灵猫方对HepG2.2.15和HepAD38细胞La蛋白表达的影响

Effect of Lingmao Formula on the La protein expression in HepG2.2.15 and HepAD38 cells

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 study the inhibitive effects of Lingnmo Formula on cccDNA, pgRNA, sRNA and La-mRNA in HepG2.2.15 and HepAD38 cells, and discuss its mechanism on anti-hepatitis B virus（HBV）. Methods The aqueous extract of Lingnmo Formula was prepared. HepG2.2.15 and HepAD38 cells were intervened by the aqueous extract of Lingmao Formula at concentration of 500 and 1 000 mg/L respectively,the blank control group was treated with normal saline. The supematant of culture medium was collected after 6 days and the levels of HBsAg and HBeAg were detected by ELISA. The cells were collected and the total RNA was extracted, the expression levels of cccDNA, pgRNA, sRNA and La mRNA were detected by RT-PCR. Results Compared with the blank cantrol group, the secretion levels of HBsAg and HBeAg in HepG2.2.15 and HepAD38 cells of the Lingnmo Formula group were significantly decreased, the expression levels of intracellular cccDNA, pgRNA, sRNA and La mRNA were significantly decreased （P 〈 0.01 ）. Conclusion Lingnmo Formula may promote the degradation of pgRNA, sRNA and cccDNA by inhibiting La protein expression in HepG2.2.15 and HepAD38 cells, and result in the inhibition of HBV replication.