杨梅苷通过microRNA-122对丙型肝炎病毒复制的影响

Effect of Myricitrin on Replication of Hepatitis C Virus through MicroRNA-122

目的:探讨杨梅苷通过微小RNA-122(microRNA-122,miR-122)对丙型肝炎病毒(hepatitis C virus, HCV)复制的作用。方法:通过建立Huh7-luciferase-miR-122稳定转染细胞系筛选对miR-122表达有影响的化合物;MTT法检测杨梅苷对Huh7细胞增殖的影响;实时荧光定量PCR(RT-qPCR)检测Huh7细胞内miR-122 mRNA的表达。通过瞬时转染HCV质粒进入Huh7细胞,RT-qPCR检测杨梅苷及干扰素-α(IFN-α)对HCV复制的影响。结果:与DMSO对照组比较,杨梅苷5μmol/L^50μmol/L能够明显抑制miR-122的表达(P<0.05或P<0.01),结果呈浓度依赖性;杨梅苷对Huh7细胞增殖无明显影响;杨梅苷10、25、50μmol/L对miR-122 mRNA的表达有着明显的抑制作用(P<0.05或P<0.01);杨梅苷25μmol/L能够抑制HCV的复制并且能够促进IFN-α对HCV复制的抑制作用(P<0.01)。结论:杨梅苷通过抑制miR-122的表达来抑制HCV的复制,并且可以促进IFN-α对HCV复制的抑制作用。

Objective:To explore the effect of myricitrin on the replication of hepatitis C virus(HCV) via microRNA-122(miR-122). Methods: Huh7-luciferase-miR-122 stable transfected cell line was established to screen compounds which could influence miR-122 expression. MTT assay was used to detect the effect of myricitrin on Huh7 cell proliferation. RT-qPCR was used to detect miR-122 mRNA expression in Huh7 cells. By transiently transfecting HCV plasmid into Huh7 cells, RT-qPCR was used to detect the effects of myricitrin and interferon-α(IFN-α) on HCV replication. Results: Compared with DMSO control group, the expression of miR-122 was inhibited by 5 μmol/L^50 μmol/L myricitrin(P<0.05 orP<0.01), in dose dependant manner. Myricitrin had no significant effect on the proliferation of Huh7 cells. Myricitrin(10、25、50 μmol/L) showed a significant inhibition on the expression of miR-122 mRNA(P<0.05 orP<0.01). 25 μmol/L Myricitrin inhibited the replication of HCV and promoted the inhibitory effect of IFN-α on the replication of HCV(P<0.01).Conclusion: Myricitrin inhibits the replication of HCV by down regulating the expression of miR-122, and promotes the inhibitory effect of IFN-α on the replication of HCV.