姜黄素抗人巨细胞病毒感染的体外实验研究

Activity of curcumin against human cytomegalovirus infection in vitro

本文旨在探讨姜黄素(Cur)体外抗人巨细胞病毒(HCMV)的作用。通过体外培养人胚肺成纤维细胞,采用四唑盐(MTS)方法检测Cur对细胞活力的影响。本研究分为对照组、HCMV组、HCMV+膦甲酸钠(PFA)组和HCMV+Cur组,通过蚀斑实验观察各组细胞病变效应(CPE),采用定量聚合酶链式反应(qPCR)检测各组HCMV的DNA拷贝数,蛋白免疫印迹检测HCMV不同时序蛋白的表达情况。结果发现,在细胞毒性实验中,与对照孔相比,当Cur浓度小于等于15μmol/L时,Cur对细胞生长和活力没有明显影响(P>0.05)。HCMV感染细胞5 d后,细胞开始出现CPE,随着时间延长,蚀斑数量增加。Cur可以减少CPE的产生,且呈剂量依赖性。HCMV感染细胞后,其DNA拷贝数和蛋白表达量逐渐增多,呈时间依赖性;Cur能够显著抑制HCMV的DNA复制,并下调HCMV蛋白表达水平,呈浓度依赖性,差异具有统计学意义(P<0.05)。综上,Cur通过抑制HCMV的DNA的复制和下调HCMV不同时序蛋白表达水平从而发挥抗HCMV活性。本研究为抗HCMV感染药物的研发提供新的实验依据。

This study aimed to investigate the effect of curcumin(Cur)against human cytomegalovirus(HCMV)in vitro.Human embryonic lung fibroblasts were cultured in vitro.The tetrazolium salt(MTS)method was used to detect the effects of Cur on cell viability.The cells were divided into control group,HCMV group,HCMV+(PFA)group and HCMV+Cur group in this study.The cytopathic effect(CPE)of each group was observed by plaque test,then the copy number of HCMV DNA in each group was detected by quantitative polymerase chain reaction(qPCR),and the expression of HCMV proteins in different sequence was detected by Western blot.The results showed that when the concentration of Cur was not higher than 15μmol/L,there was no significant change in cell growth and viability in the Cur group compared with the control group(P>0.05).After the cells were infected by HCMV for 5 d,the cells began to show CPE,and the number of plaques increased with time.Pretreatment with Cur significantly reduced CPE in a dosedependent manner.After the cells were infected by HCMV,the DNA copy number and protein expression gradually increased in a time-dependent manner.Pretreatment with Cur significantly inhibited HCMV DNA copies and downregulate HCMV protein expression levels in a concentration-dependent manner,and the difference was statistically significant(P<0.05).In conclusion,Cur may exert anti-HCMV activity by inhibiting the replication of HCMV DNA and down-regulating the expression levels of different sequence proteins of HCMV.This study provides a new experimental basis for the development of anti-HCMV infectious drugs.