肝细胞内相关mRNA抑制HBV复制与表达的功能验证

Functional verification of inhibiting HBV replication and expression by hepatocyte-related mRNA

目的验证肝细胞内抑制乙型肝炎病毒(HBV)复制与表达的相关mRNA。方法通过慢病毒介导的方式将过氧化物酶体增殖物激活受体α(PPARα)、核因子I/B(NFIB)及白细胞介素-8(IL-8)基因转入HepG2.2.15及HBV全基因组1.3倍体HepG2(HBV1.3P-HepG2)细胞模型中,以空白对照(NC)组为对照,转染48 h后采用qPCR检测mRNA表达及HBV DNA复制水平,化学发光法检测细胞上清液中HBsAg和HBeAg的表达,免疫荧光法检测细胞内HBsAg的表达。结果在HepG2.2.15和HBV1.3P-HepG2细胞模型中,PPARα能够促进HBV DNA复制和HBsAg表达,其HBV DNA的表达分别是NC组的1.46倍和1.27倍(P<0.05)。NFIB可以抑制HBV DNA复制和HBsAg表达,其HBV DNA的表达分别是NC组的0.76和0.55倍(P<0.05)。IL-8能够促进HBV DNA复制和HBsAg表达,其HBV DNA的表达分别是NC组的1.54倍和1.62倍(P<0.05)。结论肝细胞内PPARα和IL-8能够促进HBV复制与表达,NFIB可以抑制HBV复制与表达,可为后续研究提供实验室依据。

Objective To verify the mRNA that inhibits the replication and expression of hepatitis B virus(HBV) in hepatocytes. Methods The peroxisome proliferator activated receptor(PPAR) α(PPARα), nuclear factor B(NFIB) and interleukin-8(IL-8) genes were transferred into HepG2.2.15 and HBV whole genome 1.3 ploid HepG2(HBV1.3 P-HepG2) cell models by lentiviral mediation. 48 hours after transfection, mRNA expression and HBVDNA replication level were detected by qPCR with blank control group(NC) as control. The expression of HBsAg and HBeAg in the cell supernatant was detected by chemiluminescence, and the expression of HBsAg in the cells was detected by immunofluorescence. Results In HepG2.2.15 and HBV1.3 P-HepG2 cell models, PPARα could promote the replication of HBVDNA and the expression of HBsAg, and the expression of HBVDNA was 1.46 and 1.27 times higher than that of NC group(P<0.05). NFIB could inhibit the replication of HBVDNA and the expression of HBsAg, and the expression of HBVDNA was 0.76 and 0.55 times higher than that of NC group(P<0.05). IL-8 could promote the replication of HBVDNA and the expression of HBsAg, and the expression of HBVDNA was 1.54 times and 1.62 times higher than that of NC group(P<0.05). Conclusion PPARα and IL-8 in hepatocytes can promote the replication and expression of HBV, while NFIB can inhibit the replication and expression of HBV, which can provide laboratory basis for follow-up research.