ET-1和bFGF促进大鼠血管平滑肌细胞增殖的协同作用及其机制研究

摘要

AIM: The synergistic effect of basic fibroblast growth factor (bFGF) and endothelin- 1 (ET-1 ) on rat aortic vascular smooth muscle cells(VSMC) proliferation was observed. The possible mechanism of the synergism was also investigated. METHODS: BrdU incorporation and cell counting method were adopted to value the pro - proliferative effect of VSMC. Western- blotting was used to observe the variation of bFGF and FGFR - 1 isoforms expression. RESULTS: bFGF and ET- 1 could promote VSMC proliferation separately, and synergistically in combination. The synergism was dose - and time - dependent. ET- 1 increased all the three bFGF isoforms and FGFR - 1 protein level in dose - and time- dependent manner. In addition, after exhaustion of intracellular PKC, the upregulation effects of ET- 1 on bFGF and FGFR - 1 expressions in VSMC both inclined. CONCLUSION: bFGF and ET- 1 had synergistic effect on VSMC proliferation. ET- 1 may increase the responsiveness of VSMC to bFGF through modulation of bFGF isoforms together with FGFR - 1, which was PKC - dependent.%目的：观察碱性成纤维细胞生长因子(bFGF)和内皮素-1(ET-1)促进血管平滑肌细胞(VSMC)增殖的协同作用，并进一步研究可能的机制。方法：5-溴-2'-脱氧尿嘧啶(BrdU)掺入法和细胞计数法观察对VsMC增殖的影响，westem免疫印迹法观察ET-1对VSMC bFCF和成纤维细胞生长因子受体-1(FGFR-1)蛋白表达的影响。结果：bFGF和ET-1能协同促进VsMC BrdU掺入和细胞增多，并且在一定剂量和时间范围内呈量效、时效关系。同时ET-1剂量依赖性上调bFGF和FGFR-1蛋白，表达高峰分别为24和48 h，二丁酸佛波脂(PDBU)预耗竭细胞内蛋白激酶C(PKC)后该上调作用显著下降。结论：bFGF和ET-1对VSMC增殖具有协同作用，与ET-1上调bFGF和FGFR-1蛋白有关，上调作用呈PKC依赖性。