摘要
采用RTPCR、MTT比色法和油红O染色提取法,分别测定外源TNFα、胰岛素对原代培养大鼠前体脂肪细胞增殖及脂肪生成和碳水化合物反应元件结合蛋白(ChREBP)、固醇调控元件结合蛋白(SREBP)1c、脂肪酸合酶(FAS)及乙酰辅酶A羧化酶(ACC)1基因转录表达的影响。结果表明,TNFα可有效抑制胰岛素对原代培养大鼠前体脂肪细胞增殖的促进作用,并通过抑制胰岛素对转录因子SREBP1c和脂肪酸合酶FAS及ACC1基因转录表达的促进作用,减少脂肪酸的生物合成,抑制成熟脂肪细胞的脂肪生成,证明TNFα是动物脂肪形成的有效抑制因子。此外,TNFα下调ChREBPmRNA表达。
TNF-α is a pleiotropic cytokine secreted in adipose tissue. To study the effects of TNF-α on adipogenesis stimulated by insulin in vitro, rat preadipocytes were isolated by collagenase treatment of adipose tissue from male SD rats （20 d-old）, and cultured in media with TNF-α, insulin and TNF-α ＋ insulin, respectively. Preadipocyte proliferation was measured with MTT. The lipogenesis and relative levels of lipogenic-related gene mRNAs in mature adipocytes were measured through Oil Red O staining and semisuantitative RT-PCR, respectively. The results showed that TNF-α effectively inhibited preadipocyte proliferation stimulated by insulin and decreased lipogenesis through inhibiting stimulation of insulin on transcriptional expressions of lipogenic genes such as acetyl-CoA carboxylase α （ACC1）, fatty acid synthase （FAS）, sterol regulatory element binding protein （SREBP） -1c in adipocytes, suggesting that TNF-α is a effective inhibitor of adipogenesis in adipose tissue. In addition, mRNA expression of the carbohydrate response element binding protein （ChREBP） was significantly inhibited by TNF-α, while it was not influenced by insulin in adipocytes in low glucose media [Acta Zoologica Sinica 52 （1） ： 123- 129, 2006].
出处
《动物学报》
SCIE
CAS
CSCD
北大核心
2006年第1期123-129,共7页
ACTA ZOOLOGICA SINICA
基金
国家重点基础研究发展计划(973)项目(No.2004CB117506)~~
