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目的研究T细胞免疫球蛋白及粘蛋白域蛋白3(简称TIM-3)对人气道上皮细胞株16HBE经屋尘螨或/和地塞米松处理后炎症相关因子表达的调控。方法实验分两组,分别为转染pEGFP-C2和pEGFP-C2-hTIM-3的16HBE细胞株。两组细胞株分别用一定浓度的屋尘螨或/和地塞米松处理。提取细胞mRNA并逆转录为cDNA采用实时荧光定量PCR检测转染细胞hTIM-3、IFN-γ、TNF-α和GATA-3mRNA表达水平;分析hTIM-3对经屋尘螨或/和地塞米松处理的16HBE细胞株炎症相关细胞因子表达的调控及其机制。结果实时荧光定量PCR显示,屋尘螨或/和地塞米松处理,转染pEGFP-C2-hTIM3质粒的16HBE细胞株TIM-3、IFN-γ、TNF-αmRNA的表达与相应处理的转染pEGFP-C2质粒16HBE细胞比较差异均有统计学意义(P<0.01),但GATA-3mRNA表达无明显变化(P>0.05)。在转染pEGFP-C2-hTIM3质粒的16HBE细胞株,TIM-3 mRNA的表达与IFN-γmRNA的表达呈正相关,与TNF-αmRNA的表达呈负相关,而在转染pEGFP-C2质粒的16HBE细胞未存在相关性。结论在16HBE细胞中,TIM-3的上调可能上调IFN-γ的表达,下调TNF-α的表达。
Objective To study the regulation of inflammation-related factor expression by T cell immunoglobulin and mucin domain protein 3 (TIM-3) on human airway epithelial cell line 16HBE after house dust mite and / or dexamethasone treatment. Methods The experiment was divided into two groups: 16HBE cell line transfected with pEGFP-C2 and pEGFP-C2-hTIM-3 respectively. Two groups of cell lines were treated with house dust mites and / or dexamethasone at a certain concentration. The expression of hTIM-3, IFN-γ, TNF-α and GATA-3 mRNA in transfected cells was detected by real-time fluorescence quantitative PCR. The expression of hTIM-3 mRNA in hADM- Regulation of the expression of inflammatory cytokines in 16HBE cell line and its mechanism. Results Real-time PCR showed that the expression of TIM-3, IFN-γ and TNF-α mRNA in 16HBE cells transfected with pEGFP-C2-hTIM3 plasmid was significantly inhibited by house dust mite and / or dexamethasone compared with the correspondingly transfected pEGFP -C2 plasmid 16HBE cells were significantly different (P <0.01), but GATA-3 mRNA expression had no significant change (P> 0.05). In 16HBE cells transfected with pEGFP-C2-hTIM3 plasmid, the expression of TIM-3 mRNA was positively correlated with the expression of IFN-γmRNA and negatively correlated with the expression of TNF-αmRNA. In 16HBE cells transfected with pEGFP-C2 plasmid, No correlation exists. Conclusion In 16HBE cells, upregulation of TIM-3 may up-regulate the expression of IFN-γ and down-regulate the expression of TNF-α.