八肽胆囊收缩素对脂多糖诱导的大鼠肺泡巨噬细胞iNOS表达及NO生成的影响  

Effects of cholecystokinin-octapeptide (CCK-8) on iNOS expression and NO formation in rat alveolar macrophage after lipopolysaccharide challenge in vitro

作　　者：吴宗盛[1] 郑嘉奕[1] 姚咏明[2] 卢中秋[1] 

机构地区：[1]温州医学院附属第一医院急诊科,浙江温州325000 [2]解放军总医院第一附属医院全军烧伤研究所,北京100048

出　　处：《解放军医学杂志》2012年第6期578-580,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81071593);浙江省医学扶植重点建设学科计划(07-F04)~~

摘　　要：目的探讨八肽胆囊收缩素(CCK-8)对脂多糖(LPS)诱导的大鼠肺泡巨噬细胞(NR8383)诱导型一氧化氮合酶(iNOS)表达和一氧化氮(NO)生成的影响。方法体外培养大鼠巨噬细胞NR8383,分别加入LPS(终浓度1μg/ml)和不同浓度(10-10、10-8和10-6mol/L)的CCK-8进行干预,并设空白对照组(加入PBS)和丙谷胺干预组(丙谷胺终浓度2μg/ml,CCK-8浓度10-8mol/L,LPS浓度1μg/ml)。各组细胞培养24h后收集上清液,Griess法检测NO含量;取培养24h后的细胞,提取细胞总RNA,RT-PCR法检测iNOS基因表达情况;裂解培养24h后的细胞,离心取上清,生化法检测细胞内iNOS活性。结果 LPS可明显促进大鼠肺泡巨噬细胞iNOS的表达和NO的生成;预先加入不同浓度的CCK-8均可抑制LPS诱导的iNOS mRNA水平上升和活力增强,减少NO生成,且呈现剂量依赖性(P<0.05或P<0.01)。CCK受体拮抗剂丙谷胺可逆转CCK-8的抑制作用。结论 CCK-8可能通过降低LPS诱导的肺泡巨噬细胞iNOS表达和NO产生来实现其对内毒素休克动物的保护效应。Objective To investigate the effects of cholecystokinin-octapeptide（CCK-8） on LPS-induced inducible nitric oxide synthase（iNOS） expression and nitric oxide（NO） secretion of rat alveolar macrophage cell line（NR8383）.Methods NR8383 cells were cultured and pretreated with different concentrations of CCK-8（10-10,10-8 and 10-6mol/L） with or without 2μg/ml Proglumide（Pro） before LPS stimulation.The value of NO in the supernatant was measured by Griess reagent.The mRNA expression and activity of iNOS were determined by RT-PCR and biochemical detection,respectively.Results Compared with that of control group,LPS elevated the expression level of iNOS and the production of NO in rat alveolar macrophage cells significantly（P0.01）.CCK-8 obviously inhibited LPS-induced iNOS expression and activation of NR8383 cells,reduced NO generation in a concentration-dependent manner（P0.05 or P0.01）.Proglumide could reversed the inhibitory effect of CCK-8（P0.01）.Conclusions CCK-8 effectively inhibit the LPS-induced NO production via inhibiting expression and activity of iNOS in rat alveolar macrophages.This might be the underlying mechanism of CCK-8 in alleviating endotoxic shock（ES） in an animal model.

关 键 词：脂多糖类 巨噬细胞 肺泡 胆囊收缩素 一氧化氮合酶 

分 类 号：R563.9[医药卫生—呼吸系统]