铁皮石斛对缺血再灌注后心衰心气虚型大鼠心肌纤维化的抑制作用
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摘要
目的:基于缺血再灌注后心衰心气虚型大鼠模型心动周期、心肌病理切片、心肌半乳糖凝集素3(Gal-3)转化生长因子-β(TGF-β),Smad同源物3重组蛋白(Smad3)蛋白表达情况,探讨铁皮石斛对模型大鼠心肌纤维化的干预效果。方法:采用冠状动脉左前降支进行结扎术建立心衰心气虚型大鼠模型,将大鼠分为正常组,模型组,缬沙坦组(9. 43 mg·kg~(-1)),铁皮石斛组(10 mg·kg~(-1))共4组,每组10只,正常组和模型组给予等体积生理盐水。通过动物高分辨超声仪记录各组大鼠心动周期左室舒张末期内径(LVEDD),左室收缩末期内径(LVESD),左心射血分数(LVEF),左心室短轴缩短率(LVFS)变化;酶联免疫吸附测定(ELISA)试剂盒测定I型胶原羧基端前肽(PICP),Ⅲ型胶原羧基端前肽(PⅢNP);苏木素-伊红(HE)染色观察心肌细胞形态变化,马松(Masson)染色观察心肌纤维组织、胶原组织变化,蛋白免疫印迹法(Western blot)检测4组小鼠心肌半乳糖凝集素3(Gal-3),转化生长因子-β(TGF-β),Smad同源物3重组蛋白(Smad3)蛋白表达情况。结果:与正常组比较,模型组LVEDD,LVEF,LVFS水平明显降低(P <0. 05),LVESD,PICP,PIIINP水平升高(P <0. 05),Gal-3,TGF-β,Smad3蛋白表达水平显著升高(P <0. 01);与模型组比较,缬沙坦组、铁皮石斛组LVEDD,LVEF,LVFS明显升高(P <0. 05),LVESD,PICP,PIIINP水平明显降低(P <0. 05),Gal-3,TGF-β,Smad3蛋白表达水平明显降低(P <0. 05,P <0. 01),且铁皮石斛组低于缬沙坦组(P <0. 05); HE染色结果显示,铁皮石斛组心肌细胞形态明显改善,Masson染色结果显示,铁皮石斛组心肌纤维组织、胶原组织形态明显改善。结论:铁皮石斛可有效抑制缺血再灌注后心衰心气虚症候大鼠心肌纤维化,机制可能为降低Gal-3,TGF-β,Smad3蛋白表达有关。
Objective: To investigate the expressions of cardiac cycle,myocardial pathology,galectin-3(Gal-3),transforming growth factor-β(TGF-β),Smad homologue 3 recombinant protein(Smad3) in rats with heart failure and heart failure after ischemia-reperfusion,and the intervention effect of Dendrobii Officinalis Caulis(DOC) myocardial fibrosis in model rats. Method: A rat model of heart failure and Qi deficiency was established through ligation of the left anterior descending coronary artery. The rats were divided into blank group,model group,valsartan group(9. 43 mg·kg~(-1)) and DOC group(10 mg·kg~(-1)),with 10 in each group. The blank group and the model group were given an equal volume of physiological saline. The changes in left ventricular enddiastolic diameter(LVEDD),left ventricular end-systolic dimension(LVESD),left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS) of the cardiac cycle of rats in each group were recorded by high-resolution ultrasound system. The carboxyterrninal propeptide of type I procollagen(PICP), and carboxyterrninal propeptide of type Ⅲ procollagen(PIIINP) were detected by enzyme-linked immunosorbent assay(ELISA) kit. The morphological changes of myocardial cells were observed by hematoxylin-eosin(HE) staining.The changes of myocardial fiber tissue and collagen were observed by Masson staining. Western blot was used to detect the protein expressions of Gal-3,TGF-β,Smad3. Result: Compared with the blank group,the levels of LVEDD,LVEF,and LVFS were lower in the model group(P < 0. 05),the levels of LVESD,PICP and PIIINP were increased(P < 0. 05),while the expression levels of Gal-3,TGF-β,and Smad3 were decreased(P < 0. 05).Compared with the model group,LVEDD,LVEF,LVFS of the control group and the experimental group were better than the model group(P < 0. 05),and LVESD,PICP,PIIINP levels of the experimental group were lower than the control group(P < 0. 05),LVESD,PPIC,PIIINP were lower than those of the model group(P < 0. 05),expression levels of Gal-3,TGF-β and Smad3 were lower than those in the model group(P < 0. 05),and the experimental group was lower than the control group(P < 0. 05). According to HE staining,the morphological changes in myocardial cells in the experimental group were more significant than that in the control group. Masson staining showed that the morphological changes of myocardial fibrous tissue and collagen tissue in the experimental group were more significant than that in the control group. Conclusion: DOC can effectively inhibit myocardial fibrosis in rats with heart failure and heart Qi deficiency syndrome after ischemia-reperfusion. The mechanism may be correlated with the reduction of the expressions of Gal-3,TGF-β and Smad3.
Objective: To investigate the expressions of cardiac cycle,myocardial pathology,galectin-3(Gal-3),transforming growth factor-β(TGF-β),Smad homologue 3 recombinant protein(Smad3) in rats with heart failure and heart failure after ischemia-reperfusion,and the intervention effect of Dendrobii Officinalis Caulis(DOC) myocardial fibrosis in model rats. Method: A rat model of heart failure and Qi deficiency was established through ligation of the left anterior descending coronary artery. The rats were divided into blank group,model group,valsartan group(9. 43 mg·kg~(-1)) and DOC group(10 mg·kg~(-1)),with 10 in each group. The blank group and the model group were given an equal volume of physiological saline. The changes in left ventricular enddiastolic diameter(LVEDD),left ventricular end-systolic dimension(LVESD),left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS) of the cardiac cycle of rats in each group were recorded by high-resolution ultrasound system. The carboxyterrninal propeptide of type I procollagen(PICP), and carboxyterrninal propeptide of type Ⅲ procollagen(PIIINP) were detected by enzyme-linked immunosorbent assay(ELISA) kit. The morphological changes of myocardial cells were observed by hematoxylin-eosin(HE) staining.The changes of myocardial fiber tissue and collagen were observed by Masson staining. Western blot was used to detect the protein expressions of Gal-3,TGF-β,Smad3. Result: Compared with the blank group,the levels of LVEDD,LVEF,and LVFS were lower in the model group(P < 0. 05),the levels of LVESD,PICP and PIIINP were increased(P < 0. 05),while the expression levels of Gal-3,TGF-β,and Smad3 were decreased(P < 0. 05).Compared with the model group,LVEDD,LVEF,LVFS of the control group and the experimental group were better than the model group(P < 0. 05),and LVESD,PICP,PIIINP levels of the experimental group were lower than the control group(P < 0. 05),LVESD,PPIC,PIIINP were lower than those of the model group(P < 0. 05),expression levels of Gal-3,TGF-β and Smad3 were lower than those in the model group(P < 0. 05),and the experimental group was lower than the control group(P < 0. 05). According to HE staining,the morphological changes in myocardial cells in the experimental group were more significant than that in the control group. Masson staining showed that the morphological changes of myocardial fibrous tissue and collagen tissue in the experimental group were more significant than that in the control group. Conclusion: DOC can effectively inhibit myocardial fibrosis in rats with heart failure and heart Qi deficiency syndrome after ischemia-reperfusion. The mechanism may be correlated with the reduction of the expressions of Gal-3,TGF-β and Smad3.
引文
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[18]翟旭峰,王瑾,郑爽,等.铁皮石斛对缺血再灌注诱导的心律失常的保护作用[J].现代食品科技,2017,33(7):1-8.
[19]赵桂峰,吴丽玉,许传嘉.芪参益气滴丸对心梗后大鼠心功能及心肌纤维化相关蛋白表达的影响[J].中国实验方剂学杂志,2018,24(4):131-136.
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[21]Ravassa S,López B,Querejeta R,et al.Phenotyping of myocardial fibrosis in hypertensive patients with heart failure.Influence on clinical outcome[J].J Hypertens,2017,35(4):853-861.
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[23]黄海烽,王春花,赵玲璐,等.丹酚酸B抑制血管紧张素Ⅱ诱导心肌成纤维细胞增殖与分化的作用[J].中国实验方剂学杂志,2017,23(13):128-132.
[24]肖一佳,沈祥春,李杰平,等.心肌纤维化与TGF-β-Smad信号系统[J].中国新药杂志,2016,25(2):182-186.
[25]Khan S A,DONG H,Jennifer J,et al.Fibulin-2 is essential for angiotensin II-induced myocardial fibrosis mediated by transforming growth factor(TGF)-β[J].Lab Invest,2016,96(7):773-783.
[26]WU N,GE Y,ZHOU Y,et al.Nanosized titanium dioxide resulted in the activation of TGF-β/Smads/p38MAPK pathway in renal inflammation and fibration of mice[J].J Biomed Mater Res A,2016,104(6):1452-1461.
[27]石斌豪,徐宗佩,樊官伟.基于TGF-β1/Smads信号通路治疗心肌梗死后心肌纤维化的研究进展[J].中国药理学通报,2018,34(1):5-8.
[2]马金,张艳.慢性心力衰竭中医病机“气虚-血瘀-水停”与“心室重构”的相关性探讨[J].现代中西医结合杂志,2016,25(17):1936-1938.
[3]孙龙飞,安冬青,郭龙龙.心力衰竭的中医药治疗优势与特色[J].中国中医急症,2016,25(3):452-456.
[4]王怀隐.太平圣惠方[M].北京:人民卫生出版社,1958:245.
[5]陈桦,王兵,唐汉庆,等.铁皮石斛多糖对冠心病模型家兔心功能及心肌收缩能力的影响[J].中国实验方剂学杂志,2015,21(21):139-143.
[6]唐汉庆,赵玉峰,李天资,等.铁皮石斛对冠心病模型家兔心功能和血管变化的影响[J].世界科学技术-中医药现代化,2015,17(4):856-860.
[7]赵智强,吴勉华,周学平,等.周仲瑛教授从瘀热论治心脑血管疾病学术思想探讨[J].辽宁中医药大学学报,2011,13(9):5-7.
[8]黄小珊,张世田,唐汉庆,等.壮通饮预处理对冠心病血瘀证大鼠AngⅡ,AT-1R,Cx43及其对再灌注损伤的影响[J].中国实验方剂学杂志,2018,24(16):157-162.
[9]王伟伟.AngⅡ/AT1R与PI3K/Akt/e NOS信号通路在高血压心房结构重塑中的作用以及替米沙坦对高血压大鼠房颤易感性的影响[D].福州:福建医科大学,2015.
[10]周琪.铁皮石斛水提物对糖尿病小鼠血糖和肠道菌群的影响[D].大连:大连医科大学,2017.
[11]Grassi G,Seravalle G,Quarti-Trevano F,et al.Excessive sympathetic activation in heart failure with obesity and metabolic syndrome:characteristics and mechanisms[J].Hypertension,2016,49(3):535-541.
[12]田鲜美,刘宇,王玫,等.参附注射液治疗急性心力衰竭(心气阳虚证)并快速性心律失常的临床观察[J].中国中医急症,2017,26(4):676-679.
[13]李鹤,刘亚洋,徐素娥,等.益气舒心汤对舒张性心力衰竭(心气虚证)病人心功能及生活质量的影响[J].中西医结合心脑血管病杂志,2016,14(7):693-695.
[14]LIU Y,YANG H,LIU L X,et al.NOD2 contributes to myocardial ischemia/reperfusion injury by regulating cardiomyocyte apoptosis and inflammation[J].Life Sci,2016,149(4):10-16.
[15]张四杰,钱正,刘京晶,等.铁皮石斛花中花色成分抗氧化性和稳定性研究[J].中国中药杂志,2018,43(10):2025-2031.
[16]梁凯伦,方萍,施秋秋,等.铁皮石斛花对高糖高脂饮酒致高血压大鼠的降压作用及机制研究[J].中国中药杂志,2018,43(1):147-153.
[17]唐汉庆,王兵,廉春容,等.铁皮石斛多糖对冠心病缓慢性心律失常大鼠NO、c GMP及Na+-K+-ATP酶活性的影响[J].动物医学进展,2016,37(12):77-81.
[18]翟旭峰,王瑾,郑爽,等.铁皮石斛对缺血再灌注诱导的心律失常的保护作用[J].现代食品科技,2017,33(7):1-8.
[19]赵桂峰,吴丽玉,许传嘉.芪参益气滴丸对心梗后大鼠心功能及心肌纤维化相关蛋白表达的影响[J].中国实验方剂学杂志,2018,24(4):131-136.
[20]MA K,ZHANG C,HUANG M Y,et al.Crosstalk between Beclin-1-dependent autophagy and caspasedependent apoptosis induced by tanshinone IIA in human osteosarcoma MG-63 cells[J].Oncol Rep,2016,36(4):1807-1818.
[21]Ravassa S,López B,Querejeta R,et al.Phenotyping of myocardial fibrosis in hypertensive patients with heart failure.Influence on clinical outcome[J].J Hypertens,2017,35(4):853-861.
[22]Souza D F,Silva D N,Carvalho R H,et al.Association of cardiac galectin-3 expression,myocarditis,and fibrosis in chronic chagas disease cardiomyopathy[J].Am JPathol,2017,187(5):1134-1146.
[23]黄海烽,王春花,赵玲璐,等.丹酚酸B抑制血管紧张素Ⅱ诱导心肌成纤维细胞增殖与分化的作用[J].中国实验方剂学杂志,2017,23(13):128-132.
[24]肖一佳,沈祥春,李杰平,等.心肌纤维化与TGF-β-Smad信号系统[J].中国新药杂志,2016,25(2):182-186.
[25]Khan S A,DONG H,Jennifer J,et al.Fibulin-2 is essential for angiotensin II-induced myocardial fibrosis mediated by transforming growth factor(TGF)-β[J].Lab Invest,2016,96(7):773-783.
[26]WU N,GE Y,ZHOU Y,et al.Nanosized titanium dioxide resulted in the activation of TGF-β/Smads/p38MAPK pathway in renal inflammation and fibration of mice[J].J Biomed Mater Res A,2016,104(6):1452-1461.
[27]石斌豪,徐宗佩,樊官伟.基于TGF-β1/Smads信号通路治疗心肌梗死后心肌纤维化的研究进展[J].中国药理学通报,2018,34(1):5-8.