摘要
目的通过上调SIGIRR(single Ig IL-1R-related molecule)在人气道上皮细胞株H292中的表达,研究SIGIRR对LPS诱导的H292细胞TLR4表达的影响。方法构建SIGIRR-EGFP融和蛋白的真核表达载体,运用脂质体转染的方法转染人气道上皮细胞株H292,经LPS刺激后,ELISA检测转染前后上皮细胞TNF-α分泌水平的差异,Western blot(WB)检测TLR4表达的变化。结果构建的融和蛋白表达载体SIGIRR-EGFP经EcoRI和BamHI双酶切后,电泳显示其条带大小为4.7kb和1.23kb左右,经测序证实SIGIRR的序列与GenBank公布的人cDNA序列完全一致。SIGIRR-EGFP转染细胞与pEGFP-N1转染细胞和对照组细胞相比,TNF-α产生明显下降,而TLR4表达无明显变化。结论SIGIRR上调表达可抑制LPS诱导的H292细胞TNF-α的产生,对TLR4表达无影响,提示SIGIRR在该信号通路中起"刹车"作用可能是通过抑制其下游通路中某个或某些调节蛋白的表达完成的。
Objective To investigate the effects of SIGIRR on TLR4 of human air-way epithelial cells H292 induced by LPS. Methods The construct of a eukaryotic expression vector for single Ig IL-1R-related molecule fused with enhanced green fluorescent protein （EGFP） gene was transfected into H292 induced by LPS. Then the concentration of TNF-α was detected with ELISA and the protein of TLR4 was observed with Western blot. Results The level of TNF- α, but not TLR4, of SIGIRR-EGFP transfected H292 decreased significantly compared with untransfected and pEGFP-N1 transfected cells. Conclusion Over-expression of SIGIRR can inhibit LPS-induced inflammation through inhibiting the production of TNF-α but not TNF-α.
出处
《西部医学》
2009年第4期530-532,共3页
Medical Journal of West China
基金
国家自然科学基金资助项目(No.30600272)
