促酰化蛋白对3T3-L1脂肪细胞糖脂代谢的影响
投稿时间:2017-05-26  修订日期:2017-06-12  点此下载全文
引用本文:刘婧,胡秀芬,卢慧玲,温宇.促酰化蛋白对3T3-L1脂肪细胞糖脂代谢的影响[J].医学研究杂志,2018,47(2):69-73
DOI: 10.11969/j.issn.1673-548X.2018.02.017
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作者单位E-mail
刘婧 430030 武汉, 武汉市儿童医院  
胡秀芬 430030 武汉, 华中科技大学附属同济医院儿科  
卢慧玲 430030 武汉, 华中科技大学附属同济医院儿科  
温宇 430030 武汉, 华中科技大学附属同济医院儿科 2721837622@qq.com 
基金项目:国家自然科学基金青年科学基金资助项目(30800385);教育部新教师基金资助项目(200804871059)
中文摘要:目的 观察胰岛素抵抗状态下3T3-L1成熟脂肪细胞促酰化蛋白(acylation stimulating protein,ASP)刺激下脂代谢关键酶(LPL、HSL、perilipin、DGAT)基因表达的影响,以及胰岛素经典信号通路(IRS-1、PI3K)蛋白表达的影响。方法 3T3L-1脂肪细胞给予不同浓度(0、0.125、0.5、1.0mmol/L)油酸及棕榈酸孵育过夜。real-time PCR方法定量检测ASP刺激下脂肪细胞LPL、HSL、perilipin、DGAT基因表达水平。Western blot法检测ASP和INS刺激下脂肪细胞HSL、perilipin、IRS-1、PI3K蛋白表达水平。结果 ASP促进脂代谢关键酶HSL、LPL、perilipin和DGAT的基因表达。其中0.5mmol/L油酸组增加5倍HSL、1.87倍LPL、6.87倍perilipin、2.5倍DGAT基因表达(P<0.01)。1.0mmol/L油酸组增加3.26倍HSL、2.98倍LPL、5.46倍perilipin、2倍DGAT基因表达(P<0.01)。0.5mmol/L棕榈酸组增加1.98倍HSL、4.63倍DGAT基因表达(P<0.05)。胰岛素和ASP刺激下显著增加脂肪酸孵育的成熟脂肪细胞HSL、perilipin、PI3K蛋白的表达。胰岛素刺激状态下油酸组增加1.75倍HSL、1.27倍perilipin、2.59倍PI3K蛋白的表达(P<0.01),棕榈酸组增加85%(P<0.05) HSL、96%(P<0.05) perilipin、3.66倍(P<0.01) PI3K蛋白的表达。ASP刺激状态下油酸组增加76%(P<0.05) HSL、86%(P<0.05) perilipin蛋白的表达,棕榈酸组增加76%(P<0.05) perilipin、1.27倍(P<0.01) PI3K蛋白表达。胰岛素及ASP刺激下脂肪细胞IRS-1蛋白表达差异无统计学意义。结论 胰岛素和ASP可增加脂代谢关键酶HSL、perilipin蛋白及经典胰岛素信号通路PI3K蛋白的表达。ASP可以从不同程度上增加糖脂代谢关键酶HSL、LPL、perilipin和DGAT基因的表达,通过增加这些酶的表达,提高酶的敏感度,参与调节脂肪细胞糖脂代谢。
中文关键词:脂肪细胞  胰岛素抵抗  促酰化蛋白  糖脂代谢关键酶
 
Effects of Acylation Stimulating Protein on the Lipometabolism in 3T3-L1 Adipocytes
Abstract:Objective To evalute the affect on secretion levels and protein expression of the key enzyme of lipid metabolism(LPL,HSL,perilipin,DGAT),and the affect on protein expression of classical signaling pathways of insulin (IRS-1, PI3K) in 3T3-L1 adipocytes with acylation stimulating protein(ASP)stimulated under the condition which produce insulin resistance by free fatty acids.Methods The different concentrations of the Oleate and Palmitate(0,0.125,0.5,1.0mmol/L) were added to cultured 3T3-L1 adipocytes overnight. The real-time-PCR were used to detected mRNA of LPL, HSL, Perilipin, DGAT. The western blot were used to detected protein of HSL, prelipin, IRS-1, PI3K.Results After incubated with ASP, comparing with that of 0mmol/L, the levels of HSL, perilipin, LPL and DGAT mRNA expression were increased. In insulin and ASP stimulating condition, comparing with the 0mmol/L,the levels of HSL, perilipin, PI3K protein expression were increased. And comparing with the insulin stimulated, the protein expression of HSL,perilipin IRS-1 and PISK failed to significantly increase with the ASP stimulated.Conclusion Under the condition of insulin resistance, ASP can increase the mRNA expression of HSL, LPL, perilipin and DGAT by increasing the expression of these enzymes to improve the sensitivity and improve the use of the body's fat cell, to regulate the glucose and lipid metabolism.INS and ASP can increase the protein expression of the key enzyme of lipid metabolism (HSL, perilipin), and the classical insulin signaling pathway (PI3K).
keywords:Adipocytes  Insulin resistance  Acylation stimulating protein  Key enzyme of lipometabolism and glycometabolism
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