肿瘤坏死因子α对小鼠胰岛内皮细胞表达胰岛素样生长因子结合蛋白7的影响  

TNF-α induces the expression of IGFBP7 in mouse islet endothelial cells

作　　者：袁磊[1] 杨旭光[1] 王建国[1] 徐宛玲[1] 

机构地区：[1]漯河医学高等专科学校分子医学实验室,462002

出　　处：《中国糖尿病杂志》2015年第10期931-934,共4页Chinese Journal of Diabetes

基　　金：2013年度河南省教育厅科学技术研究重点项目(13B310164)

摘　　要：目的研究肿瘤坏死因子α(TNF-α)对小鼠胰岛内皮(MS1)细胞胰岛素样生长因子结合蛋白7(IGFBP7)的mRNA和蛋白水平的影响,并探讨其机制。方法采用可溶性TNF受体Ⅰ(sTNFRⅠ)(50ng/ml)和不同浓度TNF-α(0、10、50、100、200、500ng/ml)作用MS1细胞,再收集TNF-α(100ng/ml)作用MS1细胞12h后的培养液,直接或加入sTNFRⅠ(50ng/ml)后培养新鲜的MS1细胞,采用RTPCR和Western blot检测各组MS1细胞中IGFBP7的mRNA和蛋白水平。结果与0ng/ml TNF-α组相比,100、200、500ng/ml TNF-α均可使MS1细胞中IGFBP7mRNA和蛋白水平升高(P<0.05),其中500ng/ml TNF-α组最高(P<0.05);TNF-α(100ng/ml)作用MS1细胞12h后的培养液可明显提高IGFBP7的mRNA和蛋白水平(P<0.05),但该效应可被sTNFRⅠ(50ng/ml)完全抑制(P<0.05)。结论TNF-α可能是在转录水平促进MS1细胞IGFBP7表达的。Objective To investigate the expression of mRNA and protein of insulin-like growth factor binding protein 7（IGFBP7）in mouse islet endothelial MS1 cells treated with TNF-α. MethodsMS1 cells were treated with different concentrations of TNF-α（0,10,50,100,200,500ng/ml respectively）and sTNFRⅠ（50ng/ml）.MS1 cells were exposed with TNF-α（100ng/ml）for 12 h,and then the conditioned medium was harvested and added to fresh MS1 cells in the presence or absence of soluble tumor necrosis factor receptorⅠ（sTNFRⅠ）（50ng/ml）.The mRNA level of IGFBP7 was detected by real-time PCR,and the protein level of IGFBP7 was measured by western blot. Results Compared with 0ng/ml TNF-αgroup,the expression of IGFBP7 was up-regulated in MS1 cells with 100,200 and 500ng/ml TNF-αgroups respectively（P〈0.05）and was increased most significantly in 500ng/ml TNF-αgroup（P〈0.05）.We incubated MS1 cells for 24 hin conditioned medium harvested from cells exposed to TNF-α（100ng/ml）for 12 h.The expected increase in IGFBP7 expression occurred but was completely blocked when the conditioned medium was preincubated in the presence of sTNFRⅠ（P〈0.05）.Conclusion Up-regulation of IGFBP7 is transcriptionally mediated by TNF-α.

关 键 词：肿瘤坏死因子Α 胰岛素样生长因子结合蛋白7 可溶性TNF受体Ⅰ 小鼠胰岛内皮细胞 

分 类 号：R346[医药卫生—基础医学]