双氢青蒿素对C2株蓝氏贾第鞭毛虫Centrin基因mRNA表达水平的影响  

Effects of dihydroartemisinin on the level of centrin mRNA expression in C2 Giardialamblia

作　　者：余源[1] 王洋[1] 陈阳[1] 赵丽娜[1] 贺宝玲[1] 张亚粉[1] 田喜凤[1] 

机构地区：[1]河北联合大学生命科学学院,河北唐山063000

出　　处：《中国病原生物学杂志》2014年第4期360-363,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.30970313);河北省青年科学基金项目(C2012401039);河北联合大学培育基金项目(GP201308)

摘　　要：目的分析双氢青蒿素(dihydroartemisinin,DHA)对体外C2株蓝氏贾第鞭毛虫(Giardia lamblia)中心体蛋白(Centrin)基因mRNA表达水平的影响,探讨其对蓝氏贾第鞭毛虫的抑制作用。方法采用改良型TYI-S-33培养基培养C2株蓝氏贾第鞭毛虫,培养基内分别加入浓度为100μg/ml、200μg/ml的双氢青蒿素,以不含药物组作为阴性对照,分别培养2、4、8、12h后,收集各组虫体并提取总RNA,以此为模板,反转录合成cDNA,采用实时荧光定量RTPCR检测药物作用后Centrin基因mRNA表达水平的相对变化量。结果实时荧光定量RT-PCR检测显示,经双氢青蒿素作用虫体后Centrin基因mRNA表达水平显著降低,培养2、4、8、12h后100μg/ml药物组Centrin基因mRNA相对表达量分别为0.46、0.43、0.35和0.27,200μg/ml药物组分别为0.42、0.38、0.29和0.25。结论双氢青蒿素对C2株蓝氏贾第鞭毛虫Centrin基因mRNA的表达具有明显的抑制作用,且抑制作用随着药物浓度的增高和作用时间的延长而增强,提示双氢青蒿素对蓝氏贾第鞭毛虫具有明显的损伤作用,可供蓝氏贾第鞭毛虫病防治参考。Objectives To analyze the effects of dihydroartemisinin（DHA）on the level of Centrin mRNA expression in C2 Giardia lamblia and to discuss the control of C2 G.lamblia. Methods G.lamblia was cultured 2h,4h,8h,and12hin modified TYI-S-33medium containing 100μg/ml or 200μg/ml of DHA.G.lamblia was similarly cultured but without DHA to serve as the negative control.G.lamblia was collected and its RNA was extracted.RNA was reversetranscribed and cDNA was synthesized to serve as a template.The quantity of Centrin mRNA expression was determined using real-time PCR. Results Real-time PCR indicated that significantly less Centrin mRNA was expressed with DHA than with the control.The relative quantity of Centrin mRNA expression was 0.46when G.lamblia was cultured for 2h in medium containing 100μg/ml of DHA,0.43when it was cultured for 4h,0.35when it was cultured for 8h,and0.27when it was cultured for 12h.The relative quantity of Centrin mRNA expression was 0.42when G.lamblia was cultured for 2hin medium containing 200μg/ml of DHA,0.38when it was cultured for 4h,0.29when it was cultured for 8h,and 0.25when it was cultured for 12h. Conclusion Dihydroartemisinin markedly inhibited the level of Centrin mRNA expression in C2 G.lamblia.Dihydroartemisinin acts effectively on C2 G.lamblia,providing valuable reference data for the control of C2 G.lamblia.

关 键 词：C2株蓝氏贾第鞭毛虫 实时荧光定量RT-PCR 双氢青蒿素 中心体蛋白 

分 类 号：R382.2[医药卫生—医学寄生虫学]