高峰 谷振勇 平静 王杏云 刘霞 徐锦荣 赵丽 闫玉仙 马丽琴 丛斌 凌亦凌

    【摘要】 目的 探讨八肽缩胆囊素(CCK8)调节脂多糖(LPS)诱导血管内皮细胞核转录因子κB(NFκB)表达的受体机制。 方法 培 养人脐静脉内皮细胞株ECV304；用溶剂(生理盐水)、LPS、CCK8、CCK受体(CCKR)非特异性拮抗剂丙谷胺、CCKA受体(CCKAR)特异性拮抗剂CR1409、CCKB受体(CCKBR)特异性拮抗剂CR2945分别或联合刺激ECV304细胞1 h。用蛋白质免疫印迹法(Western blot)检测NFκB p65蛋白表达；用免疫细胞化学技术检测NFκB p65蛋白核移位。结果 与溶剂对照组比较，LPS可诱导ECV304细胞NFκB p65蛋白核移位，且其表达明显上调；CCK8可呈剂量依赖性地抑制LPS诱导的核移位及表达上调；CCK受体拮抗剂可翻转CCK8的上述抑制效应，其中CR1409、CR2945、丙谷胺作用依次增强。结论 CCKAR和CCKBR参与介导了CCK8对LPS诱导ECV304细胞NFκB表达的抑制作用，其中CCKBR的作用比CCKAR稍强。

    【关键词】 八肽缩胆囊素；内毒素；缩胆囊素受体；核转录因子κB

    Receptor mechanisms underlying the modulation of lipopolysaccharideinduced nuclear factorκB expression in vascular endothelial cells by cholecystokinin octapeptide GAO Feng, GU Zhenyong, PING Jing, WANG Xingyun, LIU Xia, XU Jinrong, ZHAO Li, YAN Yuxian, MA Liqin, CONG Bin, LING Yiling. Department of Forensic Medicine and Pathophysiology, Shijiazhuang 050017, Hebei, China (GAO Feng works at Department of Forensic Medicine, Public Security Bureau of Hebei Province, Shijiazhuang, Hebei, China)

    【Abstract】Objective To elucidate the receptor mechanisms underlying the modulation of lipopolysaccharide (LPS)induced nuclear factorκB (NFκB) expression in human umbilical vein endothelial cell line ECV304 cells by cholecystokinin octapeptide (CCK8). Methods Human umbilical vein endothelial cell line ECV304 cells were stimulated with vehicle, LPS, CCK8 (10-910-7 mol/L), CCK receptor nonspecific antagonist proglumide, CCKA receptor (CCKAR) specific antagonist CR1409 or CCKB receptor (CCKBR) specific antagonist CR2945 singularly or in combination. The NFκB p65 protein level was determined by Western blot and immunocytochemistry technique. Results LPS resulted in an increase in the upregulatory expression and nuclear translocation of NFκB p65 protein in ECV304 compared with vehicle stimulation. CCK8 obviously inhibited LPSinduced the changes in NFκB p65 protein in a dosedependent manner. The inhibitory effects of CCK8 on NFκB p65 protein expression were attenuated by proglumide>CR2945>CR1409. Conclusion CCKAR and CCKBR are involved in the mediation of CCK8 inhibitive regulation for LPSinduced NFκB protein expression in ECV304 cells, whereas the effect of CCKBR are more than that of CCKAR. ......