大黄素体外抗人巨细胞病毒药效实验研究

Experiment effect research of emodin in vitro resistence human cytomegalovirus

  • 摘要: 目的 研究大黄素体外抗人巨细胞病毒(HCMV)的作用并探讨其抗病毒的分子机制。方法 采用噻唑蓝(MTT)法检测不同浓度的大黄素与更昔洛韦对人胚肺成纤维细胞存活率及其对HCMV抑制率的影响,采用曲线回归计算药物浓度与细胞存活率、病毒抑制率的回归方程,计算药物的半数毒性浓度(TC50)和病毒半数抑制浓度(IC50),以及治疗指数(TI),建立HCMV病毒感染细胞模型,予以更昔洛韦及大黄素干预,分为病毒对照组、大黄素组,采用实时荧光定量PCR方法检测各组细胞感染后0.5h﹑2h﹑24h即刻早期基因(ul122)﹑早期基因(ul54)与晚期基因(ul83) mRNA动态表达情况。结果 大黄素与更昔洛韦的TC50分别为75.96μg/ml和167.43μg/ml,IC50分别为7.31μg/ml和26.50μg/ml,TI分别为10.39和6.31;病毒感染后三个时间点检测大黄素组ul122mRNA表达量(0.61±0.06、1.62±0.16、2.16±0.18)与ul54mRNA表达量(0.82±0.04、4.23±0.08、970.33±52.04)均低于病毒对照组与更昔洛韦组;24h后大黄素组ul83mRNA表达量(1529.33±107.06)低于病毒对照组与更昔洛韦组。结论 大黄素的治疗指数高于更昔洛韦,大黄素具有良好的抗人巨细胞病毒HCMV的临床应用前景,同时大黄素在细胞水平上可抑制HCMV AD169株的即刻早期、早期、晚期基因的转录水平,表明大黄素对HCMV病毒基因的早期干预是其抗病毒的主要环节。

     

    Abstract: OBJECTIVE To study the antiviral effects of emodin on human cytomegalovirus and to explore the molecular mechanism through observation the effect of emodin on HCMV and viral gene transcription. METHODS MRC5 cell viability and HCMV inhibitory rate of emodin and ganciclovir in different dosage groups were detected with MTT method.ALL data were input to IBM SPSS STATISTICS 22.0 software for analyzing 50% toxic concentrations(TC50), 50% inhibition concentration(IC50) and therapeutic index(TI).HCMV AD169 weve established strain infected cells as the virus control group, emodin intervention infection cell model for emodin group and ganciclovir group,real-time fluorescent quantitative PCR method was wsed to detect virus immediate early genes ul122 mRNA, early genes ul54 mRNA, late genes ul83 mRNA dynamic expression of three groups after infection for 0.5h,2 hours and 24hours.RESULTS The TC50 of emodin and ganciclovir were 75.96μg/ml and 167.43 ug/ml; the IC50 were 7.31μg/ml and 26.50μg/ml;TI were 10.39 and 6.31μg/ml. 2 In different time of virus infection, ul122 and ul54 mRNA expression of emodin group were lower than virus control group and ganciclovir group. 24h after virus infection, ul83 mRNA expression of emodin group were lower than virus control group and ganciclovir group.CONCLUSION Therapeutic index of emodin is superior to ganciclovir, therefore emodin possesses HT5"SS a better application prospect on inhibiting human cytomegalovirus. In cellular level emodin can significantly inhibit HCMV AD169 strain of immediate early, early gene, late gene transcription level, show that emodin early intervention of HCMV virus gene is the main link of antiviral.

     

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