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目的研究灵猫方对HepG2.2.15和HepAD38细胞cccDNA、pgRNA、sRNA和La-mRNA抑制作用,探讨灵猫方抗乙肝病毒(HBV)的作用机制。方法制备灵猫方水提物,500 mg/L和1 000 mg/L灵猫方水提物分别干预HepG2.2.15和HepAD38细胞,以0.9%Na Cl溶液作为空白对照组,6 d后收集培养液上清,ELISA法检测HBsAg和HBeAg水平;收集细胞提取总RNA,RT-PCR法检测细胞内cccDNA、pgRNA、sRNA和LamRNA表达水平。结果与空白对照组比较,灵猫方组的HepG2.2.15和HepAD38细胞的HBsAg和HBeAg分泌水平明显降低,细胞内cccDNA、pgRNA、sRNA和LamRNA表达水平明显降低(P<0.01)。结论灵猫方可能通过抑制HepG2.2.15和HepAD38细胞内La蛋白的表达而促进pgRNA、sRNA和cccDNA的降解,从而抑制HBV的复制。
Objective To investigate the inhibitory effect of Lingcaofang on the cccDNA, pgRNA, sRNA and La-mRNA in HepG2.2.15 and HepAD38 cells and to explore the mechanism of the effect of Lingcaofang on Hepatitis B virus (HBV). Methods The water extract of civet cats, 500 mg / L and 1 000 mg / L of Codonopsis extracts were used to interfere HepG2.2.15 and HepAD38 cells, 0.9% NaCl solution was used as blank control group, 6 days after the collection of culture medium The levels of HBsAg and HBeAg were detected by ELISA. The total RNA was collected and the expression of cccDNA, pgRNA, sRNA and LamRNA were detected by RT-PCR. Results Compared with the blank control group, the levels of HBsAg and HBeAg in HepG2.2.15 and HepAD38 cells of Lingmaifang group were significantly decreased, and the expressions of cccDNA, pgRNA, sRNA and LamRNA were significantly decreased (P <0.01). Conclusion Cincao prescription may inhibit the replication of HBV by inhibiting the expression of La protein in HepG2.2.15 and HepAD38 cells and promoting the degradation of pgRNA, sRNA and cccDNA.