纤维蛋白(原)及其降解产物对共培养血管平滑肌细胞生长行为学的影响

Effect of Fibrinogen and Fibrin(ogen) Degradation Products on the Growth Behavior of Vascular Smooth Muscle Cells in Coculture

目的探讨不同浓度水平的纤维蛋白原及其降解产物对共培养血管平滑肌细胞增殖与移行的影响。方法以Transwell膜为载体,建立原代培养的兔主动脉内皮细胞与平滑肌细胞的共培养体系。应用不同浓度的（0g/L、0.5g/L、1.5g/L、3.0g/L、4.5g/L和6.0g/L）纤维蛋白原及其降解产物干预共培养体系,观察共培养体系中平滑肌细胞的蛋白合成（bicinchoninic acid,BCA法）与增殖细胞核抗原（蛋白免疫印迹法）的表达,通过Transwell膜装置和细胞刮伤模型观察平滑肌细胞的趋化和迁移。结果较高浓度的纤维蛋白原（≥3.0g/L）可明显促进平滑肌细胞的蛋白合成、增殖细胞核抗原表达以及趋化迁移;纤维蛋白从0.5g/L起就可明显促进平滑肌细胞的蛋白合成。1.5～4.5g/L的纤维蛋白可明显上调增殖细胞核抗原表达,浓度过高的纤维蛋白（6.0g/L）反而抑制其表达。3.0～6.0g/L的纤维蛋白显著促进平滑肌细胞趋化,从1.5g/L起纤维蛋白就可明显加强平滑肌细胞的迁移过程;在0.5～6.0g/L之间,纤维蛋白降解产物呈浓度依赖性地促进平滑肌细胞蛋白合成、增殖细胞核抗原表达和移行。结论一定浓度的纤维蛋白原及其降解产物可以促进平滑肌细胞增殖，影响其趋化迁移过程，从而参与了动脉粥样硬化的发生发展。

Objective To investigate the effect of fibrinogen（Fg） , Fibrin（Fb） and Fibrin（ogen） degradation products（FDPs） in different concentration on the proliferation and migration of vascular smooth muscle cells（SMCs） in coculture. Methods The rabbit aorta endothelial cells（ECs）/SMCs coeulture system on Transwell membranes was established. Using different concentration of Fg, Fb and FDPs were used to interfere in the coculture system. The protein synthesis and the expression of proliferating cell nuclear antigen were observed by means of bicinchoninic aeid（BCA） method and Western blotting respectively. Migration assays were performed using the wounding model and the transwell cell culture apparatus. Results The higher concentration of Fg（≥3.0g/L） could obviously promote the expression of proliferating cell nuclear antigen（PCNA）, the protein synthesis and migration of SMCs; Fb could enhance the protein synthesis of SMCs from 0.Sg/L. Fb can obviously up-regulate the expression of PCNA dose-dependently in the range between 1.5g/L to 4.5g/L, on the contrary, the exorbitant concentration of Fb could inhibit that. Fb stimulated the ehemotaxis of SMCs dose-dependently in the range between 3.0g/L to 6.0g/L, and from 1.Sg/L, Fb obviously enhance the migration of SMCs. FDPs promote the expression of PCNA, the protein synthesis and migration of SMCs dose- dependently in the range between 0.5g/L to 6.0g/L. Conclusion Fg, Fb and FDPs, in some range of concentration, could promote the proliferation of SMCs and influence on the process of chemotaxis and migration, which involved in the pathogenesis of atherosclerosis.