Interventive Effect of Zhuangtongyin on Expression of AngⅡ, AT-1R and Cx43 in Rats with MIRI

HUANG Xiao-shan; ZHANG Shi-tian; TANG Han-qing; HUANG Cen-han; LIU Yan-ping; LAN Huang-qi; ZHAO Qiu-hua; LI Ke-ming

doi:10.13422/j.cnki.syfjx.20181053

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Interventive Effect of Zhuangtongyin on Expression of AngⅡ, AT-1R and Cx43 in Rats with MIRI

更新时间：2024-01-04

- Interventive Effect of Zhuangtongyin on Expression of AngⅡ, AT-1R and Cx43 in Rats with MIRI
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 16, Pages: 157-162(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20181053
  CLC： R285.5
- Published：2018
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HUANG Xiao-shan, ZHANG Shi-tian, TANG Han-qing, et al. Interventive Effect of Zhuangtongyin on Expression of AngⅡ, AT-1R and Cx43 in Rats with MIRI[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(16): 157-162.
DOI：

HUANG Xiao-shan, ZHANG Shi-tian, TANG Han-qing, et al. Interventive Effect of Zhuangtongyin on Expression of AngⅡ, AT-1R and Cx43 in Rats with MIRI[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(16): 157-162. DOI： 10.13422/j.cnki.syfjx.20181053.

摘要

目的：观察壮通饮（Zhuangtongyin，ZTY）对心肌缺血再灌注大鼠心肌细胞的血管紧张素Ⅱ（AngⅡ），血管紧张素Ⅱ 1型受体（AT-1R），缝隙连接蛋白43（Cx43） mRNA表达水平和蛋白水平的影响，并探讨ZTY预处理在大鼠心肌缺血再灌注损伤过程中对大鼠心肌细胞的保护机制。方法：采用随机分组的方法将SPF级SD大鼠分为空白组，假手术组（只穿线，不结扎），模型组，壮通饮低、中、高剂量组（6.8，13.6，27.2 g·kg-1）和复方丹参组（0.08 g·kg-1）。连续灌药4周后，通过结扎左冠状动脉前降支30 min，再灌注60 min，建立冠心病血瘀证再灌注损伤模型，术中监测大鼠心电指标，记录心律失常发生情况，再灌注结束后，取下心脏组织，用实时荧光定量PCR（Real-time PCR）检测大鼠心肌组织中AngⅡ，AT-1R，Cx43 mRNA的表达水平，用蛋白免疫印迹法（Western blot）检测大鼠心肌组织中AT-1R，Cx43的蛋白表达水平。结果：壮通饮各剂量组和模型组比较均可以减少心肌缺血再灌注损伤模型大鼠心律失常的发生（P<0.05）。与空白组、假手术组比较，模型组、壮通饮各剂量组、复方丹参组的AngⅡ，AT-1R mRNA表达均上调，Cx43 mRNA表达均下降（P<0.05）。与模型组比较，壮通饮各剂量组、复方丹参组的AngⅡ，AT-1R mRNA表达均下降，Cx43 mRNA表达均上升，各剂量组之间，中剂量组AngⅡ，AT-1R mRNA下降，Cx43 mRNA表达上升最明显（P<0.05）。壮通饮各剂量组与复方丹参组比较无统计学意义。空白组与假手术组比较，无显著性差异。与空白组、假手术比较，模型组、壮通饮各剂量组、复方丹参组的AT-1R蛋白表达均上调，Cx43蛋白表达均下降（P<0.05）。与模型组比较，壮通饮各剂量组、复方丹参组的AT-1R蛋白表达均下降，Cx43蛋白表达均上升，各剂量组之间，中剂量组AT-1R蛋白下降，Cx43蛋白表达上升最明显（P<0.05）。壮通饮各剂量组与复方丹参组对比无统计学意义。结论： ZTY对心肌缺血再灌注损伤有保护作用，降低再灌注心律失常的发生，其作用机制可能与壮通饮能调控AngⅡ-（AT-1R）-Cx43通路，导致Cx43表达上调有关。

Abstract

Objective: To investigate the effects of Zhuangtongyin(ZTY)on the mRNA expression of angiotensin Ⅱ (AngⅡ)

angiotensin-receptor 1 (AT-1R) and connexin 43 (Cx43) in rats with myocardial ischemia-reperfusion injury (MIRI)

and explore the protective mechanism of ZTY pre-treatment on cardiac myocytes in rats during MIRI process. Method: SPF grade SD rats were randomly divided into normal group

sham-operated group (thread without ligation)

model group

ZTY low dose group (6.8 g·kg-1)

ZTY middle dose group (13.6 g·kg-1)

ZTY high dose group (27.2 g·kg-1) and compound Danshen group (0.08 g·kg-1). After drug treatment for continuous 4 weeks

the left anterior descending coronary artery was ligated for 30 min

and then reperfused for 60 min to establish reperfusion injury model in blood stasis syndrome type coronary heart disease. ECG monitoring was performed during operation and the occurrence of arrhythmia was recorded. After reperfusion

their heart tissues were taken and the Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the AngⅡ

AT-1R and Cx43 mRNA expression in myocardial tissues; Western blot method was used to detect the AT-1R and Cx43 protein expression in myocardial tissues. Result: ZTY low dose group

middle dose group and high dose group can reduce the incidence of arrhythmia in rats with myocardial ischemia reperfusion as compared with model group (P<0.05). As compared with the normal group and sham operation group

the AngⅡand AT-1R mRNA expression levels were increased significantly

and Cx43 expression levels were reduced in model group

all ZTY groups and compound Danshen group (P<0.05). As compared with the model group

the AngⅡand AT-1R mRNA levels were reduced and Cx43 mRNA expression levels were increased in all ZTY groups and compound Danshen group (P<0.05)

and the effect was most significant in ZTY middle dose group (P<0.05). There was no significant difference between ZTY groups and compound Danshen group in AngⅡ

AT-1R and Cx43.There was no significant difference between normal group and sham operation group.Compared with normal group

the myocardial ischemia-reperfusion injury rats

ZTY low

middle

high dose group and compound Salvia Pellet dose group myocardial organization AT-1R is increased sinificantly(P<0.05)

Cx43 reduced(P<0.05). Compared with ischemia-reperfusion injury rats

ZTY low

middle

high dose group and compound Salvia Pellet dose group myocardial organization AT-1R is reduced sinificantly(P<0.05)

Cx43 increased (P<0.05)

ZTY middle dose group better than other group (P<0.05).There was no sinificant difference between ZTY each dose group and compound Salvia Pellet dose group in AT-1R and Cx43. Conclusion: ZTY can reduce myocardial ischemia-reperfusion arrhythmia and have protective effects on myocardial ischemia-reperfusion injury

and the mechanism may be associated with regulating AngⅡ-(AT-1R)-Cx43 pathway and up-regulating Cx43 expression in myocardial tissues.

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