摘要
目的探讨重组人CD40配体及环氧合酶抑制剂对人Ⅰ型胶原α2(Ⅰ)链启动子活性的作用。方法体外培养人血管平滑肌细胞,人Ⅰ型胶原α2(Ⅰ)链基因启动子重组质粒pCOLH22.4和pCOLH21.6扩增和酶切鉴定后,通过脂质体转染血管平滑肌细胞,重组人CD40配体刺激转染后细胞,酶联免疫吸附法检测CAT报告基因表达以及阿斯匹林(环氧合酶1抑制剂)和NS-398(环氧合酶2抑制剂)对其的影响。结果重组人CD40配体使CAT报告基因表达下降;阿斯匹林和NS-398均可以增加CAT报告基因表达,并逆转重组人CD40配体诱导的CAT报告基因表达下降,NS-398的作用强于阿斯匹林(P<0.05)。结论重组人CD40配体对人Ⅰ型胶原α2(Ⅰ)链启动子活性的作用部分与环氧合酶通路有关。
Aim To evaluate the effects of cyclooxygenase （COX） inhibitor and recombinant human CD40 ligand （rhCD40L） on human a2 （Ⅰ） collagen promoter. Methods After amplification and identification, pCOLH22.4 and pCOLH21.6, a plasmid containing PCAT3-enhancer, were transfected into human vascular smooth muscle cell （VSMC） using Li-pofectamin transfection reagent. The expression of target gene and protein were measured by CAT enzyme linked immunosorbent assay （ELISA） kit. The effects of COX inhibitor and rhCD40L on CAT activity were evaluated after transfection. Results Compared with the control group, the CAT activities were inhibited by rhCD40L. NS-398 and aspirin could reverse the CAT activities inhibited by rhCD40L; the effect of NS-398 was stronger than that of aspirin. Condusion rhCD40L could inhibit the activities of human a2 （Ⅰ） collagen promoter by COX pathway partly.
出处
《中国动脉硬化杂志》
CAS
CSCD
2007年第2期81-84,共4页
Chinese Journal of Arteriosclerosis
基金
国家973子项目(2005CB523309)
