丹酚酸B对脓毒症大鼠心肌损伤的保护作用

刘晓龙; 崔子林; 李阳; 张雅敏

doi:10.13422/j.cnki.syfjx.20191337

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丹酚酸B对脓毒症大鼠心肌损伤的保护作用

药理 | 更新时间：2021-02-09

- 丹酚酸B对脓毒症大鼠心肌损伤的保护作用
- Protective Effect of Salvianolic Acid B on Myocardial Injury in Septic Rats
- 中国实验方剂学杂志 2019年25卷第14期页码：112-118
- 作者机构：
  
  天津市第一中心医院，天津　 300192
- 作者简介：
  
  刘晓龙，硕士，主治医师，从事肝胆外科及心血管研究，E-mail：3166425349@qq.com
  张雅敏，博士，主任医师，从事肝再生与保护研究，E-mail：zhangyamin@126.com
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191337
  中图分类号： R285.5; R541; R392.3
- 收稿日期：2018-12-08，
  
  网络出版日期：2019-03-19，
  
  纸质出版日期：2019-07-20
- 稿件说明：
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刘晓龙, 崔子林, 李阳, 等. 丹酚酸B对脓毒症大鼠心肌损伤的保护作用[J]. 中国实验方剂学杂志, 2019,25(14):112-118.

Xiao-long LIU, Zi-lin CUI, Yang LI, et al. Protective Effect of Salvianolic Acid B on Myocardial Injury in Septic Rats[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 112-118.
刘晓龙, 崔子林, 李阳, 等. 丹酚酸B对脓毒症大鼠心肌损伤的保护作用[J]. 中国实验方剂学杂志, 2019,25(14):112-118. DOI： 10.13422/j.cnki.syfjx.20191337.

Xiao-long LIU, Zi-lin CUI, Yang LI, et al. Protective Effect of Salvianolic Acid B on Myocardial Injury in Septic Rats[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 112-118. DOI： 10.13422/j.cnki.syfjx.20191337.

摘要

目的：

探讨丹酚酸B(salvianolic acid B，Sal-B)对脓毒症大鼠心肌损伤的作用及机制。

方法：

将50只雄性SD大鼠随机分为假手术组，模型组(行盲肠结扎穿孔术复制脓毒症动物模型)，Sal-B组低、中、高剂量组(在模型基础上，经尾静脉注射Sal-B 6，12，24 mg·kg

－1

)。采用酶联免疫吸附测定(ELISA)检测心肌坏死标志物和炎症因子，包括血清肌钙蛋白T(TnT)，肌酸激酶同工酶(CK-MB)，心肌肌钙蛋白I(cTnI)和肿瘤坏死因子-

(TNF-

)，白细胞介素(IL)-1

，IL-6水平；比色法检测心肌中超氧化物歧化酶(SOD)的活性和丙二醛(MDA)的含量；采用苏木素-伊红(HE)染色观察各组心肌组织病理变化；采用原位末端标记法(TUNEL)检测心肌细胞凋亡情况；采用蛋白免疫印迹法(Western blot)检测心肌组织中凋亡相关蛋白半胱氨酸蛋白酶-3(Caspase-3)，B淋巴细胞瘤-2(Bcl-2)，Bcl-2相关X蛋白(Bax)和自噬微管相关蛋白轻链3(LC3)，Beclin-1表达水平。

结果：

与假手术组比较，模型组TnT，CK-MB和cTnI水平显著升高(

0.01)；心肌组织TNF-

，IL-1

，IL-6水平显著升高(

0.01)；心肌组织MDA含量显著升高(

0.01)，SOD的活力显著降低(

0.01)，TUNEL阳性心肌细胞比率显著升高(

0.01)，Bax和Caspase-3蛋白表达水平升高，而Bcl-2表达显著降低(

0.01)，自噬相关蛋白LC3，Beclin-1表达水平显著升高(

0.01)；与模型组比较，Sal-B中、高剂量组TnT，CK-MB和cTnI水平均显著降低(

0.01)，心肌组织TNF-

，IL-1

，IL-6水平显著降低(

0.01)，心肌组织MDA含量明显降低(

0.05，

0.01)，同时，心肌组织SOD的活性明显升高(

0.05，

0.01)；TUNEL阳性心肌细胞比率明显降低(

0.05)，Bax和Caspase-3蛋白表达下降，Bcl-2表达明显升高(

0.05，

0.01)；同时增强自噬反应，LC3Ⅱ/LC3Ⅰ增加，Beclin-1表达水平明显上调(

0.05，

0.01)。

结论：

Sal-B通过影响自噬蛋白，降低脓毒症大鼠心肌炎症反应和氧化应激，抑制心肌细胞凋亡，从而减轻脓毒症大鼠心肌的损伤。

Abstract

Objective:

To explore the effect of salvianolic acid B (Sal-B) on the myocardial apoptosis in rats with sepsis.

Method:

The 50 male SD rats were randomly divided into sham operation group

model group (cecal ligation and perforation were performed to replicate the animal model of sepsis)and Sal-B low

medium and high-dose group(6

24 mg·kg

-1

). The myocardial necrosis markers troponin T(TnT)

cardiac troponin I (cTnI)

creatine kinase-MB (CK-MB) and the tumor necrosis factor-

(TNF-

)

interleukin-1

(IL-1

) and interleukin-6 (IL-6) were assayed by enzyme-linked immunosorbent assay(ELISA). The antioxidant enzyme superoxide dismutase (SOD) activities and malonaldialdehyde (MDA) levels in myocardial homogenates were determined by spectrophotometrically. Haematoxylin-eosin (HE) staining was used to evaluate the pathological change in rats. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) was used to detect apoptosis of myocardial cells in hearts

respectively. Western blot was used to detect the expression levels of apoptosis-related protein cysteine protease-3 (Caspase-3)

b-lymphocytoma-2 (Bcl-2)

Bcl-2-related X protein (Bax)

autophagy microtubule-related light chain 3 (LC3) and Beclin-1 in myocardial tissue.

Result:

Compared with sham group

the levels of TnT

CK-MB and cTnI in model group were significantly increased (

0.01). The levels of TNF-

IL-1

and IL-6 in myocardial tissue were significantly increased (

0.01). MDA content in myocardial tissue was significantly increased (

0.01)

SOD activity in myocardial tissue was significantly decreased (

0.01)

the ratio of TUNEL positive myocardial cells was significantly increased (

0.01)

Bax and Caspase-3 protein expression levels were significantly increased

while Bcl-2 expression was significantly decreased (

0.01)

autophagy related protein LC3 and Beclin-1 expression levels were significantly increased (

0.01). Compared with model group

the levels of TnT

CK-MB and cTnI in Sal-B medium and high-dose groups were significantly decreased (

0.01)

the levels of TNF-

IL-1 and IL-6 in the myocardial tissue were significantly decreased (

0.01)

MDA content in myocardial tissue was significantly decreased (

0.05

0.01)

and the SOD activity in myocardial tissue was significantly increased (

0.05

0.01). The ratio of TUNEL positive cardiomyocytes was significantly decreased (

0.05)

Bax and Caspase-3 protein expressions were decreased

and Bcl-2 expression was significantly increased (

0.05

0.01). The autophagy response was enhanced

the LC3Ⅱ/LC3Ⅰ ratio was increased

and Beclin-1 expression was significantly up-regulated (

0.05

0.01).

Conclusion:

Sal-B showed a protective effect on the myocardial apoptosis in septic rats maybe by depressing inflammatory infiltration

anti-oxidative effects and improving the exceptional expression of the proteins such as Bax

Bcl-2

Caspases-3 by enhancing the level of autophagy.

关键词

Keywords

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