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目的:观察六月青皂苷(terpenoids of Liuyueqing,TLYQ)体外抗乙型肝炎病毒(HBV)共价闭合环状脱氧核糖核酸(cccDNA)复制的作用。方法:取HepG 2.2.15细胞,分为正常对照组、阿昔洛韦(ACV)阳性对照组、TLYQ低、中、高剂量组,各组细胞置CO2孵箱(37℃,5%CO2)中培养24 h后,TLYQ低、中、高剂量组加入TLYQ(终浓度分别为21.25,42.5,85 mg·L-1),阳性对照组加ACV(终浓度为1.02 mg·L-1),正常对照组加等体积培养液,以上每个浓度设3瓶,于培养72 h,144 h分别取细胞上清液,用实时荧光定量PCR法检测HBV ccDNA拷贝数。结果:在细胞培养72 h时,TLYQ高剂量组HBV cccDNA拷贝数即明显下降(P<0.05),在144 h高、中剂量组下降显著(P<0.05或P<0.01),低剂量组下降不明显。TLYQ作用呈明显的量效和时效反应关系。结论:TLYQ在体外有明显的抑制乙肝病毒的作用,可能是六月青主要活性成分之一。
Objective: To observe the effect of terpenoids of Liuyueqing (TLYQ) on the replication of hepatitis C virus (HBV) covalently closed circular deoxyribonucleic acid (cccDNA). Methods: HepG 2.2.15 cells were divided into normal control group, acyclovir (ACV) positive control group, TLYQ low, medium and high dose groups. Cells in each group were incubated in CO2 incubator (37 ℃, 5% CO2) Twenty-four hours after culture, TLYQ was added to TLYQ (final concentrations were 21.25, 42.5 and 85 mg · L-1, respectively), and positive control group was given ACV (final concentration was 1.02 mg · L-1) The normal control group plus equal volume of culture medium, each set above three bottles, cultured 72 h, 144 h were taken cell supernatant, real-time fluorescent quantitative PCR method to detect ccpDNA copy number. Results: At 72 h, the copy number of HBV cccDNA in TLYQ high dose group decreased significantly (P <0.05), decreased significantly in high and middle dose group at 144 h (P <0.05 or P <0.01), and decreased in low dose group Not obvious. The effect of TLYQ showed significant dose-response and time-dependent response. Conclusion: TLYQ can obviously inhibit the effect of hepatitis B virus in vitro and may be one of the main active components of Junqing green.