肥大细胞通过细胞紧密接触对肺成纤维细胞的增殖、转化及功能的影响  

The effect of mast cell on rat pulmonary fibroblast proliferation, differentiation and function via di- rect contact

作　　者：程真顺[1] 谭维军[1] 杨炯[1] 叶燕青[1] 江平[2] 

机构地区：[1]武汉大学中南医院呼吸内科,430071 [2]武汉大学中南医院普外科科,430071

出　　处：《中华实验外科杂志》2013年第8期1598-1600,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省自然科学基金资助项目（2008CDB161）

摘　　要：目的通过肥大细胞与肺成纤维细胞接触和非接触共体培养，观察肥大细胞对肺成纤维细胞增殖、转化及功能的影响。方法分离大鼠肺成纤维细胞和腹腔肥大细胞，实验分为3组：接触共育组、非接触共育组、对照组。计数各组肺成纤维细胞数量，通过免疫细胞化学方法检测肌成纤维细胞特异性标志物α-平滑肌肌动蛋白（α-SMA），噻唑蓝（MTT）比色法测定肺成纤维细胞增殖率，通过酶联免疫吸附试验（ELISA）法检测培养液中I型胶原蛋白含量。结果各组肺成纤维细胞数目在培养的第5天均达峰值，分别为接触共育组（3．70±0．83）×10^6个／孔；非接触共育组（1．40±0．32）×10^6个／孔；对照组（1．10±0．23）×10^6个／孔，接触共育组与非接触共育组和对照组比较，差异有统计学意义（P〈0．05）；MTT检测各组肺成纤维细胞吸光度（A）值：接触共育组为0．2520±0．0318，非接触共育组为0．1250±0．0212，对照组为0．1210±0．0124，接触共育组与非接触共育组和对照组比较，差异有统计学意义（P〈0．05）；各组α-SMA染色的平均吸光度（IA）值分别为0．3190±0．0436、0．1840±0．0190、0．1730±0．0226，接触共育组与非接触共育组和对照组比较，差异有统计学意义（P〈0．05）；接触共育组，培养液中Ⅰ型胶原蛋白含量明显高于非接触共育组和对照组[（192．56±2．37）、（143．58±2．07）、（121．43±1．98）μg／L，P〈0．05）。结论肥大细胞可能通过紧密接触来促进肺成纤维细胞的增殖、转化及胶原合成。Objective To investigate the possible roles of mast cells on the proliferation and trans- formation of lung fibroblast cells by contact and non-contact co-culture of the two cell lines. Methods Fi- broblast cells were separated from adult rat lung and cultured in vitro with trypsinization. Collect rat perito- neal lavage solution to separate and purify mast cells with mast cell separating medium under asepsis cir- cumstance, then qualificated with aniline blue. Cells were divided into 3 groups, each with three sub- groups： control group; contact co-culture group; non-contact co-cuhure group. Each group were cultured for five days, counting lung fibroblast cells under microscope everyday to set up cell growth curve. After five days, α-smooth muscle actin （α-SMA） was measured by immunocy toehemistry methods in myofibro- blasts, lung fibroblast cell hyperplasia rate was measured with methyl thiazol tetrazolium （MTr） colorimet- ric method and cells count, collagen I protein in culture were measured with enzyme linked immunosorbent assay （ELISA） method. Results The difference of Fb amount between contact co-culture group and other two groups were significant [（3.70±0.83） ×10^6, （1.40±0.32） ×10^6, （1.10±0.23） ×10^6; P〈 0. 05 ） ; Contact co-culture group enhance the generation of lung fibroblast cells compared with control group and non-contact co-culture group （A： 0. 2520 ± 0. 0318, 0. 1250 ± 0. 0212, 0. 1210 ± 0. 0124; P 〈 0.05 ） ; Compared with non-contact co-culture group and control group, the average optical value （IA） of massive ct-SMA positive ceils in contact co-culture group increased significantly （0. 3190 ±0. 0436, 0. 1840 ±0. 0190, 0. 1730 ±0. 0226; P 〈0. 05） ; Compared with control group and non-contact co-culture group, collagenⅠ protein were significantly higher in contact co-culture group [ （192. 56 ± 2. 37 ）, （143.58 ±2.07）, （121.43 ±1.98） μg/L; P〈0.05]. Conclusion Mast cells can upregulate the syn- thesis of collagen I p

关 键 词：肥大细胞 肺成纤维细胞 增殖 转化 

分 类 号：R363[医药卫生—病理学]