多巴胺对脂多糖诱导小鼠腹腔巨噬细胞炎症反应的影响  

Effect of dopamine on lipopolysaccharide-induced inflammatory response in mouse peritoneal macrophages

作　　者：李琳[1] 李磊[2] 王慧丽[4] 谭燕[2] 吴晓凤[2] 刘锐[3] 温秀杰[3] 刘畅[1] 唐明[1] 

机构地区：[1]重庆医科大学附属第一医院口腔科,400016 [2]第三军医大学大坪医院野战外科研究所第一研究室、创伤、烧伤与复合伤国家重点实验室 [3]第三军医大学大坪医院口腔科 [4]西安外事学院医学院护理系

出　　处：《中华创伤杂志》2016年第6期552-557,共6页Chinese Journal of Trauma

基　　金：国家重点基础研究发展计划，重庆市基础科学与前沿技术研究项目(cstc2014jcyjA1217) National Key Basic Research Program of China，Chongqing Research Program Basic Science and Advanced Technology

摘　　要：目的探讨多巴胺对脂多糖（LPS）诱导小鼠腹腔巨噬细胞（MPMs）炎症反应的影响。方法选择28只C57雄性Toll样受体4（TLR4）基因野生型（TLR4＋/＋）小鼠和6只雄性TLR4基因缺失型（TLR4-/-）小鼠，随机进行腹腔注射巯基乙酸盐肉汤，获取原代MPMs。（1）TLR4＋/＋MPMs。分为对照组、LPS组、多巴胺预处理组、多巴胺D1类受体拮抗剂组（D1拮抗剂组）、多巴胺D2类受体拮抗剂组（D2拮抗剂组），其中后两组分别为多巴胺D1类受体拮抗剂和D2类受体拮抗剂处理MPMs30min后再用多巴胺＋LPS处理；（2）TLR4＋/＋和TLR4-MPMs。分为LPS组和多巴胺预处理组。LPS组用1μg／mlLPS刺激MPMs6h，多巴胺预处理组用10^-多巴胺作用MPMs2h后再用1μg/mlLPS刺激6h。Westernblot检测各组细胞TLR4、白细胞介素-1β前体（pro-IL-1β）的表达，酶联免疫吸附（ELISA）法检测细胞上清中肿瘤坏死因子-α（TNF-α）的表达。结果（1）TLR4＋/＋ MPMsTLR4、pro-IL-1α和TNF-α的表达情况：对照组为0．56±0．07、0．65±0．11和（1770．6±448．8）pg／ml，LPS组为1．12±0．15、1．24±0．20和（15569．5±822．7）pg／ml，多巴胺预处理组为0．28±0．11、0．22-4-0．08和（7800．7±862．6）pg／ml，DI拮抗剂组为0．25±0．12、0．18±0．09和（7065．0±1016．8）pg／ml，D2拮抗剂组为0．80±0．09、0．44±0．08和（14299．6±1430．9）pg／ml。其中LPS组均高于对照组和多巴胺预处理组（P均〈0．05），D2拮抗剂组均高于多巴胺预处理组（P〈0．05），而D1拮抗剂组与多巴胺预处理组差异无统计学意义（P〉0．05）。（2）pro—IL-1β和TNF—α的表达情况：LPS组TLR4＋/＋MPMs为0．94±0．17和（15109．0±1903．4）pg／ml，TLR4-/-MPMs为0．08±0．04和（5063．6±512．8）pg／ml；多巴胺预处理组TLR4＋/＋MPMs为0．45±0．12和（6383．9±1287．3）pg／ml，TLR4-/-MPMs为0．05±0.02和（4863．0±824．7）pg�Objective To investigate the effect of dopamine on lipopolysaccharide （LPS）- induced inflammatory response in mouse peritoneal macrophages （MPMs）. Methods MPMs wereisolated after injection of thioglycolate broth into the peritoneal cavity. MPMs from wild type C57 mice were distributed into control group, LPS group, dopamine pretreatment group, dopamine Dl-like and and D2-1ike receptor antagonist groups （D1 and D2 antagonist groups）. In the latter two groups, MPMs were pretreated with Dl-like and D2-1ike receptor antagonist respectively for 30 min, and then stimulated with dopamine and LPS. MPMs from wild type （TLR4＋/＋ ） and TLR4 knock-out （TLR4-/- ） mice were only divided into LPS group and dopamine pretreatment group. In LPS group, MPMs were stimulated with 1 μg/ml LPS for 6 h. In dopamine pretreatment group, MPMs were pretreated with 10-4mol/L dopamine for 2 h, and then stimulated with 1 μg/ml LPS for 6 h. Expressions of TLR4 and pro-IL-1βwere detected by Western blot and tumor necrosis factor-α （TNF-α） in cell culture supernatant by ELISA method. Results （ 1 ） Expressions of TLR4, pro-IL-1 β and TNF-α in control group were （0.56 ± 0.07 ）, （0.65± 0.11 ） and （ 1,770.6 ± 448.8 ） pg/ml ; in LPS group were （ 1.12 ± 0. 15 ）, （ 1.24 ± 0.20） and （ 15, 569.5 ±822.7） pg/ml ; in dopamine pretreatment group were （0.28 ± 0.11 ）, （0.22 ± 0.08 ） and （ 7, 800.7 ±862.6）pg/ml; in D1 antagonist group were （0.25 ±0.12）, （0.18 ±0.09） and （7,065.0± 1016.8）pg/ml; in D2 antagonist group were （0.80 ±0.09）, （0.44 ±0.08） and （14,299.6 ± 1430. 9）pg/ml. The three indicators in LPS group were increased compared to control group and dopamine pretreatment group （ P 〈 0. 05 ）, and in D2 antagonist group were increased compared to dopamine pretreatment group （ P 〈 0. 05 ）. There were no obvious differences between D1 antagonist group and dopamine pretreatment group（ P 〉 0.05 ）. （ 2 ） After LPS stimulation, exp

关 键 词：多巴胺 炎症 巨噬细胞  腹膜 多巴胺受体 

分 类 号：R392[医药卫生—免疫学]