黄芪甲苷对异丙肾上腺素诱导的大鼠原代心肌细胞损伤的抗凋亡作用

武梦羚; 宋耀鸿

doi:10.13422/j.cnki.syfjx.20181035

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黄芪甲苷对异丙肾上腺素诱导的大鼠原代心肌细胞损伤的抗凋亡作用

药理 | 更新时间：2024-01-04

- 黄芪甲苷对异丙肾上腺素诱导的大鼠原代心肌细胞损伤的抗凋亡作用
- Anti-apoptosis Effect of Astragaloside on Isoproterenol-induced Rat Primary Cardiomyocytes
- 中国实验方剂学杂志 2018年24卷第10期页码：137-143
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81273716）;南京市科技计划项目（201201067）;江苏省“六大人才高峰”项目（2015-WSN-067）
- DOI：10.13422/j.cnki.syfjx.20181035
  中图分类号： R285.5
- 纸质出版日期：2018
- 稿件说明：
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武梦羚, 宋耀鸿. 黄芪甲苷对异丙肾上腺素诱导的大鼠原代心肌细胞损伤的抗凋亡作用[J]. 中国实验方剂学杂志, 2018,24(10):137-143.

WU Meng-ling, SONG Yao-hong. Anti-apoptosis Effect of Astragaloside on Isoproterenol-induced Rat Primary Cardiomyocytes[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 137-143.
武梦羚, 宋耀鸿. 黄芪甲苷对异丙肾上腺素诱导的大鼠原代心肌细胞损伤的抗凋亡作用[J]. 中国实验方剂学杂志, 2018,24(10):137-143. DOI： 10.13422/j.cnki.syfjx.20181035.

WU Meng-ling, SONG Yao-hong. Anti-apoptosis Effect of Astragaloside on Isoproterenol-induced Rat Primary Cardiomyocytes[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 137-143. DOI： 10.13422/j.cnki.syfjx.20181035.

摘要

目的：研究黄芪甲苷对异丙肾上腺素诱导的SD大鼠乳鼠原代心肌细胞损伤的抗凋亡作用，并探讨相关的作用机制。方法：利用酶解法和差速贴壁方法从出生1~3 d的正常SD大鼠的乳鼠心脏中分离得到原代心肌细胞，种于24孔板，体外培养48 h后，设置空白组、模型组、美托洛尔组及黄芪甲苷低、中、高剂量组。预先给予各组对心肌细胞有害的儿茶酚胺类物质异丙肾上腺素（ISO），造成体外原代心肌细胞损伤模型，空白组心肌细胞不加ISO；30 min后予各治疗组相应浓度的美托洛尔（2.336 μg·L-1）及黄芪甲苷低、中、高剂量（3，10，30 μmol·L-1）。BCA法检测总蛋白含量，蛋白质免疫印迹法（Western blot）检测原代心肌细胞的B细胞淋巴瘤/白血病-2（Bcl-2），Bcl-2相关蛋白X（Bax）及Bcl-2相关k蛋白（Bak）的表达，流式细胞学法检测各组细胞的凋亡率，Hochest/PI双染法镜下观察各组细胞细胞核的形态，从而检测细胞的凋亡情况。结果：与空白组比较，各给药组（10~40 μmol·L-1）原代心肌细胞的活力上调（P<0.05）；HOCHEST/PI双染结果显示，与模型组比较，美托洛尔组及黄芪甲苷中、高剂量亮蓝色凋亡细胞明显减少；细胞流式结果显示，与模型组比较，美托洛尔组凋亡率下调（P<0.01），黄芪甲苷中剂量组凋亡率下调（P<0.05），黄芪甲苷高剂量组凋亡率下调（P<0.01）；Bcl-2灰度值结果显示，与模型组比较，美托洛尔组Bcl-2的表达上调（P<0.01），黄芪甲苷低剂量组Bcl-2表达上调（P<0.05），黄芪甲苷中、高剂量组Bcl-2表达上调（P<0.01）；Bak灰度值结果显示，与模型组比较，美托洛尔组Bak的表达下调（P<0.05），黄芪甲苷高剂量组Bak表达下调（P<0.05）；Bax灰度值结果显示，与模型组比较，美托洛尔组Bax的表达下调（P<0.01），黄芪甲苷高剂量组Bax表达下调（P<0.05）。结论：黄芪甲苷对异丙肾上腺素诱导的大鼠原代心肌细胞损伤有抗凋亡作用。

Abstract

Objective: To study the anti-apoptosis effect of astragaloside Ⅳ on isoproterenol-induced rat primary myocardial cell injury. Method: Enzymatic lysis and differential velocity adherent technique were used to isolate primary myocardial cellsfrom postnatal heart of 1-3-day-old normal SD rats

inoculate them in 24 aperture planks and cultured for 48 h in vitro. Normal group

model group

metoprolol group and low

medium and high-dose astragaloside Ⅳ groups were set up. Isoprenaline (ISO) was given to cause heart failure in vitro

except for normal group. After 30 min

the corresponding concentrations of metoprolol (2.336 μg·L-1) and astragaloside Ⅳ (low dose 3 μmol·L-1

middle dose 10 μmol·L-1

high dose 30 μmol·L-1) were given to the treatment groups. The total protein content was determined with BCA method

the expressions of B cell lymphoma/lewkmia-2 (Bcl-2)

Bcl-2 associated X protein (Bax) and Bcl-2 associated K protein (Bak) in primary cardiomyocytes were detected by Western blot

theapoptosis rate in each group was detected by flow cytometry

and the cell nuclei in each group was observed by Hochest/PI double staining method. Result: Compared with the control group

the vitality of the primary cardiomyocytes in each administration group (10-40 μmol·L-1) was up-regulated (P<0.05). Hochest-PI double staining showed that compared with the model group

metoprolol group

and middle and high-dose astragaloside Ⅳ groupsshowed decreases in apoptoticbright blue cells.Flow cytometry showed that compared with the model group

the apoptosis rate of metoprolol group was down-regulated (P<0.01)

and the apoptosis rate of medium-dose astragaloside Ⅳ group was down-regulated (P<0.05)

and the apoptosis rate of high-dose astragaloside Ⅳ group was down-regulated (P<0.01). The gray value of Bcl-2 results showed that compared with model group

the expression of Bcl-2 in the metoprolol group was increased (P<0.01)

the expression of Bcl-2 was increased in astragaloside Ⅳ group (P<0.05)

and the expression of Bcl-2 was increased in high-dose astragaloside Ⅳ group (P<0.01). The gray value of Bak showed that compared with model group

the expression of Bak in the metoprolol group was down-regulated (P<0.05)

and the expression of Bak in high-dose astragaloside Ⅳ group was down-regulated (P<0.05). The gray value Bax showed that compared with model group

the expression of Bax in the metoprolol group was down-regulated (P<0.01)

and the expression of Bax in high-dose astragaloside Ⅳ group was down-regulated (P<0.05). Conclusion: Astragaloside Ⅳ has the anti-apoptosis effect on the injury of isoproterenol-induced rat primary cardiomyocytes.

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