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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。
HepG2.2.15 cells transfected with HBV gene were used as an in vitro model to study the effect of echinacoside on hepatitis B virus (HBV) replication and antigen expression. ELISA was used to determine the expression of HBsAg and HBeAg ) Levels; using quantitative PCR to detect HBV DNA levels. The effects of echinacoside on HBV replication were studied in vivo by using HBV transgenic mice. The levels of HBV DNA in the liver of transgenic mice were detected by fluorescence quantitative PCR. The levels of serum HBsAg and HBeAg were detected by ELISA. At the same time, the level of serum aminotransferase and the pathological changes of liver tissue were detected. Then the HBV transgenic mice were randomly divided into 5 groups: control group, lamivudine (50 mg · kg -1) group and echinacoside high and medium , Low dose (50,25,12.5 mg · kg-1) group. Gavage administered 1 day, continuous administration of 30 d. On the 31st day, the serum of mice was separated to detect the levels of HBV DNA, HBsAg and HBeAg. The DNA of liver tissue was extracted to determine the level of HBV DNA and the pathological examination of liver tissue. The results showed that echinacoside can significantly inhibit the expression of HBsAg and HBeAg in Hep G2.2.15 cells exposed to 50,100 mg · L-1 for 6 d, with the highest inhibition rates of 42.68% and 46.29%, respectively. At the same time, 100 mg · L -1 has an inhibitory effect on HBV DNA. In addition, echinacoside at a dosage of 50mg · kg-1 significantly inhibited HBsAg and HBeAg expression in HBV transgenic mice with the inhibitory rates of 42.82% and 29.12%, respectively, while significantly inhibiting the serum HBV DNA level in transgenic mice . It suggests that echinacoside has a strong inhibitory effect on HBV replication and antigen expression.