胍丁胺对创伤小鼠过度炎症反应及脾细胞增殖的影响  

Effects of agmatine on excessive inflammatory reaction and proliferation of splenic cells in mice with trauma

作　　者：刘政[1] 侯凤艳[1] 靳贺 肖雅[1] 范霞[1] 杨雪[1] 严军[1] 梁华平[1] 

机构地区：[1]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,重庆400042

出　　处：《中华危重病急救医学》2015年第2期110-114,共5页Chinese Critical Care Medicine

基　　金：基金项目：国家重点基础研究发展计划（973）项目（2012CB518102）

摘　　要：目的 探讨胍丁胺对创伤小鼠炎症反应及脾细胞免疫功能的影响。方法 将48只成年雄性C57BId6小鼠按随机数字表法分为对照组、模型组（双后肢骨折＋失血35％）和胍丁胺组（创伤／失血＋胍丁胺200mg／kg），每组16只。各组分别于制模后3h、24h处死8只小鼠，采集血、脾脏及肝脏。采用酶联免疫吸附试验（ELISA）检测血清和肝组织中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-6、IL-1β）含量，用全自动生化分析仪测定血清天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）及乳酸脱氢酶（LOI~）水平，采用四甲基偶氮唑盐比色法（MTT）检测刀豆蛋白A（ConA）刺激的脾细胞增殖反应，用ELISA法检测ConA刺激脾细胞分泌γ-干扰素（IFN-γ）与IL-2的能力。结果 与对照组比较，模型组创伤后3h血清TNF—α、IL-6、IL-1β水平均显著升高[TNF—α（ng／L）：145．38±31．50比23．06±11．14；IL-6（ng/L）：496．94±50．76比47．13±17．47；IL-18（ng／L）：321．31±43．02比29．25±16．24；均P〈O．01]，胍丁胺治疗能明显抑制创伤诱导的血清炎症因子TNF-α（ng／L：111．56±25．47比145．38±31．50）、IL-6（ng／L：412．56±44．33比496．94±50．76）、IL-1β（ng/L：273．38±45．25比321．31±43．02）的升高（P〈0．05或P〈0．01）；创伤后24h，模型组和胍丁胺组血清炎症因子均恢复至对照组水平。创伤后3h，各组肝组织TNF—α、IL-6水平无明显差异；创伤后24h，模型组肝组织TNF-β、IL-6水平均较对照组显著升高[TNF-α（ng／mg）：32．93±4．90比26．58±2．33；IL-6（ng／mg）：11．20±1．66比8．38±0．89；均P〈0．01]，胍丁胺治疗能抑制创伤引起的肝组织TNF—α（ng／mg：28．92±3．16比32．93±4．90）、IL-6（ng／mg：9．03±1．28比11．20±1．66）水平升高（P〈0．05和P〈O．01）。创伤后24h，模型组和胍丁胺组血清AST、ALT、LDH均较Objective To observe protective effects of agmatine （AGM） on inflammatory response and spleen immune function in mice with trauma. Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups （n = 16 each）, including control group, model group （bilateral femoral fracture and removal of 35% of the total blood volume ）, and AGM group （trauma/hemorrhage ＆ AGM 200 mg/kg ）. Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α （TNF-α ）, intedeukins （IL-6, IL-1β ） in serum and liver tissue were determined with enzyme linked immunosorbent assay （ELISA）. Serum aspartate transaminase （AST）, alanine aminotransferase （ALT） and lactic dehydrogenase （LDH） were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A （ConA） was evaluated with methyl thiazolyl tetrazolium colourimetry （MTT）. γ-interferon （IFN-γ, ） and IL-2 releases were determined with ELISA. Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF- α, IL-6, and IL-1β in model group were significantly elevated [TNF-α （ng/L）： 145.38± 31.50 vs. 23.06 ±11.14, IL-6 （ng/L）： 496.94 ± 50.76 vs. 47.13± 17.47, IL-1β （ng/L）： 321.31 ±43.02 vs. 29.25 ± 16.24, all P 〈 0.01 ]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediator~ induced by trauma/hemorrhage, such as TNF-α （ng/L： 111.56±25.47 vs. 145.38±31.50）, IL-6 （ng/L： 412.56±44.33 vs. 496.94±50.76）, IL-1β （ng/L： 273.38±45.25 vs. 321.31±43.02, P 〈 0.05 or P 〈 0.01 ）. Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups.

关 键 词：胍丁胺 创伤 炎症反应 免疫调节 

分 类 号：R965[医药卫生—药理学]