转化生长因子-β_1对人成熟树突状细胞生物物理特性和迁移能力的影响

The Effects of Transforming Growth Factor-beta1 on Biophysical Characteristics and Motility of Mature Dendritic Cells

目的:研究转化生长因子β1(TGF-β1)对人成熟树突状细胞(m DCs)生物物理特性与迁移能力的影响。方法:用不同浓度TGF-β1(1μg/L、3μg/L、5μg/L、7μg/L)处理m DCs 24 h,按TGF-β1浓度将m DCs分为4组和对照组(未加入细胞因子);采用细胞渗透脆性实验检测m DCs的渗透脆性(未破碎细胞百分率),通过细胞电泳实验测定m DCs电泳率,荧光偏振实验和微吸管系统分别测定m DCs荧光片振度和被吸入管内的长度,以反映m DCs细胞膜的流动性和变形性,利用Transwell系统检测m DCs跨内皮细胞迁移百分率。结果:与对照组相比,(1)细胞渗透脆性,在1μg/L组m DCs破碎百分率增加,5μg/L组m DCs破碎百分率降低,差异有统计学意义(P<0.05);(2)细胞电泳率,1μg/L和3μg/L组m DCs电泳率下降,差异有统计学意义(P<0.05或P<0.01);(3)细胞膜流动性和细胞变形性,细胞膜流动性在5μg/L组增加;变形能力在5μg/L和7μg/L组增加,差异有统计学意义(P<0.01或P<0.05);(4)细胞迁移能力,1μg/L和3μg/L组m DCs跨内皮细胞迁移率下降,差异有统计学意义(P<0.05或P<0.01)。结论:TGF-β1通过影响m DCs的生物物理特性从而抑制其跨内皮细胞迁移的能力。

Objective：To study the effects of TGF-β1 on the biophysical characteristics and motility of mDCs.Methods：mDCs were treated with TGF-β1 at different concentrations （0,1,3,5,7 μg/L,respectively） for 24 hours,mDCs were divided into five groups （1 μg/L group,3 μg/L group,5 μg/L group,7 μg/L group and control group without TGF-β1） ;the percentage of non-broken cells was determined by osmotic fragility test under different osmotic pressure to test osmotic fragility of mDCs ; the electrophoretic mobility of mDCs was measured by cell electrophoresis assay; fluorescence sheet was determined by fluorescence polarization experiment ; inhalation tube length of mDCs was determined by micro straw system; the percentage of mDCs transendothelial migration was determined by Transwell system.Results：Compared with those of control group：（1） the osmotic fragility of cells,the percentage of non-broken cells was increased at 1 μg/L group but decreased at 5 μg/L group （P ＜ 0.05） ; （2） The electrophoretic mobility （EPM） of mDCs were decreased in 1 μg/L and 3 μg/L group,differences were statistical significant （P ＜ 0.05 and P ＜ 0.01）.（3） The membrane fluidity of mDCs was significantly increased in 5 μg/L group; the deformation abilities of cells were significantly increased in 5 and 7 μg/L group,differences were statistical significant （P ＜ 0.01 and P ＜ 0.05）.（4） The transendothelial migration abilities of mDCs were significantly decreased in 1 μg/L and 3 μg/L group （P ＜ 0.05 and P ＜ 0.01）.Conclusions：TGF-β1 can inhibit transendothelial migration abilities of mDCs through affecting biophysical characteristics of mDCs.