调脾护心方对急性心肌缺血模型大鼠心肌AngⅡ、ACE、AGTR1蛋白及AGT、AGTR1、ET-1 mRNA表达的影响
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摘要
目的观察调脾护心方对急性心肌缺血模型大鼠心肌血管紧张素Ⅱ(AngⅡ)、血管紧张素转化酶(ACE)、血管紧张素Ⅱ受体1(AGTR1)蛋白和血管紧张素原(AGT)、AGTR1、内皮缩血管肽1(ET-1)mRNA表达的影响。方法采用腹腔注射异丙肾上腺素[5 mg/(kg·d)]连续5天,制备大鼠急性心肌缺血模型。将64只急性心肌缺血模型大鼠随机分为调脾护心方低剂量组[低剂量组,生药5.85 mg/(kg·d)]、调脾护心方高剂量组[高剂量组,生药23.40 mg/(kg·d)]、曲美他嗪组[10 mg/(kg·d)]、模型组,每组16只,并设正常大鼠16只作为对照组。各用药组灌胃相应药物,对照组与模型组灌胃等体积生理盐水。连续灌胃28天后,分别采用HE染色法观察心肌组织病理变化;Western Blot检测各组大鼠心肌组织中AngⅡ、ACE、AGTR1蛋白表达;RT-PCR检测各组大鼠心肌组织中AGT、AGTR1、ET-1 mRNA表达。结果病理结果显示,调脾护心方和曲美他嗪可改善心肌缺血后细胞损伤。与对照组比较,模型组AngⅡ、ACE、AGTR1蛋白和AGT、AGTR1、 ET-1 mRNA表达增高(P<0.05,P<0.01)。与模型组比较,各用药组AngⅡ、ACE、AGTR1蛋白和AGT、AGTR1、 ET-1 mRNA表达均降低(P<0.05,P<0.01)。与曲美他嗪组比较,低剂量组AngⅡ、ACE、AGTR1蛋白和AGT、AGTR1 mRNA表达增高,高剂量组ACE蛋白和AGT mRNA表达增高(P<0.05,P<0.01)。与低剂量组比较,高剂量组AngⅡ、ACE、AGTR1蛋白和AGT mRNA表达降低(P<0.01),ET-1 mRNA表达增高(P<0.01)。结论调脾护心方可通过抑制肾素—血管紧张素系统(RAS)的过度激活发挥心肌保护作用。
Objective To observe the effects of Tiaopi Huxin Decoction(TPHXD) on the protein expressions of angiotensin Ⅱ(Ang Ⅱ), angiotensin converting enzyme(ACE), angiotensin Ⅱ receptor 1(AGTR1) and mRNA expressions of angiotensinogen(AGT), AGTR1 and endothelial vasoconstrictor peptide 1(ET-1) in acute myocardial ischemia model rats. Methods Isoprenaline(5 mg·kg~(-1)·d~(-1)) were given to rats for 5 consecutive days to establish acute myocardial ischemia model. Totally 64 acute myocardial ischemia model rats were randomly divided into TPHXD low-dose group(5.85 mg·kg~(-1)·d~(-1)), TPHXD high-dose group(23.4 mg·kg~(-1)·d~(-1)), trimetazidine group(10 mg·kg~(-1)·d~(-1)), and model group, 16 rats in each group. Another 16 normal rats were selected as control group. The intervention groups were given TPHXD or trimetazidine respectively. The control group and model group were given equal volume of normal saline. After 28 days of gavage, the pathological changes of myocardial tissue were observed by HE staining. The protein expression of AngⅡ, ACE and AGTR1 in the myocardium of each group was detected by Western Blot. The AGT, AGTR1 and ET-1 mRNA expression in each group were detected by Real-time PCR. Results Pathological results showed that TPHXD and trimetazidine could alleviate the cell injury by myocardial ischemia. Compared with the control group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT, AGTR1, ET-1 mRNA increased in the model group(P<0.05, P<0.01). Compared with the model group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT, AGTR1 mRNA decreased in intervention groups(P<0.05, P<0.01). Compared with the trimetazidine group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT, AGTR1 mRNA increased in TPHXD low-dose group, while the expressions of ACE protein and AGT mRNA increased in TPHXD high-dose group(P<0.05, P<0.01); compared with the TPHXD low-dose group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT mRNA decreased in TPHXD high-dose group(P<0.01), mRNA expression of ET-1 was increased. Conclusion TPHXD can protect damaged myocardium by blocking excessive activation of renin-angiotensin system.
Objective To observe the effects of Tiaopi Huxin Decoction(TPHXD) on the protein expressions of angiotensin Ⅱ(Ang Ⅱ), angiotensin converting enzyme(ACE), angiotensin Ⅱ receptor 1(AGTR1) and mRNA expressions of angiotensinogen(AGT), AGTR1 and endothelial vasoconstrictor peptide 1(ET-1) in acute myocardial ischemia model rats. Methods Isoprenaline(5 mg·kg~(-1)·d~(-1)) were given to rats for 5 consecutive days to establish acute myocardial ischemia model. Totally 64 acute myocardial ischemia model rats were randomly divided into TPHXD low-dose group(5.85 mg·kg~(-1)·d~(-1)), TPHXD high-dose group(23.4 mg·kg~(-1)·d~(-1)), trimetazidine group(10 mg·kg~(-1)·d~(-1)), and model group, 16 rats in each group. Another 16 normal rats were selected as control group. The intervention groups were given TPHXD or trimetazidine respectively. The control group and model group were given equal volume of normal saline. After 28 days of gavage, the pathological changes of myocardial tissue were observed by HE staining. The protein expression of AngⅡ, ACE and AGTR1 in the myocardium of each group was detected by Western Blot. The AGT, AGTR1 and ET-1 mRNA expression in each group were detected by Real-time PCR. Results Pathological results showed that TPHXD and trimetazidine could alleviate the cell injury by myocardial ischemia. Compared with the control group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT, AGTR1, ET-1 mRNA increased in the model group(P<0.05, P<0.01). Compared with the model group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT, AGTR1 mRNA decreased in intervention groups(P<0.05, P<0.01). Compared with the trimetazidine group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT, AGTR1 mRNA increased in TPHXD low-dose group, while the expressions of ACE protein and AGT mRNA increased in TPHXD high-dose group(P<0.05, P<0.01); compared with the TPHXD low-dose group, the expressions of Ang Ⅱ, ACE, AGTR1 protein and AGT mRNA decreased in TPHXD high-dose group(P<0.01), mRNA expression of ET-1 was increased. Conclusion TPHXD can protect damaged myocardium by blocking excessive activation of renin-angiotensin system.
引文
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[2] 杨帆,戴小华,付军,等.加载调脾护心方治疗慢性心力衰竭心脾两虚证临床研究[J].中西医结合心脑血管病杂志,2017,15(8):956-958.
[3] 王越.调脾护心方对急性心肌梗死的临床疗效观察及动物实验研究[D].合肥:安徽中医药大学,2016.
[4] 李真真,徐志懿,关新强.黄芪甲苷对异丙肾上腺素诱导心肌梗死保护作用[J].安徽医科大学学报,2013,48(3):275-278.
[5] 郁秀娟,丁晓梅.心肌梗死后心室重构研究进展[J].中华临床医师杂志(电子版),2013,7(2):723-726.
[6] Raizada V,Skipper B,Luo W,et al.Intracardiac and intrarenal renin-angiotensin systems:mechanisms of cardiovascular and renal effects[J].J Invest Med,2007,55(7):341-59.
[7] 李敏,李虎,王逵,等.硝普钠联合硝酸甘油治疗急性心力衰竭的疗效及对血浆Ang-Ⅱ,NO,ET-1,Cys-C的影响[J].中西医结合心脑血管病杂志,2018,16(3):330-333.
[8] 谢永花.蛭龙活血通瘀胶囊改善大鼠心肌梗死后心室重构机制的实验研究[D].四川:西南医科大学,2016.
[9] Fraga-Silva RA,Da Silva DG,Montecucco F,et al.The angiotensin-converting enzyme 2/angiotensin-(1-7)/Mas receptor axis:a potential target for treat- ing thrombotic diseases[J].Thromb Haemost,2012,108(6):1089-1096.
[10] 袁良,戴小华.调脾护心法论治胸痹的理论研究[J].江西中医药大学学报,2017,29(4):16-18.
[11] 陈一竹,杨文龙,郭玲玉,等.白术内酯Ⅱ抗血小板作用及对血小板中蛋白激酶B磷酸化水平的影响[J].中国医药导报,2016,13(11):18-21,26.
[12] 耿欣,李廷利.酸枣仁主要化学成分及药理作用研究进展[J].中医药学报,2016,44(5):84-86.
[13] Wei XJ,Hu TJ,Chen JR,et al.Inhibitory effect of carboxymethylpachymaran on cyclophosphamide-induced oxidative stress in mice[J].Int J Biol Macromol,2011,49(4):801-805.
[14] 刘大伟,康利平,马百平.远志化学及药理作用研究进展[J].国际药学研究杂志,2012,39(1):32-36,44.
[15] 李伟伟,张国伟.陈皮黄酮类成分研究进展[J].中国医学创新,2014,11(24):154-156.
[16] 侯鹏飞,陈文星,赵新慧,等.木香挥发性成分气质联用分析及其抑制血小板聚集作用的研究[J].中国实验方剂学杂志,2008,14(7):26-30.
[17] 于立恒.蒲公英药理作用研究进展[J].实用中医药杂志,2012,28(7):617-620.
[18] 祝德伟.调脾护心方防治慢性心力衰竭及对炎症因子影响的临床及实验研究[D].合肥:安徽中医药大学,2018.