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由于膳食原因,人体摄入ω-6PUFAs/ω-3PUFAs比例过高,脂类代谢严重失衡.鉴于ω-3PUFAs获取困难而且人体无催化ω-6PUFAs向ω-3PUFAs转化的ω-3多不饱和脂肪酸脱氢酶,本研究体外扩增来源于秀丽线虫(Caenorhabditis elegans)的ω-3多不饱和脂肪酸脱氢酶基因(fat-l)cDNA,构建了真核表达载体pEGFPC1-fat-1,将该基因转染入小鼠胚胎成纤维细胞;激发荧光与RT-PCR方法检测转染pEGFPC1-fat-1细胞表达该基因,气相色谱分析显示该转染细胞中ω-6PUFAs/ω-3PUFAs的比例降低;细胞抑制率实验显示转染细胞的MTT吸光值升高(P<0.05);双染法流式细胞仪分析转染细胞凋亡降低,结果表明fat-1基因即使在高浓度ω-6PUFAs的细胞毒作用下,依然能够发挥将ω-6PUFAs脱氢转化为与ω-3PUFAs的生理功能,抑制3T3细胞凋亡,促进细胞生长增殖,实现了较强的细胞保护功能.
Due to dietary reasons, the excess of omega-6 PUFAs / omega-3 PUFAs in the human body causes a serious imbalance of lipid metabolism. In view of the difficulty in obtaining omega-3 PUFAs and the lack of omega-3 polyunsaturated fatty acids in the human body that do not catalyze the conversion of omega-6 PUFAs to omega-3 PUFAs In this study, the omega-3 polyunsaturated fatty acid desaturase gene (fat-1) cDNA derived from Caenorhabditis elegans was amplified in vitro and the eukaryotic expression vector pEGFPC1-fat-1 was constructed. The gene was transfected into mouse embryonic fibroblasts. The expression of the gene was detected by fluorescence and RT-PCR in pEGFPC1-fat-1 cells. Gas chromatography analysis showed that the percentage of ω-6PUFAs / ω-3PUFAs in the transfected cells was decreased (P <0.05). The apoptosis of transfected cells was reduced by double staining and flow cytometry. The results showed that the fat-1 gene was expressed in high concentrations of ω-6PUFAs Cytotoxicity, still can play the physiological function of ω-6PUFAs dehydrogenation and ω-3PUFAs, inhibit 3T3 cell apoptosis, promote cell growth and proliferation, to achieve a strong cell protection.