肥大细胞对肺成纤维细胞生长及胶原合成的影响

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,了解肥大细胞对肺成纤维细胞生长及胶原合成的影响。方法:分离成年大鼠肺成纤维细胞和腹腔肥大细胞,实验分为三组:对照组,正常大鼠肺成纤维细胞培养;接触共育组,肥大细胞与肺成纤维细胞接触共体培养;非接触共育组,肥大细胞与肺成纤维细胞非接触共体培养。每组设3复孔,并作细胞爬片,共培养5 d,每日镜下计数肺成纤维细胞数量,建立生长曲线,5 d后用MTT比色法测定肺成纤维细胞增殖率,通过ELISA法检测培养液中Ⅰ型胶原蛋白含量。结果:①细胞计数显示:接触共育组各时段成纤维细胞的数目均较非接触共育组和对照组明显增多。接触共育组中成纤维细胞增殖数与非接触共育组和对照组比较,有明显差异(P<0.05);非接触共育组和对照组比较,差异不明显(P>0.05)。②MTT结果:接触共育组肺成纤维细胞光密度值明显高于对照组和非接触共育组(P<0.05),提示接触共育组肺成纤维细胞增殖明显高于对照组和非接触共育组,非接触共育组比对照组也高,但差异不明显(P>0.05)。③ELISA法检测结果提示,在肥大细胞与肺成纤维细胞接触共育培养组,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(P<0.05),提示接触共育组肺成纤维细胞合成Ⅰ型胶原蛋白明显增加,非接触共育组比对照组也高,但差异不明显(P>0.05)。结论:肥大细胞影响肺成纤维细胞的增殖及胶原合成;肥大细胞可能通过紧密接触来促进肺成纤维细胞的增殖及胶原合成。

Effect of Mast Cell on Proliferation of Fibroblast and Synthesis of Collagen

Objective:To investigate the possible roles of mast cells on the proliferation and synthesis of collagen by contact and non-contact co-culture of two cell lines.Methods:Fibroblasts(Fb) were separated from adult rat lung and cultured in vitro with trypsinization.Rat peritoneal lavage solution was collected to separate and purify mast cells with mast cell separating medium under asepsis circumstance.Cells were divided into 3 groups:control group(lung fibroblast cell culture of normal rat);contact co-culture group,and non-contact co-culture group(in transwell culture capsule).Each group were cultured for five days,and lung fibroblast cells were counted under microscope everyday to set up cell growth curve.After five days,lung fibroblast cells' proliferation was measured with MTT colorimetric method and cell counts,while collagen I protein ex- pression were measured with ELISA method.Results;①Cell counting results suggested that Fb amount in contact co-culture group were increased obviously than in non-contact co-culture group and control group in every stage(F<0.05).But no significant difference in Fb amount was found between non-contact co-culture group and control group(P>0.05).②MTT results indicated that co-culture of mast cells and lung fibroblast cells could obviously increase the generation of the latter.Contact co-culture group enhanced the generation of lung fibroblast cells as compared with control group and non-contact co-culture group(P<0.05).③Compared with that in control group and non-contact co-culture group,collagen I protein were higher in contact co-culture group(P<0.05),while no significant difference was found between control group and noncontact culture group(P>0.05).Conclusion;Mast cells can up-regulate the proliferation of lung fibroblast cells and the synthesis of collagen by a compact contact way with lung fibroblast cells.