in the kidney and heart tissues of the lysine group rats were not obvious(P>0.05); Commpared to the lysine group, TGF-?1 and CTGF expressed in the kidney and heart tissues of the RHL group rats were decreased markedly(P<0.05); Commpared to the rhein group, TGF-?1 and CTGF expression in the kidney and heart tissues of the RHL group rats was decreased obviously(P<0.05).5 IF staining: TGF-?1 and CTGF expressed in the kidney and heart tissues are nearly normal in the control group rats. Commpared to the control group, TGF-?1 and CTGF were expressed significantly in the kidney and heart tissues of the model group rats(P<0.05); Commpared to the model group, the difference of TGF-?1 and CTGF expression in the kidney and heart tissues of the lysine group rats were not obvious(P>0.05); Commpared to the lysine group, TGF-?1 and CTGF expressed in the kidney and heart tissues of the RHL group rats were decreased markedly(P<0.05);Commpared to the rhein group, TGF-?1 and CTGF expression in the kidney and heart tissues of the RHL group rats was decreased obviously(P<0.05).Conclusions 1 The model of 2K1 C renovascular hypertensive rats can be established successfully. Fibrous tissues can be deposited in the kidney and heart tissues 8 weeks after modeling. 2 RHL played a role in decreasing the SBP of 2K1 C renovascular hypertensive rats significantly. 3 RHL might inhibit the fibrosis of rats of kidney and heart tissues.4. Fibrous tissues were deposited obviously in the kidney and heart tissues of 2k1 c rats, RHL can down regulate the expression of TGF-?1 and CTGF in rats of kidney and heart tissues. RHL may inhibit heart and kidney fibrosis by reducing TGF-?1 and CTGF expression. |
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