脂联素对体外培养新生牛脂肪细胞ADPN、HSL mRNA表达的影响

摘要

This experiement on adipocyte of calf was adopt to make primary culture, select monolayer growth well adipocyte of calf, then add concentration of the additives was as ADPN: 0,2. 5,5,15,30,50,100 μg/mL, after 12 h cultivation, abstract total RNA and test the effect of ADPN on the fat cell of ADPN mRNA and HSL mRNA expression by FQ-PCR method. With increasing the addition of ADPN, expression level of ADPN mRNA decreased; the expression level of HSL mRNA in fat cell was highest when the concentration of ADPN was 15 μg/mL in cell culture media,then decreased. The results showed as follows: the ADPN mRNA expression was regulated by negative regulator itself in adipocyte culture in vitro, but the appropriate amount concentration of ADPN could stimulate adipocyte HSL mRNA expression.%取单层生长状态良好的犊牛脂肪细胞,分别添加0、2.5、5、15、30、50、100 μg/mL外源牛重组脂联素(adiponectin,ADPN),培养12 h后提取总RNA,采用荧光定量PCR方法检测外源脂联素对脂肪细胞ADPN mRNA和HSL mRNA表达水平的影响.结果显示,随着ADPN添加量的增多,ADPN mRNA相对表达量呈下降趋势;添加ADPN 15 μg/mL时,HSLmRNA的表达量达到峰值,之后呈下降趋势.结果表明体外培养的脂肪细胞中ADPN mRNA表达量受自身的负调节,而添加适量的ADPN可刺激脂肪细胞HSL mRNA的表达.