摘要
目的基于TGF-β1/Smad3信号通路探讨益气养阴活血通络方对糖尿病模型大鼠心肌纤维化的影响。方法将40只雄性SD大鼠按随机数字表法分为空白组10只,糖尿病模型组30只。采用高糖高脂饲料喂养联合小剂量腹腔注射STZ方法制备糖尿病大鼠模型。将造模成功的大鼠随机分为模型组10只、中药组9只、西药组9只。西药组灌胃盐酸曲美他嗪片4.16 mg/kg,中药组灌胃益气养阴活血通络方19.1 mg/kg,空白组、模型组灌胃等体积的生理盐水,1次/d,连续灌胃20周。采用血糖仪检测大鼠空腹血糖,采用碱水解法检测血清羟脯氨酸(hydroxyproline, Hyp)水平,采用ELISA法检测血清TGF-β1水平;采用Masson染色观察各组大鼠心肌纤维化程度;采用Western blot法检测心肌组织TGF-β1、Smad3、胶原蛋白Ⅰ(collagen Ⅰ, Col-Ⅰ)、胶原蛋白Ⅲ(collagen Ⅲ, Col-Ⅲ)蛋白表达;采用实时定量PCR法检测TGF-β1、Smad3基因表达。结果与模型组比较,中药组空腹血糖[(19.41±3.65)mmol/L比(27.48±2.36)mmol/L]、血清Hyp[(25.76±1.73)μg/ml比(31.71±0.33)μg/ml]、TGF-β1[(28.41±0.05)pg/ml比(30.82±0.84)pg/ml]水平均明显降低(P<0.05),心肌间质胶原纤维减少;心肌组织TGF-β1蛋白[(0.81±0.02)比(0.96±0.08)]、Smad3蛋白[(0.36±0.05)比(0.57±0.06)]、Col-Ⅰ蛋白[(2.36±0.45)比(3.53±0.58)]、Col-Ⅲ蛋白[(1.92±0.31)比(2.75±0.08)]、TGF-β1 mRNA[(3.55±0.82)比(6.67±1.33)]和Smad3 mRNA[(1.62±0.35)比(2.70±0.85)]表达降低(P<0.05)。结论益气养阴活血通络方可有效缓解心肌纤维化,其作用机制可能与调控TGF-β1/Smad3信号通路有关。
Objective Based on TGF-β1/Smad3 signal pathway,study the effect of Yiqi-Yangyin-Huoxue-Tongluo Recipe in diabetic rat model.Methods Forty male SD rats were randomly divided into blank group(n=10)and diabetic model group(n=30).Diabetic rats were fed with high sugar and high-fat diet combined with low-dose intraperitoneal injection of STZ.The successful rats were randomly divided into model group(n=10),traditional Chinese medicine(TCM)group(n=9)and western medicine group(n=9).The western medicine group was given trimetazidine hydrochloride tablets 4.16 mg/kg,the TCM group was given Yiqi-Yangyin-Huoxue-Tongluo Recipe 19.1 mg/kg,and the blank group and model group were given the same volume of normal saline,once a day for 20 weeks.The level of fasting blood glucose(FBG)in rats were detected by blood glucose meter;Alkaline hydrolysis was used to detect serum hydroxyproline(Hyp);Enzyme-linked immunosorbent assay(ELISA)to detect serum TGF-β1;Masson staining to observe myocardial fibrosis;Western blotting was used to detect TGF-β1,Smad3,collagenⅠ(Col-Ⅰ)and collagenⅢ(Col-Ⅲ)protein expression.The mRNA expression of TGF-β1 and Smad3 were detected by real-time quantitative PCR.Results Compared with the model group,the FBG(19.41±3.65 mmol/L vs.27.48±2.36 mmol/L),serum Hyp(25.76±1.73μg/ml vs.31.71±0.33μg/ml),TGF-β1(28.41±0.05 pg/ml vs.30.82±0.84 pg/ml)of TCM group were significantly reduced(P<0.05),and myocardial interstitial collagen fibers decreased.The expression of TGF-β1(0.81±0.02 vs.0.96±0.08),Smad3(0.36±0.05 vs.0.57±0.06),Col-Ⅰ(2.36±0.45 vs.3.53±0.58),Col-Ⅲ(1.92±0.31 vs.2.75±0.08)protein in the myocardial tissue of TCM group significantly decreased(P<0.05),the mRNA expression of TGF-β1(3.55±0.82 vs.6.67±1.33)and Smad3(1.62±0.35 vs.2.70±0.85)in myocardial tissue significantly decreased(P<0.05).Conclusions Yiqi-Yangyin-Huoxue-Tongluo Recipe can effectively treat myocardial fibrosis,and its mechanism may be related to the regulation of TGF-β1/Smad3 signal pathway.
作者
吴媛
张贺芳
牛明明
Wu Yuan;Zhang Hefang;Niu Mingming(Graduate School,Hebei University of Chinese Medicine,Shijiazhuang 050091,China;Department of Endocrinology,Hebei Hospital of Traditional ChineseMedicine,Affiliated Hospital of Hebei University of Traditional Chinese Medicine,Shijiazhuang 050011,China)
出处
《国际中医中药杂志》
2020年第11期1088-1093,共6页
International Journal of Traditional Chinese Medicine
基金
河北省科技厅科技支撑计划项目(17277725D)。
