八肽胆囊收缩素对脂多糖诱导ECV-304一氧化氮合酶表达的分子抑制作用  

Inhibitive effect of cholecystokinin octapeptide on iNOS expression induced by lipopolysaccharide in ECV-304

作　　者：高峰[1] 谷振勇[1] 平静[1] 刘霞[1] 赵丽[1] 徐锦荣[1] 倪志宇[1] 丛斌[1] 凌亦凌[1] 

机构地区：[1]河北医科大学法医系,石家庄050017

出　　处：《第三军医大学学报》2006年第19期1945-1948,共4页Journal of Third Military Medical University

基　　金：教育部科学技术研究资助重点项目(204016);河北省自然科学基金资助项目(301351)~~

摘　　要：目的探讨CCK-8抑制LPS诱导ECV-304 iNOS表达的分子机制。方法培养ECV-304,用溶剂、CCK-8、LPS及CCK受体拮抗剂丙谷胺分别或联合孵育细胞,用W estern b lot检测iNOS蛋白表达,用免疫细胞化学方法检测NF-κB P65蛋白表达与核移位,用RT-PCR检测CCK-AR和CCK-BR mRNA表达。结果①溶剂对照组ECV-304 iNOS蛋白呈低表达;LPS诱导iNOS表达明显上调,CCK-8剂量依赖性抑制LPS的作用,单独CCK-8对其无影响。②ECV-304存在CCK-AR和CCK-BR mRNA表达,其中CCK-BR mRNA表达稍强。LPS可诱导CCK-AR和CCK-BR mRNA表达明显增强。③溶剂对照组NF-κB P65蛋白主要分布于细胞质;LPS孵育1 h后细胞核中NF-κB P65表达明显增加,CCK-8剂量依赖性抑制LPS的作用;单独CCK-8对其无影响。④丙谷胺可翻转CCK-8的抑制作用。结论ECV-304存在CCK-AR和CCK-BR mRNA表达;CCK受体和NF-κB参与介导CCK-8抑制LPS诱导ECV-304 iNOS蛋白表达上调。Objective To elucidate the molecular mechanism of cholecystokinin oetapeptide （CCK-8） inhibiting of iNOS expression in ECV-304 induced by LPS. Methods ECV-304 cells, were stimulated with LPS in the presence or absence of CCK-8 （10^-9 - 10^-7mol/L） or/and CCK receptor antagonist proglumide. The iNOS protein level in the cytoplasm at 16 h after stimulation was detected with Western blotting. The expressions of CCK-AR and CCK-BR mRNA were assayed with RT-PCR. The level of NF-κB P65 protein was detected in cytoplasm and nuelus at 1 h after stimulation with immunoeytoehemieal technique. Results LPS led to the up-regulation of iNOS protein expression in ECV-304 that was obviously inhibited by CCK-8 in a dose-dependent manner. CCK-8 alone did not affect iNOS protein expression. CCK-AR mRNA expression was higher than CCK-BR mRNA expression in ECV-304. The expression of CCK-AR and CCK-BR mRNA could be upregulated obviously by LPS. LPS increased NF-κB P65 protein expression in the nuelus of ECV-304. CCK-8 obviously inhibited LPS-indueed changes of NF-κB P65 protein in a dose-dependent manner. The inhibitive effects of CCK-8 on iNOS protein expression and NF-κB P65 were attenuated by proglumide. Conclusion There were CCK-AR and CCK-BR mRNA expressions in ECV-304. CCK-8 inhibited LPS-indueed up-regulation of iNOS protein expression by acting CCK receptors and NF-κB in ECV-304.

关 键 词：缩胆囊素 脂多糖类 诱生型一氧化氮合酶 核转录因子-κB CCK受体 

分 类 号：R345[医药卫生—基础医学] R394.6