摘要
目的:观察西罗莫司(又名雷帕霉素,Rap)对柯萨奇病毒B3(CVB3)感染的心肌成纤维细胞增殖及哺乳类雷帕霉素靶蛋白(mTOR)mRNA和蛋白表达的影响。方法:SD大鼠心肌成纤维细胞分对照组、CVB3组、Rap组及CVB3+Rap组,用重复感染率(MOI)为0.5噬斑形成单位(PFU)/cell的CVB3诱导心肌成纤维细胞,采用四氮唑蓝(MTT)法检测细胞增殖,RT-PCR及Western Blot分别检测其mTOR mRNA表达及mTOR蛋白表达。结果:MTT法测得对照组、CVB3组、Rap组及CVB3+Rap组的D值分别为0.55±s 0.09、0.66±0.09,0.42±0.09、0.36±0.12,CVB3组高于对照组,Rap组和CVB3+Rap组均低于对照组及CVB3组(P<0.05)。用RT-PCR检测上述4组的mTOR mRNA表达,mTOR/β-actin光密度比值分别为0.25±0.09、0.87±0.15、0.19±0.04、0.22±0.06,CVB3组高于对照组(P<0.05),Rap组和CVB3+Rap组低于对照组及CVB3组(P<0.05)。Western Blot显示mTOR蛋白表达结果与RT- PCR结果相符。结论:Rap可抑制CVB3病毒诱导的大鼠心肌成纤维细胞增殖及mTOR表达,Rap抑制细胞增殖可能是通过mTOR信号途径起作用的。
AIM： To observe the effects of sirolimus （Rap） on cell proliferation and the mammalian target of rapamycin （mTOR） expression in the rat myocardial fibroblasts infected by coxsackievirus B3 （CVB3）. METHODS： The myocardial fibroblasts cells were divided into control group, CVB3 infected group （CVB3 group）, Rap treated group （Rap group）, and the virus infected cells treated with Rap group （CVB3 ＋ Rap group）. CVB3 （multiplicity of infection （MOI） =0.5 plaque forming unit （PFU） /cell） was used to infect the primary cultured myocardial fibroblasts of SD rats. The cell proliferation was estimated by using MTT assay, and the mTOR expression was determined by RT-PCR and Western Blotting. RESULTS： The D values of the 4 groups were 0.55 ±s 0.09, 0.66 ± 0.09, 0.42 ± 0.09, and 0.36 ± 0.12 in control, CVB3, Rap, and CVB3 ＋ Rap groups, respectively. The D value of the CVB3 group was higher than that of control group and that in both Rap and CVB3 ＋ Rap group was lower than that in control or CVB3 group （P 〈 0.05）. The gray scale values of the mTOR in control, CVB3, Rap, and CVB3 ＋ Rap groups were 0.25 ± 0.09, 0.87 ± 0.15, 0.19 ± 0.04, 0.22 ± 0.06, respectively. The gray scale value of CVB3 group was higher than that of control group, and that in both Rap and CVB3 ＋ Rap group was lower than that in control or CVB3 group （P 〈 0.05）. The results of Western Blot were the same as that of RT-PCR （P 〈 0.05）. CONCLUSION： Rap can inhibit cell proliferation and mTOR expression in the rat myocardial fibroblasts infected by CVB3, and the mechanism may be associated with mTOR signal pathway.
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2007年第9期649-652,共4页
Chinese Journal of New Drugs and Clinical Remedies
基金
湖南省卫生厅科研基金(B2005-072)
2006年教育部留学回归启动基金资助
