健肝散体外对拉米夫定耐药株病毒表达抑制及增敏作用的研究  

Study on the inhibitory and sensitizing effect of Jiangan powder on the expression of LMV resistant strain virus in vitro

作　　者：李堃 葛飞 孙茂秋 严展鹏[2] 仲婕 季瑜 朱方石 LI Kun;GE Fei;SUN Maoqiu;YAN Zhanpeng;ZHONG Jie;JI Yu;ZHU Fangshi(Department of Spleen and Stomach Diseases,Haian Hospital of Traditional Chinese Medicine,Haian 226600,China)

机构地区：[1]海安市中医院脾胃病科,江苏海安226600 [2]江苏省中医药研究院中西医结合临床研究室,江苏南京210046

出　　处：《陕西中医》2023年第9期1188-1192,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：江苏省自然科学基金资助面上项目(BK20181235);江苏省卫生健康委医学研究面上项目(M2022024);江苏省南通市科技计划指导性项目(MSZ20192);江苏省南通市卫生健康委员会科研立项课题(QA2020048)。

摘　　要：目的:探讨健肝散(JGP)对HepG2.2.15耐药株对拉米夫定(LMV)耐药的影响。方法:利用药物浓度递增冲击的方法,建立HepG2.2.15/LMV耐药细胞株模型,CCK8法检测耐药株、敏感株IC_(50)值,观察健肝散(JGP)含药血清对耐药株IC_(50)值影响;设置HepG2.2.15耐药株+正常大鼠血清组及HepG2.2.15耐药株+JGP含药血清组,荧光定量PCR检测两组细胞HBV-pgRNA表达水平,提取RNA后ELISA检测两组细胞上清HBsAg和HBeAg的表达水平。结果:①与HepG2.2.15细胞耐药株+正常大鼠血清组相比,HepG2.2.15细胞耐药株+JGP含药血清的IC_(50)值显著降低;②与耐药细胞株+正常血清组相比,健肝散含药血清可抑制HepG2.2.15 LMV耐药细胞株HBV-pgRNA的表达,差异有统计学意义(P<0.01);③与HepG2.2.15/LMV耐药细胞株添加正常血清组相比,JGP含药血清可抑制HepG2.2.15/LMV耐药细胞株HBsAg和HBeAg的表达,差异具有统计学意义(P<0.01)。结论:JGP体外可以增加耐药株对LMV敏感性,有效降低耐药株HBV的表达,对LMV耐药具有一定缓解作用。Objective:To investigate the effect of Jiangan powder(JGP)on lamivudine(LMV)resistance in HepG2.2.15 resistant strains.Methods:The drug resistant cell line model of HepG2.2.15/LMV was established by the method of increasing drug concentration.The IC_(50)values of drug resistant and sensitive strains were detected by CCK8 method,and the effect of JGP containing serum on the IC_(50)value of drug resistant strains was observed.The HepG2.2.15 resistant strain+normal rat serum group and the HepG2.2.15 drug resistant strain+JGP drug containing serum group were set.The expression level of HBV-pgRNA in the cells of the two groups was detected by fluorescence quantitative PCR,and the expression levels of HBsAg and HBeAg in the supernatant of the two groups were detected by ELISA after RNA extraction.Results:①Compared with the drug-resistant HepG2.2.15 cell line+normal rat serum group,the IC_(50)value of the drug-containing serum of the drug-resistant HepG2.2.15 cell line+JGP decreased significantly.②Compared with the drug-resistant cell line+normal serum group,the drug-containing serum of Jiangan powder could inhibit the expression of HBV-pgRNA in HepG2.2.15 LMV drug-resistant cell line(P<0.01).③Compared with the group of HepG2.2.15/LMV drug-resistant cell line supplemented with normal serum,JGP could inhibit the expression of HBsAg and HBeAg of HepG2.2.15/LMV drug-resistant cell line,and there was significant difference.Conclusion:JGP can increase the sensitivity of drug-resistant strains to LMV in vitro,effectively reduce the expression of HBV in drug-resistant strains,and alleviate the drug resistance of hepatitis B lamivudine(LMV)to a certain extent.

关 键 词：乙肝 健肝散 拉米夫定 耐药 增敏 细胞实验 HEPG2.2.15 抗病毒作用 

分 类 号：R512.62[医药卫生—内科学]