玉竹提取物对心肌缺血再灌注损伤大鼠心肌细胞线粒体损伤及凋亡的保护作用

杨禹晗; 陈超英; 袁占鹏

doi:10.13422/j.cnki.syfjx.20181626

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玉竹提取物对心肌缺血再灌注损伤大鼠心肌细胞线粒体损伤及凋亡的保护作用

药理 | 更新时间：2024-01-04

- 玉竹提取物对心肌缺血再灌注损伤大鼠心肌细胞线粒体损伤及凋亡的保护作用
- Protective Effect of Ploygonati Odorati Rhizoma Extract Against Mitochondrial Damage and Apoptosis in Myocardial Cells of Myocardial Ischemia-reperfusion Injury in Rats
- 中国实验方剂学杂志 2018年24卷第16期页码：136-140
- 作者机构：
- 作者简介：
- 基金信息：
  
  应用毒理湖北省重点实验室项目（201420002）
- DOI：10.13422/j.cnki.syfjx.20181626
  中图分类号： R285.5
- 纸质出版日期：2018
- 稿件说明：
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杨禹晗, 陈超英, 袁占鹏. 玉竹提取物对心肌缺血再灌注损伤大鼠心肌细胞线粒体损伤及凋亡的保护作用[J]. 中国实验方剂学杂志, 2018,24(16):136-140.

YANG Yu-han, CHEN Chao-ying, YUAN Zhan-peng. Protective Effect of Ploygonati Odorati Rhizoma Extract Against Mitochondrial Damage and Apoptosis in Myocardial Cells of Myocardial Ischemia-reperfusion Injury in Rats[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(16): 136-140.
杨禹晗, 陈超英, 袁占鹏. 玉竹提取物对心肌缺血再灌注损伤大鼠心肌细胞线粒体损伤及凋亡的保护作用[J]. 中国实验方剂学杂志, 2018,24(16):136-140. DOI： 10.13422/j.cnki.syfjx.20181626.

YANG Yu-han, CHEN Chao-ying, YUAN Zhan-peng. Protective Effect of Ploygonati Odorati Rhizoma Extract Against Mitochondrial Damage and Apoptosis in Myocardial Cells of Myocardial Ischemia-reperfusion Injury in Rats[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(16): 136-140. DOI： 10.13422/j.cnki.syfjx.20181626.

摘要

目的：探究玉竹提取物对心肌缺血再灌注损伤大鼠心肌细胞线粒体损伤及凋亡的保护作用。方法：选取雄性Wistar大鼠60只，随机分为假手术组、模型组、玉竹提取物低、中、高剂量组及银杏内酯B组，采用可逆性左前降支结扎法建立心肌缺血再灌注模型，给予大鼠心肌缺血30 min，再灌注120 min。玉竹提取物低、中、高剂量组及银杏内酯B组在缺血前30 min给予玉竹提取物100，200，300 mg·kg-1及银杏内酯B 60 mg·kg-1腹腔注射，假手术组及模型组给予相同体积0.9% NaCl溶液腹腔注射；记录大鼠血流动力学指标；采用原位末端转移酶标记法（TUNEL）染色观察各组大鼠心室肌细胞凋亡情况；采用透射电子显微镜观察大鼠心肌线粒体形态及病理变化，采用酶联免疫吸附法（ELISA）检测大鼠心肌组织游离脂肪酸（FFA），三磷酸腺苷（ATP），腺苷酸（AMP），乳酸（LAC）含量；采用蛋白免疫印迹法（Western blot）检测各组大鼠心室肌组织半胱氨酸蛋白酶（Caspase）-3，Caspase-9，Caspase-12蛋白的表达。分离心肌线粒体检测其超氧化物歧化酶（SOD）活性及丙二醛（MDA）及游离钙含量。结果：与假手术组比较，模型组大鼠左室收缩最大压、左心室压力最大上升速率下降，左室舒张末压、左心室压力最大下降速率升高，心肌细胞凋亡明显，心肌组织FFA，LAC含量升高，Caspase-3，Caspase-9，Caspase-12蛋白表达明显增加，ATP/AMP值明显降低，线粒体膜崩解、内嵴明显消失（P<0.05）；与模型组比较，玉竹提取物低、中、高剂量及银杏内酯B组大鼠血流动力学指标、心肌细胞凋亡程度及心肌线粒体病理变化均有改善，心肌组织FFA，LAC含量明显降低，Caspase-3，Caspase-9，Caspase-12蛋白表达降低，ATP/AMP值明显升高（P<0.05）。结论：玉竹提取物可改善大鼠心肌缺血再灌注损伤，减少心肌细胞凋亡，其机制可能与保护线粒体，增强心肌细胞线粒体能量代谢有关。

Abstract

Objective: To investigate the protective effect of Ploygonati Odorati Rhizoma (PORE) extract against mitochondrial damage and apoptosis in myocardial cells of myocardial ischemia-reperfusion injury in rats. Method: Sixty male Wistar rats were randomly divided into sham operation group

model group

PORE group and ginkgolide B group. Myocardial ischemia-reperfusion model was established through the reversible left anterior descending coronary artery ligation

and the rats were provided with the myocardial ischemia for 30 min and the reperfusion for 120 min. 30 min before ligation

the PORE group was given 100

200

300 mg·kg-1 ip PORE

and the ginkgolide B group were given 60 mg·kg-1 ip

the model group and the sham operation group were given the same volume of saline ip; Hemodynamic indexes of the rats were record. The apoptosis of ventricular myocytes in each group was observed by terminal-deoxynucleoitidyl transferose mediated nick end labeling(TUNEL) staining. The morphology and pathological changes of myocardial mitochondria were observed by transmission electron microscopy. The levels of free fatty acid (FFA)

adenosine monophosphate (ATP)

adenosine monophosphate (AMP)

lactate (LAC) in myocardium of the rats were detected by enzyme-linked immunosorbent assay (ELISA). The protein expressions of cysteine aspartate-specific protease(Caspase)-3

9 and 12 in ventricular myocytes were detected by Western blot. Myocardial mitochondria were isolated for the determination of free calcium content. Result: Compared with the sham operation group

the left ventricular systolic pressure and the maximal rising rate of left ventricular pressure were decreased in the model group; the left ventricular end diastolic pressure

and the maximum descending rate of left ventricular pressure were increased. Myocardial apoptosis was obvious. The contents of FFA and LAC in myocardium were increased

the expressions of Caspase-3

9 and 12 were increased

the ratio of ATP/AMP was decreased

the mitochondrial membrane was disintegrated

and the inner crest was disappeared; compared with the model group

the hemodynamic parameters

myocardial cell apoptosis and myocardial mitochondrial pathological changes of the PORE groups and the ginkgolide B group were improved

the levels of FFA and LAC in myocardial tissue and the levels of Caspase-3

12 were decreased

and the ratio of ATP/AMP was increased (P<0.05). Conclusion: PORE can improve myocardial ischemia-reperfusion injury and reduce myocardial apoptosis

the mechanism may be related to the protection of mitochondria and the enhancement of myocardial cell mitochondrial energy metabolism.

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