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目的观察瘦素对体外培养的大鼠脂肪细胞乙酰辅酶A羧化酶(ACC)及单腺苷酸激活蛋白激酶α1(AMPKα1)mRNA表达的影响。方法取3只Wistar雄性大鼠附睾周围脂肪组织,进行原代脂肪基质细胞分离培养并诱导其成脂分化;采用50 nmol/L大鼠重组瘦素处理分化后的脂肪细胞6 h;逆转录-聚合酶链反应(RT-PCR)测定对照组和瘦素处理组脂肪细胞中ACC、AMPKα1 mRNA表达。结果原代脂肪基质细胞可诱导分化为脂肪细胞,计数分化率约为60%;瘦素处理组大鼠脂肪细胞的ACC mRNA表达水平(0.39±0.29)低于对照组(0.75±0.50),差异有统计学意义(P<0.05);对照组和瘦素处理组AMPKα1 mRNA表达水平分别为(0.67±0.39),(0.51±0.34),差异无统计学意义(P>0.05)。结论大鼠重组瘦素可抑制体外培养脂肪细胞的脂肪酸合成,其机制可能与AMPKαl无关。
Objective To observe the effect of leptin on the mRNA expression of acetylcholinesterase (ACC) and monoacitric acid activated protein kinase α1 (AMPKα1) in rat adipocytes cultured in vitro. Methods Peripheral epididymal adipose tissue was obtained from 3 Wistar male rats, and primary adipose stromal cells were isolated and cultured to induce adipogenic differentiation. Differentiated adipocytes were treated with 50 nmol / L rat recombinant leptin for 6 h. Reverse transcription - Polymerase chain reaction (RT-PCR) was used to measure the expression of ACC and AMPKα1 mRNA in adipocytes of control group and leptin-treated group. RESULTS: Primary adipose stromal cells were differentiated into adipocytes and the rate of differentiation was about 60%. The level of ACC mRNA (0.39 ± 0.29) in adipocytes of leptin-treated group was significantly lower than that of the control group (0.75 ± 0.50) (P <0.05). The levels of AMPKα1 mRNA in control group and leptin group were (0.67 ± 0.39) and (0.51 ± 0.34) respectively, with no significant difference (P> 0.05). Conclusion Rat leptin inhibits the fatty acid synthesis of cultured adipocytes in vitro. The mechanism may be independent of AMPKαl.