CAJ | 학술논문

目的：研究赖氨大黄酸在体外对乙型肝炎病毒（HBV）增殖及病毒抗原表达的影响及分子机制，为研究赖氨大黄酸抗 HBV 作用机制奠定初步的实验依据。方法将 HepG2．2．15细胞分为实验组（分别加入5、10、20、40、80μmol／L 赖氨大黄酸）及不加赖氨大黄酸的对照组，分别培养24、48 h 收获上清液，采用四唑氮化合物（M TS）法检测细胞的增殖活性，采用 ELISA 法检测 HBsAg 和 HBeAg 的表达水平，实时定量 PCR（RT‐PCT ）检测其中 HBV DNA 的拷贝数及 miR‐210、miR‐185、miR‐199a‐3p 、miR‐370、miR‐196a 、miR‐184、miR‐217的表达水平。miR‐210、miR‐185、miR‐199a‐3p 、miR‐370、miR‐196a 、miR‐184、miR‐217反义链（ASO）分别转染 HepG2．2．15细胞后，M TS 法检测细胞的增殖活性，ELISA 法检测 HBsAg 和 HBeAg 的表达水平。结果在没有对 HepG2．2．15细胞的增殖活性产生影响的情况下，赖氨大黄酸抑制了 HBsAg 的产生和 HBV 的增殖。20μmol／L 的赖氨大黄酸明显促进了 miR‐210、miR‐185、miR‐199a‐3p 的表达，而抑制了 miR‐370的表达，与对照组相比较，差异具有统计学意义（P＜0．05）。结论赖氨大黄酸可能通过影响细胞内 miRNA 的表达抑制 HepG2．2．15细胞中 HBV 的增殖和HBsAg 的产生。
목적：연구뢰안대황산재체외대을형간염병독（HBV）증식급병독항원표체적영향급분자궤제，위연구뢰안대황산항 HBV 작용궤제전정초보적실험의거。방법장 HepG2．2．15세포분위실험조（분별가입5、10、20、40、80μmol／L 뢰안대황산）급불가뢰안대황산적대조조，분별배양24、48 h 수획상청액，채용사서담화합물（M TS）법검측세포적증식활성，채용 ELISA 법검측 HBsAg 화 HBeAg 적표체수평，실시정량 PCR（RT‐PCT ）검측기중 HBV DNA 적고패수급 miR‐210、miR‐185、miR‐199a‐3p 、miR‐370、miR‐196a 、miR‐184、miR‐217적표체수평。miR‐210、miR‐185、miR‐199a‐3p 、miR‐370、miR‐196a 、miR‐184、miR‐217반의련（ASO）분별전염 HepG2．2．15세포후，M TS 법검측세포적증식활성，ELISA 법검측 HBsAg 화 HBeAg 적표체수평。결과재몰유대 HepG2．2．15세포적증식활성산생영향적정황하，뢰안대황산억제료 HBsAg 적산생화 HBV 적증식。20μmol／L 적뢰안대황산명현촉진료 miR‐210、miR‐185、miR‐199a‐3p 적표체，이억제료 miR‐370적표체，여대조조상비교，차이구유통계학의의（P＜0．05）。결론뢰안대황산가능통과영향세포내 miRNA 적표체억제 HepG2．2．15세포중 HBV 적증식화HBsAg 적산생。
Objective To study the effect and molecular mechanisms of rhein lysinate(RHL) on the prolifera‐tion of hepatitis B virus(HBV) and viral antigen expression to establish the preliminary experimental evidence for re‐searching the anti‐HBV mechanism of RHL .Methods HepG2 .2 .15 cells were divided into the experimental groups (adding 5 ,10 ,20 ,40 ,80 μmol/L RHL) and the control group without adding RHL .After culturing for 24 ,48 h ,the supernatant was obtained .The cell proliferation activity was detected by the non‐radioactive cell proliferation assay (M TS) ,the expression level of HBsAg and HBeAg were detected by adopting the ELISA method ,the HBV DNA copy number and the expression levels of miR‐210 ,miR‐185 ,miR‐199a‐3p ,miR‐370 ,miR‐196a ,miR‐184 and miR‐217 were detected by real time quantitative PCR(RT PCT ) .HepG2 .2 .15 was transfected by miR‐210 ,miR‐185 ,miR‐199a‐3p ,miR‐370 ,miR‐196a ,miR‐184 and miR‐217ASO ,the cell proliferation activity was detected by the M TS method ,the expression levels of HBsAg and HBeAg were detected by rthe enzyme linked immunosorbent assay (ELISA) ,respectively .Results Without generating the effect on the cell proliferation activity ,RHL inhibited the generation of HBsAg and the HBV proliferation .The expression level of miR‐210 ,miR‐185 and miR‐199a‐3p was in‐creased by RHL(20 μmol/L) and the expression level of miR‐370 was inhibited by RHL(20 μmol/L) ,the differences compared with the control group were statistically significant(P < 0 .05) .Conclusion RHL is possible to suppress HBV proliferation and HBsAg generation by affecting the miRNA expression in HepG2 .2 .15 cells .