【摘要】 目的:探讨血红素氧化酶1(hemeoxygenase,HO1)基因转染对大鼠肝细胞系BRL的抗凋亡作用。方法:体外培养的BRL细胞转染HO1基因后(对照组采用转染空载体pcDNA3的BRL细胞),应用RTPCR检测肝细胞系BRL中HO1基因的表达。用肿瘤坏死因子α(TNFα)刺激细胞,采用流式细胞仪(FACS)进行细胞计数,TUNEL染色等方法检测细胞的凋亡情况。结果:转染的肝细胞系BRL中均有HO1mRNA表达。转染HO1的BRL细胞经TNFα诱导凋亡后,细胞凋亡率(55.42%±3.12%)明显低于同等刺激下转染pcDNA3的BRL细胞(81.29%±3.08%)(P<0.05)。结论:BRL肝细胞系中HO1表现出较强的抗凋亡能力,为以细胞凋亡为主要病理表现的肝脏缺血再灌注损伤的临床治疗提供了新思路。更多还原

【Abstract】 Aim: To investigate the antiapoptotic effect of HO-1 gene transfection in the cell line BRL derived from liver in vitro.Methods:HO-1 was transfected into the cell line BRL by liposome, and then the positive clone was selected by G418. RTGAAB2PCR and Western Blot were used to detect the expression of HO-1 protein. Then the cells overexpressing HO-1 were exposed to TNF-α stimulater, the mortality was measured by fluorescence-activated cell sorting (FACS), in addition,the TUNEL staining was also used to detect the degree of cell apoptosis. Results: The liver BRL cell line that had been transfected proved to be HO-1 positive. The apoptosis rate of BRL cells was 55.42%±3.12%, much lower than that of control group that transfected with pcDNA3 (81.29%±3.08%) under the stimuli of TNF-α(P<0.05). Conclusion: HO-1 gene can exhibit an anti-apoptosis effect in BRL cell line, and may play a key role in the treatment of diseases in liver ischemic/reperfusion in which apoptosis is their main manifestation.更多还原