TNF-α对未分化及成熟3T3-L1脂肪细胞中TSG-6基因表达的调节作用
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国家自然科学基金资助项目(39870362),江苏省教委自然科学基金资助项目(98KJB320002),江苏省医学重点人才资助项目(RC2002061)


Regulative role of tumor necrosis factor-α on TSG-6 gene expression in both undifferentiated and matured 3T3-L1 adipocytes
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    摘要:

    目的:探讨TNF-α对未分化及成熟3T3-L1脂肪细胞中TSG-6基因(tumor necrosis factor alpha-stimulated gene-6)表达水平的调节作用.方法:体外培养3T3-L1脂肪细胞,在3T3-L1细胞生长至完全融合后2天(第0天)或细胞诱导分化的第10天,应用重组TNF-α(0.1、1.0、10.0 ng/ml)干预未分化(第0天)或已分化成熟(第10天)的脂肪细胞,采用RT-PCR技术检测干预后不同时间(0、0.5、2.0、6.0、12.0、24.0 h)脂肪细胞中TSG-6基因的表达水平.结果:①不同浓度TNF-α对未分化脂肪细胞中TSG-6基因的表达水平均具抑制作用.TNF-α浓度0.1 ng/ml时,TSG-6基因表达水平2 h内下降67.30%,24 h时下降76.05%;TNF-α浓度1.0 ng/ml时,TSG-6基因表达水平0.5 h内下降52.79%,24 h时下降99.20%;TNF-α浓度10.0 ng/ml时,TSG-6基因表达水平0.5 h内下降96.14%,其表达几乎被完全抑制;②不同浓度TNF-α均能显著抑制成熟脂肪细胞中TSG-6基因的表达.TNF-α浓度0.1 ng/ml时,TSG-6基因表达水平6 h内下降33.73%,12 h内下降97.39%;TNF-α浓度1.0 ng/ml时,TSG-6基因表达水平6 h内下降78.68%,并持续至24 h;TNF-o浓度10.0 ng/ml时,TSG-6基因表达水平2 h内下降96.27%,TSG-6基因的表达几乎被完全抑制.结论:TNF-α对脂肪细胞中TSG-6基因表达具有抑制作用,其抑制效应总体趋势上呈现剂量和时间依赖性特征.

    Abstract:

    Objective:To explore the regulative role of tumor necrosis factor-a(TNF-a)on tumor necrosis factor alpha-stimulated gene-6(TSG-6)gene expression in both undifferentiated and matured 3T3-Ll adipocytes.Methods:3T 3-Ll preadipocytes were cultured and differentiated into the matured adipocytes.Recombinant TNF-a at different co ncentrations( 0.1,1.0,10 .0n g/ml)was added to the Culture medium 2days post-confluent undifferentiated adipocytes(D ay0 )or fully differentiated adipocytes( Day1 0)for various times(0, 0.5, 2.0, 6.0, 12.0, 24.0 h). Total RNA from these adipocytes was extracted and the expression of TSG-6 mRNA was evaluated by RT-PCR.Results:}1 Treatment of un differentiated3 T3-Lla dipocytesw ithT NF-aa tdi ferentco ncentrations (0.1,1. 0,10 .0n g/ml) resulted in the significant decrease in the expression of TSG-6 mRNA. When 0.1 ng/ml TNF-a was applied,the levels of TSG-6 gene expression significantly decreased by 67.30% at 2 h, and 76.05% at 24 h. When 1.0 ng/ml TNF-a was used, the levels of TSG-6 gene expression significantly decreased by 52.79% at 0.5 h, and 99.20% at 24 h. When the cells were treated with 10.0 ng/ml TNFa,TS G-6g enee xpressionw asa lmostfu llyin hibited( 96.14% decreased)at 0 .5h .(2 )Treatment of matured3 T3-Ll adipocytes with TNF-a at different concentrations( 0.1,1.0,10 .0n g/ml)a lsor esultedi nt hes ignificantd ecreasei nt hee xpressiono fT SG-6m RNA.When the cells were treated with 0.1 ng/ml TNF-(x, the levels of TSG-6 gene expression decreased by 33.73% at 6 h, and 97.39% at 12h .W hent reatedw ith1 .0n g/mlT NF-a,th ele velso fT SG-6g enee xpressiond ecreasedb y7 8.68%a t6 h ,which sustained u ntil24 h .Whenter atedw ith1 0.0n g/mlTN F-a,th ele velso fT SG-6g enee xpressiond ecerasedb y9 6.27% at2 h a ndT SG-6gene expression was almost fully inhibited. Conclusion: TNF-a can downregulate the mRNA expression of TSG-6 in both undiferentiated and matured adipocytes. The inhibitive efe ct of TNF-a on TSG -6 gene expression is in dose and time-dependent manne r.

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费莉. TNF-α对未分化及成熟3T3-L1脂肪细胞中TSG-6基因表达的调节作用[J].南京医科大学学报(自然科学版),2005,(1):25-

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  • 收稿日期:2003-11-19
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