大鼠趋化素样因子1对主动脉平滑肌细胞的趋化作用和增殖促进作用

目的:探讨大鼠趋化素样因子1(rat chemokine-like factor 1,rCklf1)对大鼠主动脉平滑肌细胞(aorticsmooth muscle cells,ASMC)的趋化作用和促增殖作用.方法:将rCklf1真核细胞表达载体瞬时转染293T细胞,收获培养上清液,分析rCklf1对大鼠ASMC的趋化作用.同时,用3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑(MTT)实验分析rCklf1真核细胞表达载体瞬时转染细胞上清液以及rCklf1在细胞内瞬时超表达对ASMC增殖的影响.结果:rCklf1真核细胞表达载体瞬时转染的293T细胞培养上清液对大鼠ASMC具有趋化作用,10倍稀释的转染上清液所趋化的细胞数是对照组的2.2倍,这种趋化作用可以被浓度为10μg/L的百日咳毒素(pertussis toxin, .PTX)抑制.同时,rCklf1对ASMC有促增殖作用,10倍稀释的rCklf1真核细胞表达质粒瞬时转染上清液和rCklf1在细胞内瞬时超表达均可以促进ASMC的增殖,光密度值分别为对照组的1.3和1.5倍.结论:rCklf1对大鼠ASMC具有促增殖作用和依赖于Gi蛋白偶联受体(Gi protein coupled receptor,GiPCR)的趋化作用,可能参与动脉粥样硬化的病理过程.

Chemotactic and mitogenic effect of rat chemokine-like factor 1 on aortic smooth muscle cells

OBJECTIVE: To study the effects of rat chemokine-like factor 1 (rCklf1) on chemotaxis and proliferation of rat aortic smooth muscle cells(ASMC). METHODS: The recombinant eukaryotic expression vector pcDNA3.1/rCklf1 was transiently transfected into 293T cells. The supernatants were harvested for chemotactic and proliferation assays. pcDNA3.1/rCklf1 was also transfected into ASMC and the proliferation of transfected cells was detected by MTT assays. RESULTS: The 10 fold diluted supernatants of pcDNA3.1/rCklf1 transfected 293T cells had chemotactic effects on rat ASMC, which could be inhibited by pertussis toxin at the concentration of 10 microg/L. Furthermore, the 10 fold diluted supernatants of rCklf1 transfected 293T cells promoted the proliferation of ASMC (1.3 fold compared with the control) and transient overexpression of rCklf1 in ASMC had promoting activities on the proliferation of rat ASMC (1.5 fold compared with the control at 72 h). CONCLUSION: rCklf1 has mitogenic and GiPCR-dependent chemotactic effects on rat ASMC,indicating that rCklf1 may be involved in the pathological process of atherosclerosis.