hTIM-3对经屋尘螨或/和地塞米松处理16HBE细胞株炎症相关因子表达的影响

The effect of hTIM-3 on expression of inflammation-related factors in 16HBE cells treated with a house dust mite extract or/and dexamethasone

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 investigate the effect of hTIM-3on expression of inflammation-related factors in 16 HBE cells treated with a house dust mite extract or/and dexamethasone. Methods 16 HBE cells transfected with pEGFP-C2-hTIM-3and 16 HBE cells transfected with pEGFP-C2 were treated with different concentrations of a house dust mite extract or/and dexamethasone.The mRNA of cells was extracted with TRIzol and reverse transcribed into cDNA.Realtime quantitative PCR was used to detect the expression of hTIM-3,IFN-γ,TNF-α,and GATA-3mRNA in cells treated with the extract or/and dexamethasone after transfection.The effect of hTIM-3on expression of inflammation-related factors in 16 HBE cells treated with a house dust mite extract or/and dexamethasone was analyzed and discussed along with the rationale for that effect. Results When 16 HBE cells were treated with a house dust mite extract or/and dexamethasone,expression of hTIM-3（P〈0.01）,IFN-γ（P〈0.01）,and TNF-α（P〈0.01）in 16 HBE cells transfected with a pEGFP-C2-hTIM-3plasmid differed markedly from expression in 16 HBE cells transfected with pEGFP-C2.Expression of GATA-3did not change substantially（P〉0.05）.Furthermore,expression of hTIM-3was correlated with expression of IFN-γ,while expression of hTIM-3was inversely correlated with expression of TNF-αin 16 HBE cells transfected with the pEGFP-C2-hTIM-3plasmid.Expression of hTIM-3 was not correlated with expression of TNF-αin 16 HBE cells transfected with pEGFP-C2. Conclusion Upregulation of hTIM-3may increase the expression of IFN-γand downregulate expression of TNF-αin 16 HBE cells.