The Effection Of Mast Cell On Proliferation And Transformation Of Pulmonary Fibroblast In Rat

Objective:To investigate the possible roles of mast cells on the proliferation and transformation of lung fibroblast cells by contact and non-contact co-culture of the two cell lines.Methods:Fibroblast cells were separated from adult rat lung and cultured in vitro with trypsinization. Identify cells by immunohistochemistry technique and take them for usage after three passages. Collect rat peritoneal lavage solution to separate and purify mast cells with mast cell separating medium under asepsis circumstance, then qualificated with aniline blue. Cells were divided into 3 groups, each with three subgroups:control group (lung fibroblast cell culture of normal rat); contact co-culture group (contact co-culture of mast cells and lung fibroblast cells); non-contact co-culture group (non-contact co-culture of mast cells and lung fibroblast cells, in transwell culture capsule). Each group were cultured for five days,counting lung fibroblast cells under microscope everyday to set up cell growth curve. After five days,α-MA was measured by immunocy tochemistry methods in myofibroblasts, lung fibroblast cell hyperplasia rate was measured with MTT colorimetric method and cells count, bFGF、TGFβ1 and collagen I protein in culture were measured with ELISA method.Results:①Cell counting results suggest that Fb amount in contact co-culture group were increased obviously than non-contact co-culture group and control group in every stage(P<0.05).The difference of Fb amount between contact co-culture group and other two groups were significant, but were not significant between non-contact co-culture group and control group(P>0.05).②MTT results indicate that co-culture of mast cells and lung fibroblast cells can obviously increase the generation of the latter. Contact co-culture group enhance the generation of lung fibroblast cells compared with control group and non-contact co-culture group (P<0.05). The generation of lung fibroblast cells in non-contact co-culture group were also increased compared with control group,but with no significant difference (P>0.05).③Compared with control group and non-contact co-culture group,, massiveα-SMA positive cells were detected in contact co-culture group, the average optical value (IOD) increased significantly (P<0.05). Non-contact co-culture group is increased compared with control group,but with no significant difference (P>0.05).④Compared with control group and non-contact co-culture group, bFGF、TGFβ1 and collagen I protein were higher in contact co-culture group(P<0.05). Compared with control group, non-contact group has higher protein expression, but with no significant difference(P>0.05).Conclusion:Mast cells can upregulate the synthesis of bFGF and TGFβ1 by a compact contact way with lung fibroblast cells;The proliferation and transformation of lung fibroblast cells is more significant when contact co-culture with mast cells.