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

作　　者：张之斌 姚荣妹 马会霞 包巨太 郭姗姗[4] 崔晓兰[4] ZHANG Zhibin;YAO Rongmei;MA Huixia;BAO Jutai;GUO Shanshan;CUI Xiaolan(Hospital Affiliated to North China University of Sciences and Technology,Tangshan Hebei 063000,China;College of Traditional Chinese Medicine,North China University of Sciences and Technology,Tangshan Hebei 063210,China;Institute of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[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

出　　处：《中国药物警戒》2022年第4期384-389,共6页Chinese Journal of Pharmacovigilance

基　　金：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)。

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CVB_(3)、CVB_(4)、CVB_(5)株50%抑制浓度(IC_(50))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CVB_(3)、CVB_(4)、CVB_(5)致细胞病变,对CVB_(3)、CVB_(4)、CVB_(5)的IC_(50)分别为223.34、315.48、306.53μg·mL^(-1),治疗指数分别为4.68、3.24、3.24;390.63、195.32、97.66μg·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CVB_(3)病毒复制有明显抑制作用,390.63、195.32μg·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。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(IC_(50))and therapeutic index(TI)of saponins from Kangke capsules on CVB_(3),CVB_(4) and CVB_(5).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 IC_(50) for CVB_(3),CVB_(4),and CVB_(5) was 223.34,315.48 and 306.53μg·mL^(-1) respectively,while the therapeutic index was 4.68,3.24 and 3.24 respectively.The doses of 390.63,195.32,and 97.66μg·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.

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 

分 类 号：R978.7[医药卫生—药品]