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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 effects of tumor necrosis factor α (TNF-α) on the mRNA and protein level of insulin-like growth factor binding protein 7 (IGFBP7) in mouse pancreatic islet endothelium (MS1) cells and to explore its mechanism. Methods MS1 cells were treated with soluble TNF receptor Ⅰ (50ng / ml) and different concentrations of TNF-α (0, 10,50,100,200,500ng / ml) After MSCs were treated with sTNFRI (50ng / ml) for 12h, fresh MS1 cells were cultured. The mRNA and protein levels of IGFBP7 in MS1 cells were detected by RTPCR and Western blot. Results Compared with 0 ng / ml TNF-α group, 100, 200 and 500 ng / ml TNF-α significantly increased the mRNA and protein levels of IGFBP7 in MS1 cells (P <0.05), of which 500 ng / ml TNF- (P <0.05). The effect of TNF-α (100ng / ml) on MS1 cells for 12h significantly increased IGFBP7 mRNA and protein levels (P <0.05), but this effect was completely inhibited by sTNFRI (50ng / ml) P <0.05). Conclusion TNF-α may promote the expression of IGFBP7 in MS1 cells at transcriptional level.