苦参碱类生物碱联合胸腺肽抗HBV作用研究  

Thymopolypeptides combined with matrine type alkaloids suppress HBV replication

作　　者：刘晓琼[1,2] 沈宏辉[2] 陈佳欣[2] 柏兆方[2] 王伽伯[2] 肖小河[2] 

机构地区：[1]江西中医药大学药学院,江西南昌330004 [2]解放军302医院全军中药研究所,北京100039

出　　处：《中国中药杂志》2016年第7期1275-1281,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(8133090);山东省协同创新项目(XTCX2014B01-06)

摘　　要：该文主要研究4种苦参碱类生物碱分别联合胸腺肽在Hep G2.2.15细胞上的抗病毒效果。分别将氧化苦参碱、苦参碱、槐果碱、槐定碱（浓度均为0.2 mmol·L-1）联合胸腺肽（0.025,0.1 g·L-1）于Hep G2.2.15细胞作用48,72 h后收集细胞及上清。采用CCK-8法测定各实验组细胞的（活性）来评价药物对Hep G2.2.15细胞的毒性作用;采用酶联免疫吸附试验（ELISA）测定实验组上清液中HBs Ag,HBe Ag的含量;采用荧光定量PCR法分别检测细胞上清和胞内HBV DNA水平;采用CBA方法检测上清液中细胞因子IFN-α的表达量。结果显示,单独胸腺肽0.025~0.4 g·L-1对细胞没有毒性,在此浓度范围内联合0.2 mmol·L-1苦参碱类生物碱对细胞没有毒性;苦参碱类生物碱联合胸腺肽抗HBV效果优于单独用碱或胸腺肽;单独0.025 g·L-1胸腺肽组对胞内HBV DNA的抑制效果优于单独0.1 g·L-1胸腺肽组,而对于上清HBe Ag的抑制效果则相反;0.1 g·L-1胸腺肽联合碱和0.025 g·L-1胸腺肽联合碱抗HBV效果相当,4个生物碱和胸腺肽联合抗HBV效果无统计学差异;总体来看,72 h抗HBV效果优于48 h（P〈0.05）;联合用药之后IFN-α的分泌量升高,和其他4个指标的变化成正相关（P〈0.05）。结果表明,氧化苦参碱、苦参碱、槐果碱、槐定碱联合胸腺肽可通过促进IFN-α的表达来抑制Hep G2.2.15细胞HBs Ag,HBe Ag的分泌和HBV DNA的复制,进而促进抗病毒作用。To investigate the antiviral effect of thymopolypeptides combined with 4 kinds of matrine type alkaloids on Hep G2. 2. 15 cells,oxymatrine,sophocarpidine,sophocarpine,and sophoridine（ at concentration of 0. 2 mmol·L- 1respectively） were respectively combined with thymopolypeptides（ 0. 025,0. 1 g·L- 1）,and after 48 h and 72 h treatment on Hep G2. 2. 15 cells,the cells and supernatants were collected. The cells activity in various groups was determined by CCK-8 method to evaluate the toxic effects of the drugs on Hep G2. 2. 15 cells. Enzyme linked immunosorbent assay（ ELISA） was used to determine HBe Ag and HBs Ag levels in cellular supernatants. HBV DNA levels in cellular supernatants andcells were quantified with fluorogenic quantitative PCR method; and the expression level of IFN-α in supernatants was detected with CBA method. The results indicated that single thymopolypeptides at 0. 025-0. 4 g·L- 1had no toxicity to cells. Thymopolypeptides in this concentration range combined with 0. 2 mmol·L- 1matrine type alkaloids also had no toxicity to cells. Anti-HBV activity of drug combination was better than that of alkali or thymopolypeptides alone. Thymopolypeptides at 0. 025 g·L- 1had better inhibitory effect than thymopolypeptides at 0. 1 g·L- 1on intracellular HBV DNA expression,but the inhibitory effect on supernatant HBe Ag level was on the contrary. Anti-HBV activity was similar between alkaloids combined with 0. 1 g·L- 1and alkaloids combined with 0. 025 g·L- 1. There was no statistical difference in anti-HBV effect between various combined groups（ P 0. 05）. In general,72 h anti-HBV effect was better than 48 h anti-HBV effect（ P 0. 05）. The expression of IFN-α was increased after drug combination,with positive correlation to the changes of other four indicators（ P 0. 05）. In conclusion,oxymatrine,sophocarpidine,sophocarpine and sophoridine combined with thymopolypeptides could inhibit HBs Ag and HBe Ag secretion in Hep G2. 2. 15 cells and HBV DNA replication,and further promo

关 键 词：胸腺肽 苦参碱类生物碱 HEPG2.2.15细胞株 乙肝病毒 

分 类 号：R285[医药卫生—中药学]