文档介绍：聚肌胞对人早孕绒毛外滋养细胞TLR3表达的调节作用
【摘要】目的通过体外实验研究Toll样受体3(TLR3)在人早孕绒毛外滋养细胞株TEV-1的表达及聚肌胞PolyI:C对TLR3 mRNA和蛋白的调节作用。方法体外培养TEV-1细胞;用不同时间及浓度聚肌胞刺激TEV-1细胞后RT-PCR和流式细胞仪检测其TLR3 mRNA和蛋白的表达变化;免疫细胞化学SP染色法检测TLR3蛋白的定位表达。结果(1)PolyI:C 刺激可上调TEV-1细胞株TLR3 mRNA的表达(P<),且与刺激时间和剂量呈正相关。(2) PolyI:C 刺激可上调TEV-1细胞株TLR3蛋白表达的平均荧光强度(P<);但荧光阳性细胞数刺激12h与刺激前比较差异无显著性(P>);刺激24h、48h的荧光阳性细胞数与刺激时间和剂量呈正关(24h P<;48h P<)。(3)免疫细胞化学显示TLR3主要定位于TEV-1细胞株的细胞浆。结论 TEV-1细胞株TLR3的mRNA及蛋白的表达与Poly(I:C) 刺激时间及剂量呈正相关;TLR3主要定位于TEV-1细胞株的细胞浆,而细胞核及细胞膜没有观测到明显表达。提示人早孕绒毛外滋养细胞可能通过细胞浆的TLR3参与早孕母-胎界面的免疫反应。
【关键词】 TLR3;Poly(I:C);TEV-1细胞株;流式细胞仪;平均荧光强度;荧光阳性细胞数;免疫细胞化学
[Abstract] Objective To investigate the regulatory effect of PolyI:C on the expression of TLR3 mRNA and protein in the human extravillous trophoblast cell line (TEV-1),and whether the regulatory effect depends on PolyI:C’s dose and Semi-quantitative RT-PCR analysis,flow cytometry and immuo-cytochemistry were used to dedect TLR3 mRNA and protein expression level and location in TEV-I cell (1)The expression of TLR3 mRNA in TEV-I cell line significantly increased with the PolyI:C’s stimulation time and dose increase; compare to without PolyI:C’s stimulation(P<).The mean fluorescence intensity of the TLR3 protein in TEV-I cell line increased with the PolyI:C’s stimulation time and dose increase; compare to without PolyI:C’s stimulation(P<).(2)The number of fluorescence positive cell of the TLR3 protein stimulated 12 hour had no statistics significance(P>); But its increased with the stimulation time and dose increase in 24 and 48 hour intervention(24h P< ; 48h P<).(3)The Immuocytochemistry result revealed that TLR3 protein was expressed in TEV-I cell line’s ,it’s hardly to see the expression in the cellular membrane and TLR3 protein was expressed in TEV-I cell line and localized to the increase of mRNA and protein expression of TLR3 in T