肿瘤坏死因子-α 对 THP-1 巨噬细胞中酰基辅酶 A：胆固醇酰基转移酶活性及 ACAT1 基因表达的影响
Effect of tumor necrosis factor-alpha on acyl coenzyme A: cholesteryl acyltransferase activity and ACAT1 gene expression in THP-1 macrophages.
作者信息
He Ping, Cheng Bei, Wang Yi, Wang Hongxing
机构信息
Department of Gerontology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
出版信息
J Huazhong Univ Sci Technolog Med Sci. 2007 Apr;27(2):170-2. doi: 10.1007/s11596-007-0216-9.
In order to explore the effect and mechanisms of tumor necrosis factor-alpha (TNF-alpha) on the activity of the acyl coenzyme A: cholesteryl acyltransferase (ACAT), THP-1 monocytes were cultured and induced to differentiate into macrophages with phorbol ester. TNF-alpha (60 ng/mL) was added at different time points into the macrophage-containing medium and the ACAT enzyme activity was measured by quantifying the incorporation of [1-(14)C] oleoyl CoA into cholesteryl esters. The expression of ACAT-1 protein and mRNA was respectively detected by Western blotting and RT-PCR in THP-1 macrophages 24 h after treatment with TNF-alpha (60 ng/mL). The results indicated that ACAT activity in THP-1 macrophages treated with TNF-alpha was increased in a time-dependent manner. The expression levels of ACAT-1 protein and mRNA were significantly increased in THP-1 macrophages after treatment with TNF-alpha (P<0.05). It was suggested that TNF-alpha could increase the activity of ACAT in THP-1 macrophages by up-regulating the expression of ACAT-1 gene.
为了探讨肿瘤坏死因子-α（TNF-α）对酰基辅酶A：胆固醇酰基转移酶（ACAT）活性的影响及其机制，培养THP-1单核细胞并用佛波酯诱导其分化为巨噬细胞。在不同时间点向含巨噬细胞的培养基中加入TNF-α（60 ng/mL），通过定量[1-(14)C]油酰辅酶A掺入胆固醇酯的量来测定ACAT酶活性。在用TNF-α（60 ng/mL）处理24小时后的THP-1巨噬细胞中，分别通过蛋白质印迹法和逆转录-聚合酶链反应（RT-PCR）检测ACAT-1蛋白和mRNA的表达。结果表明，用TNF-α处理的THP-1巨噬细胞中的ACAT活性呈时间依赖性增加。用TNF-α处理后，THP-1巨噬细胞中ACAT-1蛋白和mRNA的表达水平显著增加（P<0.05）。提示TNF-α可通过上调ACAT-1基因的表达来增加THP-1巨噬细胞中ACAT的活性。
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