【摘要】 本研究旨在研究重组脂联素(rAdp)对皖南花猪肌内前体脂肪细胞脂联素(Adp)及其受体(AdipoR)、AMPK和PPARαmRNA表达的影响。选择5d皖南花猪背最长肌组织分离肌内前体脂肪细胞,增殖培养至80%融合,细胞分化后,用0,2和10mg/L rAdp分别处理12h和48h。油红O染色法鉴定脂肪细胞,MTT方法检测细胞活力;荧光定量RT-PCR方法检测脂肪细胞Adp、AdipoR1、AdipoR2、PPARα和AMPK mRNA表达量。结果显示,rAdp处理后,脂肪细胞活力总体有降低趋势,2mg/L处理24h达到显著水平(P<0.05);rAdp处理12h后,脂肪细胞AdipoR1和AdipoR2mRNA表达显著升高(P<0.05),AMPK mRNA表达量显著下降(P<0.05),但均无剂量依赖性。rAdp处理细胞48h,Adp mRNA表达显著下调(P<0.05),PPARαmRNA表达量显著升高(P<0.05),但剂量效应不明显;AMPK mRNA表达在10mg/L rAdp处理组显著升高(P<0.05)。此外,Compound C阻断rAdp处理引起的AMPK mRNA表达量下降。结果提示:重组脂联素处理猪肌内脂肪细胞有降低细胞活力作用,能上调AdipoR1、AdipoR2基因表达,影响PPARα和AMPK基因的表达,从而对肌内脂肪细胞起调节作用。更多还原

【Abstract】 This experiment was conducted to study the effect of recombinate adiponectin(rAdp)on adiponectin and its receptors,PPARαand AMPK mRNA expression in intramuscular preadipocytes of Wannanhua pig.Primary preadipocytes were separated from musculus longissimus tissue of Wannanhua pig at 5 days of age.Cells were cultured to 80% confluence.Adipocytes were treated with 0,2 and 10mg/L recombinate adiponectin(rAdp)for 12h or 48h,respectively.Oil-red O and MTT methods were used to identify adipocytes and to detect viability of cells.Adp,AdipoR1,AdipoR2,PPARαand AMPK mRNA levels were determined by real-time fluorescence quantitative RT-PCR.The results showed that the viability of cells had a reduced tendency,and decreased significantly by 2mg/L rAdp treatment for 24h(P<0.05).At 12hafter rAdp treatment,AdipoR1 and AdipoR2 mRNA expression of adipocytes were significantly increased(P <0.05),AMPK mRNA expression of adipocytes were significantly decreased(P<0.05),but all with no dose-dependent.At 48h after rAdp treatment,Adp mRNA expression of adipocytes were significantly decreased(P<0.05),PPARαmRNA expression were significantly decreased(P<0.05),but with no dose dependent;AMPK mRNA expression of adipocytes were significantly increased by 10mg/L rAdp treatment(P<0.05).In addition,the effects of rAdp treatment on AMPK mRNA expression were blocked by Compound C.The results indicated that recombinate adiponectin may depress the viability of intramuscular adipocytes.The effects of recombinate adiponectin(rAdp)on cultured intramuscular adipocytes mediated by up-regulating the mRNA expression of AdipoR1,AdipoR2 and affecting mRNA expression of the mRNA expression of PPARαand AMPK.更多还原