试验首先建立了30日龄猪皮下脂肪和背最长肌组织块体外培养体系,共设2个处理,处理1为阴性对照(添加0.1mmol/L牛血清白蛋白,BSA),处理2添加100μmol/L的t10,c12-CLA,每个处理设6个平行,接种时即加入BSA与t10,c12-CLA进行处理至第10天,试验结束后收集细胞保存备用。采用荧光定量PCR技术及试剂盒检测研究t10,c12-CLA对猪皮下和背最长肌脂肪代谢关键酶(FAS、ME、LPL、HSL)和激素(INSR、GHR)及甘油三酯(TG)合成的影响。结果表明:①100μmol/Lt10,c12-CLA显著降低了猪皮下脂肪TG含量并提高了猪背最长肌的TG含量(P〈0.05);②100μmol/Lt10,c12-CLA显著抑制了猪皮下脂肪FAS、INSR与背最长肌HSL和LPL的基因表达,促进了猪皮下脂肪HSL和背最长肌INSR的基因表达(P〈0.05);③该研究结果从脂肪代谢关键酶类与调控激素方面揭示了t10,c12-CLA对猪皮下和背最长肌组织脂肪代谢与沉积的差异性调控机制,进一步证实了t10,c12-CLA抑制猪皮下脂肪沉积同时提高肌内脂肪含量。
In vitro tissue culture system of subcutaneous(SC)adipose and longissimus muscle taken from 30 d pig were constructed firstly,0.1 mmol/L bovine serum albumin(BSA,as control),100 μmol/L t10,c12-CLA were added to the cultures for 10 d,six replicates were performed for each treatment.At the end of 10 d treatment period,cells were harvested and stored at-70 ℃.Real-time RT-PCR and kit assay were used to characterize the effects of t10,c12-CLA on the lipogenic and lipolytic enzymes,regulation hormones,TG content in longissimus muscle and SC adipose of pig.The results showed that:the mRNA abundance of FAS and INSR,TG content in SC adipose tissue and the mRNA abundance of HSL and LPL in longissimus muscle tissue were decreased by adding t10,c12-CLA(P0.05);by contrast,HSL mRNA abundance in SC adipose tissue and the mRNA abundance of INSR,TG content in longissimus muscle tissue were increased by adding t10,c12-CLA(P0.05).These results illustrated that the effects of t10,c12-CLA on lipid metabolism and deposition of SC adipose tissue differed from that on longissimus muscle tissue,and further confirmed that t10,c12-CLA inhibited the lipid deposition of SC adipose and promoted the lipid deposition of longissimus muscle in pig.