BAMBI通过促进ERK1/2磷酸化抑制猪前体脂肪细胞分化

BAMBI inhibits porcine preadipocyte differentiation by facilitating ERK1/2 phosphorylation

为了研究BAMBI在猪前体脂肪细胞分化过程中的作用,构建了BAMBI慢病毒干扰载体,包装并感染猪前体脂肪细胞,采用油红O染色、油红O提取比色法检测猪前体脂肪细胞分化情况,采用Real-time qPCR、Western blotting检测成脂标志基因mRNA以及蛋白水平表达的变化情况。结果表明,BAMBI慢病毒干扰载体感染前体脂肪细胞后显著降低了BAMBI的表达,shRNA2干扰效率最高,达到了60%以上,干扰BAMBI后能增加猪脂肪细胞的脂质积累,增加了成脂标志基因过氧化物酶体增殖物激活受体γ(Peroxisome proliferator-activated receptorγ,PPARγ)和脂肪酸结合蛋白2(Adipocyte protein 2,ap2)的表达。此外,干扰BAMBI后ERK1/2的磷酸化水平减少了。这些结果表明,BAMBI可能通过促进ERK1/2的磷酸化抑制脂肪细胞分化。

To study the role of BAMBI in adipogenesis, we constructed lentivirus interfering vector targeting on porcine BAMBI, packaged and infected the porcine preadipocyte. The differentiation state of preadipocyte was detected by Oil Red O staining and Oil Red O extraction assay and the expression levels of adipogenic marker genes were detected by Real-time qPCR and Werstern bloting. Results show that BAMBI expression was significant decreased after lentivirus infection, which was repressed more than 60% by shRNA2. Moreover, knockdown BAMBI increased the lipid accumulation of porcine preadipocyte and improved the expression of PPARy （peroxisome proliferator-activated receptorT） and ap2 （adipocyte protein 2）. In summary, these data indicated that BAMBI inhibited adipocyte differentiation by facilitating the phosphorylation of ERK 1/2.