抗柯胶囊总皂苷对柯萨奇病毒B型诱导细胞凋亡的抑制作用

doi:10.19803/j.1672-8629.2022.04.09

中国药物警戒 ›› 2022, Vol. 19 ›› Issue (4): 384-389.
DOI: 10.19803/j.1672-8629.2022.04.09

抗柯胶囊总皂苷对柯萨奇病毒B型诱导细胞凋亡的抑制作用

张之斌¹, 姚荣妹^2,3,*, 马会霞², 包巨太², 郭姗姗⁴, 崔晓兰⁴

¹华北理工大学附属医院急诊科,河北唐山 063000;
²华北理工大学中医学院,河北唐山 063210;
³天津中医药大学中医药研究院,天津 301617;
⁴中国中医科学院中药研究所,北京 100700

收稿日期:2021-12-24 出版日期:2022-04-15 发布日期:2022-04-15
通讯作者: ^*姚荣妹,女,博士,讲师,中药药理学。E-mail：471268167@qq.com
作者简介:张之斌,男,硕士,主治医师,急诊感染性疾病治疗及药理学。
基金资助:
重大新药创制国家科技重大专项2009年度(2009ZX09103-442); 河北省中医药管理局中医药类科研计划项目（2016072）

Inhibitory effect of total saponins from Kangke capsules on apoptosis induced by Coxsackie virus B

ZHANG Zhibin¹, YAO Rongmei^2,3,*, MA Huixia², BAO Jutai², GUO Shanshan⁴, CUI Xiaolan⁴

¹Hospital Affiliated to North China University of Sciences and Technology, Tangshan Hebei 063000, China;
²College of Traditional Chinese Medicine, North China University of Sciences and Technology, Tangshan Hebei 063210, China;
³Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China;
⁴Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China

Received:2021-12-24 Online:2022-04-15 Published:2022-04-15

摘要/Abstract

摘要： 目的观察抗柯胶囊总皂苷抑制柯萨奇病毒B型（CVB）诱导细胞凋亡的作用,探讨其可能的机制。方法通过接种病毒液的方式建立体外细胞感染模型,采用细胞病变效应法（CPE）测定抗柯胶囊总皂苷对CVB₃、CVB₄、CVB₅株50%抑制浓度（IC₅₀）及治疗指数（TI）;CCK-8试剂盒检测细胞存活情况;TdT介导的dUTP缺口末端标记技术（TUNEL法）测定心肌细胞凋亡率;蛋白免疫印迹法（Western-blot）检测大鼠原代心肌细胞中Bcl-2、Bax蛋白表达情况。结果抗柯胶囊总皂苷可明显抑制CVB₃、CVB₄、CVB₅致细胞病变,对CVB₃、CVB₄、CVB₅的IC₅₀分别为223.34、315.48、306.53 μg·mL^-1,治疗指数分别为4.68、3.24、3.24;390.63、195.32、97.66 μg·mL^-1剂量组可明显提高3种病毒感染后心肌细胞的存活率,抑制心肌细胞凋亡。抗柯胶囊总皂苷对CVB₃病毒复制有明显抑制作用,390.63、195.32 μg·mL^-1 2个剂量组可使凋亡关键因子Bcl-2表达增加,Bax表达降低,Bcl-2/Bax比值升高,与病毒对照组比较有显著差异（P<0.05, P<0.01）。结论抗柯胶囊总皂苷可以改善CVB致心肌细胞病变,提高细胞存活率,减少细胞凋亡,其机制与抑制病毒复制、调控心肌细胞中凋亡相关蛋白表达相关。

关键词: 抗柯胶囊总皂苷, 柯萨奇病毒B型, 心肌细胞, 细胞凋亡

Abstract: Objective To observe the inhibitory effects of saponins from Kangke capsules on apoptosis caused by Coxsackievirus B (CVB), and to explore the mechanism. Methods CVB was inoculated into cardiomyocytes to establish an infection model in vitro. The cytopathic effect method (CPE) was adopted to determine the 50% inhibitory concentration (IC₅₀) and therapeutic index (TI) of saponins from Kangke capsules on CVB₃, CVB₄ and CVB₅. CCK-8 kit was used to detect cell viability. TdT-mediated dUTP notch end labeling technique (TUNEL) was used to determine the apoptosis rate of cardiomyocytes. Western-blot was used to study the expressions of Bcl-2 and Bax protein in primary rat cardiomyocytes. Results Saponins from Kangke capsules could significantly alleviate cytopathy induced by CVB. The IC₅₀ for CVB₃, CVB₄, and CVB₅ was 223.34, 315.48 and 306.53 μg·mL^-1 respectively, while the therapeutic index was 4.68, 3.24 and 3.24 respectively. The doses of 390.63, 195.32, and 97.66 μg·mL^-1 could increase the survival rate of cardiomyocytes after viral infection. Saponins from Kangke capsules could significantly inhibit viral replication and cell apoptosis after infection while increasing the expression of Bcl-2, decreasing the expression of Bax, and elevating the Bcl-2/Bax ratio. Conclusion Saponins from Kangke capsules can improve cytopathy caused by CVB infection, improve cell survival rate, and inhibit apoptosis. The mechanism is related to the regulation of apoptosis-related protein expressions in cardiomyocytes.

Key words: saponins from Kangke capsules, Coxsackievirus B, cardiomyocytes, apoptosis

中图分类号:

R978.7

张之斌, 姚荣妹, 马会霞, 包巨太, 郭姗姗, 崔晓兰. 抗柯胶囊总皂苷对柯萨奇病毒B型诱导细胞凋亡的抑制作用[J]. 中国药物警戒, 2022, 19(4): 384-389.

ZHANG Zhibin, YAO Rongmei, MA Huixia, BAO Jutai, GUO Shanshan, CUI Xiaolan. Inhibitory effect of total saponins from Kangke capsules on apoptosis induced by Coxsackie virus B[J]. Chinese Journal of Pharmacovigilance, 2022, 19(4): 384-389.

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抗柯胶囊总皂苷对柯萨奇病毒B型诱导细胞凋亡的抑制作用

Inhibitory effect of total saponins from Kangke capsules on apoptosis induced by Coxsackie virus B

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