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目的探讨3种不同外源性脂肪酸对大鼠原代肝细胞糖代谢相关的葡萄糖激酶(GCK)、固醇调节元件结合蛋白-1c(SREBP-1c)基因表达的影响,为饮食预防糖尿病提供参考依据。方法用改良二步胶原酶灌注法提取SD大鼠的肝脏细胞并进行培养;根据预实验的结果确定脂肪酸的用量;分别用0、3.2和12.8 g/ml的软脂酸(PA)、油酸(OA)和二十二碳六烯酸(DHA)处理细胞6 h;采取RT-PCR法检测GCK mRNA和SREBP-1c mRNA的表达量。结果①高剂量脂肪酸(12.8!g/ml)处理时,PA组和OA组GCK mRNA表达量较对照组分别升高了40.14%和36.58%,SREBP-1c mRNA表达量较对照组分别升高了48.59%和29.12%,差异均有统计学意义(P<0.05)。②低剂量脂肪酸(3.2!g/ml)处理时,PA组和DHA组SREBP-1c mRNA表达量较对照组分别降低了23.13%和17.64%,差异均有统计学意义(P<0.05)。结论外源性脂肪酸的种类和剂量都能够影响大鼠原代肝细胞中GCK和SREBP-1c基因的表达。
Objective To investigate the effect of three different exogenous fatty acids on gene expression of glucokinase (GCK) and steroid regulatory element-binding protein-1c (SREBP-1c) in primary hepatocytes of rats and provide a reference for the prevention of diabetes by diet in accordance with. Methods The hepatic cells of SD rats were extracted and cultured by modified two-step collagenase perfusion method. The amount of fatty acids was determined according to the results of the pre-experiment. Palmitic acid (PA) at 0, 3.2 and 12.8 g / ml, (OA) and docosahexaenoic acid (DHA) for 6 h. The expressions of GCK mRNA and SREBP-1c mRNA were detected by RT-PCR. Results ① At the high dose of fatty acid (12.8μg / ml), the expression of GCK mRNA in PA group and OA group increased by 40.14% and 36.58% respectively compared with the control group, and the expression of SREBP-1c mRNA increased 48.59% and 29.12% respectively, with statistical significance (P <0.05). ② The mRNA expression of SREBP-1c in PA group and DHA group decreased by 23.13% and 17.64% compared with the control group when treated with low dose fatty acid (3.2μg / ml), the difference was statistically significant (P <0.05). Conclusion Exogenous fatty acid types and doses can affect the expression of GCK and SREBP-1c genes in rat primary hepatocytes.