摘要
目的探讨不同处理因素对呼吸道合胞病毒(RSV)感染人支气管上皮细胞16HBE表达胸腺基质淋巴细胞生成素(TSLP)的影响。方法用Hep-2细胞扩增病毒,并进行病毒毒力鉴定;试验分6组:对照组、RSV组、RSV/anti-TLR3组、RSV/IFN-γ组、RSV/IL-4组、RSV/地塞米松组,感染复数(MOI)为2的RSV病毒液吸附感染1h,不同处理因素分别加入培养基共培养;培养6h后用实时荧光定量RT-PCR检测各组细胞TSLPmRNA表达水平变化,培养24h后应用Western blotting检测各组细胞TSLP蛋白表达水平变化。结果经Hep-2细胞培养扩增获得足量RSV病毒;RSV感染16HBE细胞6h,可使细胞TSLPmRNA表达水平增高(是对照组的1.63±0.08倍)(P<0.001);加入anti-TLR3阻断TLR3受体,细胞TSLPmRNA表达量下降(P=0.034);加入重组人IFN-γ,细胞TSLPmRNA表达水平下降(P<0.001);加入重组人IL-4,TSLPmRNA表达水平升高(P=0.025);加入地塞米松,细胞TSLPmRNA表达明显下降(P<0.001)。RSV感染后细胞TSLP蛋白表达量升高,约为对照组的1.9倍(P<0.001),TLR3抗体阻断TLR3受体可使TSLP表达降低,但与RSV感染组比较无统计学意义(P=0.114),IFN-γ降低TSLP表达升高(P=0.020),IL-4协同RSV感染引起的TSLP表达升高(P=0.014),地塞米松显著抑制RSV感染细胞TSLP表达(P<0.001)。结论RSV感染引起人支气管上皮细胞16HBE合成TSLP增加;IFN-γ、anti-TLR3和地塞米松能抑制RSV感染引起的16HBE细胞TSLP合成,而IL-4能协同RSV感染促进TSLP合成。
Objective To investigate the effects of different factors on the expressions of thymic stromal lymphopoietin （TSLP） in respiratory syncytial virus （RSV）-infected human airway epithelial cell line 16HBE cells. Methods RSV amplified by infecting Hep-2 cells was identified for its virulence. 16HBE cells were divided into six groups, namely the control group, RSV group, RSV/anti-TLR3 group, RSV/IFN-γ group, RSV/IL-4 group and RSV/dexamethasone group with corresponding treatments. Real-time RT-PCR was used to examine the expression of TSLP mRNA in the cells 6 h after RSV infection. Western blotting was used to examine TSLP protein expression in the cells 24 h after the infection. Results The expression of TSLP mRNA in 16HBE cells 6 h after RSV infection increased by 1.63±0.08 folds as compared to the expression level in the control cells. The expression of TSLP mRNA was significantly decreased in RSV-infected cells treated with anti-TLR3 antibody （P=0.034） and recombinant human IFN-γ （P0.001）, but increased with the treatment by recombinant human IL-4 （P=0.025）. Dexamethasone significantly inhibited the expression of TSLP mRNA in RSV-infected cells （P0.001）. The production of TSLP protein in 16HBE cells increased by 1.9 folds （P0.001） 24 h after RSV infection, but underwent no significant changes after treatment with anti-TLR3 antibody （P=0.114）. Recombinant human IFN-γ significantly decreased while IL-4 enhanced the expression of TSLP protein in the infected cells （P=0.020 and 0.014, respectively）. Dexamethasone significantly inhibited the increment of TSLP protein expression in RSV-infected cells （P0.001）. Conclusions RSV infection can enhance the expressions of TSLP in human airway epithelial cells. IFN-γ, anti-TLR3 and dexamethasone can inhibit the elevation of TSLP expression induced by RSV infection, but IL-4 synergistically enhances its expression.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2010年第3期519-522,共4页
Journal of Southern Medical University
基金
中华医学会慢性呼吸道疾病专项基金(07010130021)
