毛花苷丙体外抑制单纯疱疹病毒Ⅰ型及对病毒合成基因表达的影响  

In vitro inhibition of herpes simplex virus type Ⅰ by lanatoside C and its effect on gene expression of virus synthesis

作　　者：叶凌风 吴松斌 熊东林 李容珍 YE Ling-feng;WU Song-bin;XIONG Dong-lin;LI Rong-zhen(Union Shenzhen Hospital,Huazhong University of Science and Technology,Shenzhen Guangdong 518000)

机构地区：[1]华中科技大学协和深圳医院,广东深圳518000

出　　处：《中南药学》2022年第11期2559-2564,共6页Central South Pharmacy

基　　金：深圳市南山区技术研发和创意设计项目(No.NS2021020)。

摘　　要：目的 探讨毛花苷丙体外抗单纯疱疹病毒Ⅰ型(HSV-1)的作用。方法 采用CCK8法计算毛花苷丙的半数有毒浓度(TC_(50))。细胞病变效应(CPE)和病毒滴度评价毛花苷丙的抗HSV-1作用。实时荧光定量PCR(RT-qPCR)和蛋白免疫印迹法探究毛花苷丙对HSV-1基因组转录、蛋白质合成的作用。结果 0.1 μmol·L^(-1)(TC_(50)=0.32 μmol·L^(-1))毛花苷丙可显著抑制HSV-1诱导的CPE。病毒滴度随着毛花苷丙处理浓度的增加显著下降(P<0.05),与CPE趋势一致。RT-qPCR和Western blot结果显示,HSV-1即刻早期基因(IE)ICP0、早期基因(E)UL23、潜伏相关基因LAT、糖蛋白G编码基因US4和病毒蛋白ICP0的表达均能被毛花苷丙抑制(P<0.05),且抑制呈剂量依赖性。毛花苷丙预处理可以抑制HSV-1诱导的CPE,且抑制呈剂量依赖性。结论 毛花苷丙通过抑制HSV-1基因组转录和蛋白质合成抵抗HSV-1感染,可以作为抗HSV-1的新型候选药物。Objective To determine the anti-herpes simplex virus type 1 (HSV-1) activity of lanatoside C in vitro.Methods The median toxic concentration (TC_(50)) was assessed by CCK8 assay.The cytopathic effect (CPE) and viral titers were used to evaluate the anti-HSV-1 effect of lanatoside C.Real-time quantitative PCR and Western blot were used to determine the effect of lanatoside C on HSV-1 genome transcription and protein synthesis.Results Lanatoside C (0.1 μmol·L^(-1),TC_(50)=0.32 μmol·L^(-1)) significantly inhibited HSV-1-induced CPE.The virus titer decreased significantly with the increase of lanatoside C (P<0.05),which was consistent with the CPE degree.RT-qPCR and Western blot showed that the expression of HSV-1 immediate early genes ICP0,early genes UL23,latency-associated gene LAT,glycoprotein G-encoding gene US4 and viral protein ICP0 were inhibited by lanatoside C in a dose-dependent manner (P<0.05).Pre-addition of lanatoside C inhibited HSV-1-induced CPE in a dose-dependent manner.Conclusion Lanatoside C resists HSV-1 infection by inhibiting HSV-1 genome transcription and protein synthesis,and can be used as a new candidate drug against HSV-1.

关 键 词：毛花苷丙 单纯疱疹病毒Ⅰ型 抗病毒 病毒滴度 

分 类 号：R285[医药卫生—中药学]