滨蒿提取物抗乙肝病毒作用和急性毒性研究

Study on Anti-HBV Effect and Acute Toxicity of Artemisia Scoparia Extracts

目的:通过考察滨蒿提取物(BH)在载体细胞中对人乙肝病毒(HBV)DNA复制的干预作用,并观察其在小鼠口服给药的急性毒性反应,为滨蒿作为传统中药材治疗乙型肝炎的临床应用提供实验依据。方法:采用HBV转染人肝癌细胞的HepG2.2.15细胞模型,荧光定量PCR检测BH对细胞培养上清中HBV DNA表达的影响;酶联免疫法(ELISA)检测BH对细胞培养上清中乙肝病毒表面抗原(HBsAg)和e抗原(HBeAg)分泌的影响。以荧光定量PCR法检测乙肝病毒载体细胞HepG2.2.15细胞和HepAD38细胞内HBV DNA的表达量,观察BH对两种载体细胞中HBV复制的抑制作用。测定BH在小鼠灌胃给药的一日内最大给药量(MTD),观察给药后14d内出现的急性毒性反应。结果:BH对2.2.15细胞培养上清中HBV DNA表达有明显的抑制作用,IC_(50)平均为142.5μg/mL;其对2.2.15细胞上清中HBsAg和HBeAg的分泌也有明显的的抑制作用,IC_(50)平均为205.8μg/mL和105.3μg/mL;BH可明显抑制HepG2.2.15细胞内HBV DNA的复制,其IC_(50)为95.3μg/mL,还可明显抑制HepAD38细胞HBV DNA的高表达,其IC_(50)为97.1μg/mL。BH在小鼠一日内口服给药的MTD为12 g/kg·d(相当于原药材480g/kg·d),在14 d观察期内未出现明显的急性毒性反应。结论:滨蒿提取物在无毒浓度下对HepG2.2.15细胞内HBV DNA的稳定表达以及HepAD38细胞内HBV DNA的高表达均有良好的抗乙肝病毒作用,并且在小鼠口服给药安全性好。滨蒿提取物在抗乙肝病毒方面有较高的研究和应用价值。

Objective Observing the inhibitory effect of Artemisia scoparia extracts(BH)on HBV DNA replication in hepatitis B virus vector cells,and its security of oral administration,to provide experimental basis for the application of Artemisia artemisia as traditional Chinese medicine in the treatment of hepatitis B.Methods The effect of BH on the expression of HBV DNA in cell culture supernatant was detected by fluorescence quantitative PCR in HepG2.2.15 cell model transfected with HBV.Enzyme-linked immunosorbent assay(ELISA)was used to detect the secretion of HBsAg and HBeAg in cell supernatant.The expression of HBV DNA both in HepG2.2.15 cells and HepAD38 cells were detected by fluorescence quantitative PCR,and the inhibitory effect of BH on HBV replication in two kinds of HBV carrier cells was observed.The maximum dose(MTD)of BH by intragastric administration in a day was determined to observe the acute toxic reactions in mice.Results BH had obvious inhibitory effect on HBV DNA expression in supernatant of 2.2.15 cells,with an average IC_(50)of 142.5μg/mL.BH also significantly inhibited the secretion of HBsAg and HBeAg in the supernatant of 2.2.15 cells,with IC_(50)of 205.8μg/mL and 105.3μg/mL on average.BH can inhibit HBV DNA replication in HepG2.2.15 cells with IC_(50)of 95.3μg/mL,and also inhibit HBV DNA high expression in HepAD38 cells with IC_(50)of 97.1μg/mL.The oral MTD of BH in mice is 12 g/kg·d(equivalent to medicinal herb 480 g/kg·d),and no obvious acute toxic eaction was observed during the 14-day observation period.Conclusion Under none cytotoxic concentration,BH can inhibited markedly on both the stable expression of HBV DNA in HepG2.2.15 cells and the high expression of HBV DNA in HepAD38 cells,it has good anti-hepatitis B virus effect,and good security by oral administration.BH has high research and apply value in anti-hepatitis B virus.