摘要
目的:研究合欢饮的水提物抗HCV病毒活性及其机制。方法:采用Huh7.5.1细胞建立HCV2a病毒感染模型;通过CellTiter-Glo法测定吸光度值计算得到细胞存活率(%)和RenillaLuciferaseAssayKit法测定Rluc值算得到抑制率(%)、CC_(50)、EC_(50)并计算SI值;通过检测荧光素酶活性观察病毒72 h内对不同浓度合欢饮的耐药性;应用Western-blot检测NS5A、NS3及NS5B蛋白表达水平。结果:合欢饮水提物抗HCV2a的CC_(50)、EC_(50)及SI分别为132.50、1.90、67.90μg/mL。在24、48、72 h给药后的EC_(50)分别为18.0、5.8、2.3μg/mL。在给药浓度范围内,合欢饮水提物对NS5A及NS5B蛋白的表达均有抑制作用且呈一定的量效关系,但对NS3蛋白表达的作用不明显。结论:合欢饮的水提物可能通过改变NS5A及NS5B蛋白表达水平来抑制HCV2a病毒的复制,且长时间给药病毒无耐药性。
Objective:To study the anti-HCV activity of Hehuan Yin aqueous extracts and its mechanism.Methods:Huh7.5.1 cells were used to establish the HCV2a virus infection models.Celltiter-GLO Assayand Renilla Luciferase Assay Kits were used to determine the cell survival rate(%),CC_(50),EC_(50),respectively.SI value was also calculated.The drug resistance of the virus to different concentrations of Hehuan Yin within 72 h was also determined.Protein expression levels of NS5A,NS3 and NS5B were detected by Western-blotting.Results:the CC50,EC50 and SI of Hehuan Yin against HCV2a was 132.50μg/mL,1.90μg/mL and 67.90μg/mL,respectively.The EC50 after 24 h,48 h and 72 h administration was 18.0μg/mL,5.8μg/mL and 2.3μg/mL,respectively.Within the range of administrated drug concentrations,the aqueous extract of Hehuan Yin had inhibitory effects on the expression of NS5A and NS5B proteins in a dose-dependent trend,but it had no obvious effects on the expression of NS3 protein.Conclusion:The aqueous extract of Hehuan Yin may inhibit the replication of HCV2a virus by changing the protein expression levels of NS5A and NS5B,and the virus has no tolerance to the aqueous extract of Hehuan Yin.
作者
巫园园
梁泳淋
陈飞龙
汤庆发
WU Yuan-yuan;LIANG Yong-lin;CHEN Fei-long;TANG Qing-fa(School of Traditional Chinese Medicine,Southern Medical University,Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics,Guangdong Provincial Engineering Laboratory of Chinese Medicine Preparation Technology,Guangzhou 510515,China)
出处
《海南医学院学报》
CAS
2022年第1期1-5,共5页
Journal of Hainan Medical University
基金
广东省自然基金(基于谱效关联分析的合欢皮抑制丙型肝炎病毒活性的有效组分研究2018A030313308)。
