摘要
以HBV基因转染的HepG2.2.15细胞为体外模型,研究松果菊苷对乙肝病毒(HBV)复制和抗原表达的影响,采用ELISA法测定细胞上清乙肝表面抗原(HBsAg)和E抗原(HBeAg)水平;采用荧光定量PCR法检测HBV DNA水平。以HBV转基因小鼠为体内模型,研究松果菊苷对HBV复制的影响,首先采用荧光定量PCR法检测转基因小鼠肝组织HBV DNA水平,采用ELISA法检测小鼠血清HBs Ag和HBe Ag水平;同时检测血清转氨酶水平和肝组织病理变化;然后将HBV表达阳性的转基因小鼠随机分为5组:对照组、拉米夫定(50 mg·kg-1)组和松果菊苷高、中、低剂量(50,25,12.5 mg·kg-1)组。每天灌胃给药1次,连续给药30 d。第31天分离小鼠血清检测HBV DNA,HBsAg和HBeAg水平;提取肝组织DNA测定其HBV DNA水平,同时作肝组织病理检查。结果表明:松果菊苷在50,100 mg·L-1作用Hep G2.2.15细胞6 d后能显著抑制HBs Ag和HBeAg表达,其最高抑制率分别为42.68%,46.29%;同时在100 mg·L-1对HBV DNA具有抑制作用。此外,松果菊苷在50mg·kg-1剂量时能显著抑制HBV转基因小鼠HBsAg与HBeAg表达,其抑制率分别为42.82%,29.12%,同时对转基因小鼠血清HBV DNA水平有显著抑制作用。提示松果菊苷对HBV复制和抗原表达具有较强抑制作用。
To verify the effect of echinacoside on replication and antigen expression of hepatitis B virus（ HBV） by using HBVtransfected HepG2. 2. 15 cells as the in vitro model. The ELISA method was used to determine HBeAg and HBsAg levels in cellular supernatants. The effect of echinacoside on HBV replication was studied by using HBV transgenic mice as the in vivo model. First of all,the HBV DNA level in hepatic tissues was quantified with PCR method. Meanwhile,the serum transaminase levels and hepatic pathological changes were also evaluated. Subsequently,HBV transgenic mice were divided into five groups： the control group,the lamivudine group（ 50 mg·kg-1） and echinacoside high,medium and low dose group（ 50,25 and 12. 5 mg·kg-1）. The mice were orally administered with drugs once per day for 30 days. At the 31 stday,the mice serum was separated to measure HBsAg,HBeAg and HBV DNA. Additionally,the liver HBV DNA level and histopathological change were detected. The results indicated that echinacoside at 50 and 100 mg · L-1suppressed significantly HBsAg and HBeAg expressions on the sixth day,with the maximum inhibition ratios of42. 68% and 46. 29%; And echinacoside at 100 mg·L-1also showed an inhibitory effect on HBV DNA. Besides,echinacoside at 50mg·kg-1inhibited significantly HBs Ag and HBe Ag expressions of HBV transgenic mice,with the inhibition ratios of 42. 82% and29. 12%,and reduced markedly the serum HBV DNA level in HBV transgenic mice. In conclusion,the study suggested that echinacoside has a strong effect against HBV replication and antigen expression.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2015年第15期3047-3052,共6页
China Journal of Chinese Materia Medica
基金
教育部科技重点项目(212073)
浙江省中青年学科带头人学术攀登项目(pd2013187)
