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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

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

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【作者】 娄雅欣刘雅楠宋泉声张颖妹

【Author】 LOU Ya-xin,LIU Ya-nan,SONG Quan-sheng, ZHANG Ying-mei Peking University Center for Human Disease Genomics, Beijing 100083, China Peking University Medical and Healthy Analytical Center

【机构】 北京大学人类疾病基因研究中心北京大学人类疾病基因研究中心 北京100083北京大学医药卫生分析中心北京100083

【摘要】 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(aortic smooth 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。

【Abstract】 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 pcD-NA3. 1/rCklf1 transfected 293T cells had chemotactic effects on rat ASMC, which could be inhibited by pertussis toxin at the concentration of 10 μg/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:rCklfl has mitogenic and GiPCR-dependent chemotactic effects on rat ASMC, indicating that rCklf1 may be involved in the pathological process of atherosclerosis.

【基金】 国家高技术研究发展计划863重点项目基金(2003AA215022);高等学校全国优秀博士论文作者专项基金(200257)资助~~
  • 【文献出处】 北京大学学报(医学版) ,Journal of Peking University(Health Sciences) , 编辑部邮箱 ,2006年02期
  • 【分类号】R33
  • 【被引频次】6
  • 【下载频次】129
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