Based On The TGFf-β1-TIMPs-Col-Ⅳ Pathway To Explore Qian Yang Yu Yin Granules On SHR Intervention Effect Of Kidney Injury

Research Background:Long-term severe hypertension can cause renal fibrosis, induce renal injury. In our country,new imported2011dialysis patients, about9.9%of the primary disease is hypertension.The hypertensive renal injury is a serious threat to people’s health and quality of life.Qian Yang Yu Yin granules is that the professor Fang Zhuyuan inherit of Tang Shuhua’s academic experience, combined with his clinical practice, on the basis of clearing liver and improving fire, nourishing kidney and activating the blood circulation method.And it has been declared as hospital preparations in Jiangsu Provincial Hospital of Traditional Chinese Medicine (approval number:Su medicine system of word Z20100007).In the early clinical studies have shown that Qian Yang Yu Yin granules combined with Benazepril has better antihypertensive effect in patients with hypertensive renal injury, and decrease the urine protein, reduce AngⅡ levels, improve the function of kidney, significantly improve patients’ quality of life,but the specific mechanism of action has not been fully elucidated.The TGF-betal-TIMPs-Col-Ⅳ pathways activated in hypertensive renal injury, induced kidney damage, plays an important role in the process of hypertensive renal injury.TGF-betal is a multifunctional polypeptide growth factors, through a complex cell surface receptors signal pathways regulating cell proliferation differentiation and apoptosis.TGF-betal can stimulate the GMC synthesis of ECM components such as Col-Ⅳ,proteoglycan and so on,increase the ECM, promote glomerular sclerosis. And TGF-beta1has nudged to effect the biological characteristics of itself, the release of TGF-betal in injury can induce cells to produce more TGF-betal, thus its biology can be amplified.And TIMP-1MMP-9local maintain relative balance in the kidney, regulating kidney collagen proportion, keep the normal kidney structure and function.The experiment through the SHR renal damage induced by high salt diet model, by ELISA, Western blot method Real Time-PCR and so on, research that Qian Yang Yu Yin granules combined with Benazepril intervene effect of hypertensive renal impairment,and the TGF-betal-TIMPs-Col-IV pathway,provide experimental basis for the prevention and treatment of hypertension renal damage of traditional Chinese medicine, and provide ideas for future research. Research contents: The first part of theoretical research1. Research on the TCM theory of hypertensive renal impairment2.The modern medicine research of hypertensive renal impairment The second part of the experimental research12weeks of male WKY8as the normal control group;12weeks of male spontaneously hypertensive rats (SHR)32only according to completely random method is divided into four groups (each group of8), namely:hypertension model (SHR) Qian Yang yu Yin granules group,Benazepril group, Qian Yang Yu Yin granules+Benazepril group,all free water, for eight weeks, and examine the following indicators.1.Evaluation of Qian Yang Yu Yin granules on SHR early renal damage model of pharmacodynamics. Experiment of8weekend, noninvasive method of tail set measurement between groups of rats awake blood pressure changes before and after drug delivery;24h urine metabolism cage mouse access, using automatic biochemical analyzer test in rats urinary microalbuminuria and urinary NAG; HE staining evaluation of kidney disease change; Masson staining to evaluate renal fibrosis.2.Using Western Blot method to detect Qian Yang Yu Yin granules effect of TGF-betal, MMP-9,TIMP-1,Col IV protein expression in SHR kidney early renal damage model.3.1mmunofluorescence labeling kidney tissue TGF-betal and using Real time PCR detection of Qian Yang Yu Yin granules of the influence of TGF-betal mRNA gene expression in SHR early renal damage model. Result:1.Medication before, compared with other groups of SHR, WKY group were obvious statistical significance (P<0.01),SHR groups have no statistical significance (P>0.05), building success;8weeks after the treatment, and than WKY group, SHR model group significantly increased blood pressure, there is statistical significance (P<0.01); With SHR group than in Qian Yang Yu Yin granules groups,Benazepril group and Qian Yang Yu Yin granules+Benazepril group had a significantly lower blood pressure, there was statistically significant (P<0.01or P<0.05), which the effect of lowering blood pressure is especially combination group significantly (P＜0.01).2.Than WKY group, SHR model group obviously increased urinary microalbuminuria and urinary NAG, there is statistical significance (P<0.01); With SHR group than in Qian Yang Yu Yin granules groups,Benazepril group and Qian Yang Yu Yin granules+Benazepril group had a significantly lower urinary microalbuminuria and urinary NAG, there was statistically significant (P<0.01or P<0.05), which the effect of lowering urinary microalbuminuria and urinary NAG is especially combination group significantly (P<0.01).3.Than WKY group, SHR model group obviously increased lesion integral of kidney, there is statistical significance (P＜0.01); With SHR group than in Qian Yang Yu Yin granules groups,Benazepril group and Qian Yang Yu Yin granules+Benazepril group had a significantly lower lesion integral of kidney, there was statistically significant (P<0.01or P<0.05), which the effect of lowering lesion integral of kidney is especially combination group significantly (P＜0.01).4.Than WKY group, by HE dyed SHR model group obviously increased fibrosis index of kidney(FI), there is statistical significance (P<0.01); With SHR group than in Qian Yang Yu Yin granules groups,Benazepril group and Qian Yang Yu Yin granules+Benazepril group had a significantly lower FI, there was statistically significant (P<0.01or P<0.05), which the effect of lowering FI is especially combination group significantly (P<0.01).5.Than WKY group, by masson staining SHR model group obviously increased fibrosis index of kidney(FI), there is statistical significance (P<0.01); With SHR group than in Qian Yang Yu Yin granules groups,Benazepril group and Qian Yang Yu Yin granules+Benazepril group had a significantly lower FI, there was statistically significant (P<0.01or P<0.05), which the effect of lowering FI is especially combination group significantly (P<0.01).6.Using immunofluorescence test, WKY group of normal kidney tissue expression of TGF-betal lessened; and than WKY group,SHR group of kidney tissue expression of TGF-betal display area increased obviously, strength significantly increased, significant differences; With SHR group than in With SHR group than in Qian Yang Yu Yin granules groups,Benazepril group and Qian Yang Yu Yin granules+Benazepril group that split the expression of TGF-betal kidney tissue decrease, there is statistical significance, especially the combination group.7.Than with WKY group, TGF-betal mRNA gene expression of SHR increased obviously (P<0.01), significant differences;With the model group,TGF-betal mRNA gene expression of Qian Yang Yu Yin granules,Benazepril group and Qian Yang Yu Yin granules+Benazepril group dropped significantly,there is statistical significance (P<0.01or P<0.05),especially the combination group(P<0.01). Conclusion:Qian Yang Yu Yin granules can effectively reduce blood pressure, and delay the process with kidney injury, and with the combination therapy of western medicine curative effect is stable, is superior to the simple use of traditional Chinese medicine or western medicine; TGF-betal-TIMPs-Col-IV pathway is likely to be one of the control targets of hypertensive renal impairment.