大蒜新素对人巨细胞病毒感染人胚肺成纤维细胞Caspase-3表达的影响  

Effect of Allitridin on the Expression of Caspase3 in Human Embryo Lung Fibroblast Cells Infected by Human Cytomegalovirus

作　　者：孙涛涛[1] 方峰[1] 甄宏[1] 舒赛男[1] 刘瑾[1] 周玉峰[1] 李革[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科学系,武汉430030

出　　处：《医药导报》2007年第5期493-496,共4页Herald of Medicine

摘　　要：目的观察大蒜新素对人巨细胞病毒(HCMV)感染的人胚肺成纤维细胞(HELs)Caspase-3表达的影响,从细胞凋亡角度探讨大蒜新素抗HCMV效应的部分作用机制。方法用HCMV AD169毒株感染HELs,建立高感染复数(MO I)和低MO I感染细胞模型;设药物高、中、低剂量分别为9,6和3μg.mL-1,采用蛋白印迹法检测病毒感染和(或)药物处理后72 h细胞Caspase-3的表达水平。结果正常细胞仅见Caspase-3 P32表达;HCMV感染细胞P32表达显著增加,还出现活性片段P17条带,且高MO I组P17表达明显高于低MO I组;大蒜新素处理低MO I感染细胞P17表达呈上升趋势,P17/P32比值也明显增加;大蒜新素处理高MO I感染细胞P17表达下调,P17/P32比值也显著下降。结论HCMV无论高或低MO I感染均可诱导细胞凋亡增多,且高MO I诱导作用更强。大蒜新素一方面使低MO I感染细胞凋亡增多,另一方面抑制高MO I感染细胞的凋亡。前者可能有利于细胞清除病毒,后者则可减少细胞损伤而发挥保护作用。大蒜新素对HCMV感染细胞凋亡的调控呈双向性。Objective To investigate the effect of allitridin on the expression of caspase-3 in human embryo lung fibroblast ceils （HELs） infected by human cytomegalovirus （HCMV）, in order to explore the partial mechanism of allitridin against HCMV from the view of apoptosis. Methods HELs were infected by HCMV AD169 strain with high multiply of infection （ MOI = 2.5 ） and low MOI （0.25）, respectively. According to the maximum tolerance concentration （ MTC ） of allitri.din to HELs （9.6μg· mL^-1 ）, 9, 6 and 3μg· mL^-1 were determined as the high, medium and low dosage of the drug. The level of caspase-3 protein in each group was analyzed by using western blot assay at 72 h after viral infection and/or drug treatment. Results The normal HELs only expressed caspase-3 P32. Whereas, the expression of caspase-3 P32 increased after HCMV infection. Moreover, active fragment of P17 of caspase-3 appeared, and the expression of P17 in high MOI group was obviously stronger than that in low MOI group. Compared with infected controls, the active fragment of P17 was up-regulated in low MOI HELs treated with allitridin, and the ratio of P17 versus P32 also significantly increased. On the contrary the active fragment of P17 was down-regulated in high MOI HELs treated with drug, and the ratio of P17 versus P32 decreased as well. Conclusion HCMV infection could induce the apeptosis of HELs, and this effect was more intense in high MOI group than that in low MOI. Allitridin could up-regulate the apeptosis of low MOI HELs, while down-regulate that of high MOI cells, supposing that the former was inclined to eliminate the virus from the infected cells, and the latter might protect the cells from injury caused by excess cellular apeptosis. This suggests the regulation of aUitridin on HCMV-infected cells was the dual effect.

关 键 词：大蒜新素 人巨细胞病毒 细胞凋亡 CASPASE-3 

分 类 号：R286[医药卫生—中药学] R965.0[医药卫生—中医学]