重组脂联素对皖南花猪脂肪细胞脂联素及其受体mRNA表达的影响

脂联素（Adp）是主要由脂肪组织分泌的细胞因子,有重要的生理作用。本试验旨在研究重组脂联素（rAdp）对皖南花猪脂肪细胞脂联素及其受体2,AMP激活蛋白激酶（AMPK）、过氧化物增殖剂活化受体α（PPARα）mR-NA表达的影响。选择10d皖南花猪皮下脂肪组织分离前体脂肪细胞,增殖培养至80%融合后换分化培养基培养,细胞分化后用0、2和10mg/L rAdp分别处理12和48h。油红O染色法鉴定脂肪细胞,MTT方法检测细胞活力;酶法测定培养液中甘油释放量,荧光定量RT-PCR方法检测脂肪细胞脂联素（Adp）、脂联素受体1（AdpR1）、脂联素受体2（AdpR2）、PPARα和AMPK mRNA表达。结果显示,rAdp处理后,脂肪细胞活力总体有降低趋势,10mg/L处理48h达到显著水平（P〈0.05）;rAdp处理对甘油释放的抑制作用未达到差异水平。rAdp处理12h后,脂肪细胞AdpR1和AdpR2mRNA表达显著升高（P〈0.01）,但无剂量依赖性;rAdp处理48h后,脂肪细胞AdpmRNA表达显著下降（P〈0.05）。rAdp处理12h后,脂肪细胞PPARαmRNA表达显著升高（P〈0.01）,且有剂量效应性;而AMP AMPK mRNA表达均无显著性变化。结果提示,重组脂联素处理猪原代脂肪细胞有降低细胞活力和抑制脂肪细胞甘油释放量的趋势,能显著上调AdpR1、AdpR2和PPARα基因的表达,从而刺激脂肪酸氧化和甘油三酯的水解作用。

Effect of recombinate adiponectin on adiponectin and its receptors mRNA expression in adipocyte of Wannanhua pigs in vitro

Adiponectin（Adp） is a cytokine secreted mainly by adipose tissue,which has important physiological functions.This experiment was conducted to study the effect of recombinate adiponectin（rAdp） on adiponectin and its receptors,peroxisome proliferator activated receptor α（PPARα） and AMP-activated protein kinase（AMPK） mRNA expression in adipocytes of Wannanhua pig.Primary adipocytes were separated from subcutaneous adipose tissue of Wannanhua pig at 10 days of age.Cells were cultured to 80% confluence,and then cultured by differentiation media.Adipocytes were treated with 0,2 and 10 mg/L rcombinate adiponectin（rAdp） respectively for 12 or 48 h.Oil-red O and MTT methods were used to identify adipocytes and to detect viability of cells.The glycerol contents in cultured media was also detected.Adp,AdpR1,AdpR2,PPARα and 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 10 mg/L rAdp treatment for 48 h（P〈0.05）.Less glycerol were released from cells in both experimental groups than that in control group,but no significant difference.After rAdp treatment for 12 h,AdpR1 and AdpR2 mRNA expression of adipocytes were significantly increased（P〈0.01）,but with no dose dependent.Adp mRNA expression of adipocytes were significantly decreased after rAdp treatment for 48 h（P〈0.05）.PPARα mRNA expression were significantly increased after rAdp treatment for 12 h,and with dose dependent.AMPK mRNA expression were no significantly change in all experimental groups after 12 or 48 h.The results indicate that recombinate adiponectin may depress the viability of adipocytes and inhibit glycerol releasing of adipocytes.The acute rAdp treatment could increase fatty acid oxidation and lipolytic activity of adipocytes by up-regulating mRNA expression of AdpR1,AdpR2,and PPARα in adipocytes.