CC趋化因子ligand-5促进在肝脏缺血再灌注损伤早期巨噬细胞浸润的机制  

CC chemokine ligand-5 promotes macrophage infiltration in early stage of hepatic ischemia-reperfusion injury

作　　者：张增光[1] 孙贺元[2] 王凤飚[1] Zhang Zengguang;Sun Heyuan;Wang Fengbiao(Department of Hepatobiliary Surgery,Tianjin 4th Central Hospital,Tianjin 300143,China;Heavy Disease Medicine Division,Tianjin 4th Central Hospital,Tianjin 300143,China)

机构地区：[1]天津市第四中心医院肝胆胃肠外科,300143 [2]天津市第四中心医院重症医学科,300143

出　　处：《中华实验外科杂志》2020年第1期48-51,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨CC趋化因子ligand-5(CCL5)促进在肝脏缺血再灌注损伤早期巨噬细胞浸润的机制。方法使用成年雄性C57BL/6野生型和CCL5缺陷小鼠进行实验,随机分为4组:假手术组(假手术组小鼠经历缺血灌注模型方案,但没有进行血管闭塞,n=10);缺血再灌注模型组(用异氟烷麻醉小鼠,进行中线剖腹手术,并将一个无创伤夹子放置在静脉门、肝动脉和胆管上,以中断左侧外侧叶和中叶的血液供应,在部分肝脏缺血60 min后,移除夹子以开始再灌注,n=10);CCL5拮抗剂+模型组[在即将开始再灌注之前,CCL5拮抗剂+模型组接受单次推注静脉注射趋化因子受体5(CCR5)受体拮抗剂马拉维诺,n=10];CCL5缺陷组(CCL5缺陷小鼠,n=10);通过收集小鼠血液用VTROS DT60Ⅱ化学系统检测谷丙转氨酶(ALT)含量及髓过氧化物酶(MPO)活性;通过实时定量反转录聚合酶链反应(RT-qPCR)检测ALT mRNA及MPO mRNA的表达;通过流式细胞术检测巨噬细胞的数量;通过组织学和免疫组织化学检测巨噬细胞对肺部的浸润情况;通过蛋白质免疫印迹检相关炎性因子的蛋白表达。通过单因素方差分析组间的统计学差异。结果与假手术组比较缺血再灌注模型组中ALT含量[(521.36±20.65)比(4126.55±326.68),F=16.237,P<0.05]及MPO[(1.78±0.54)比(7.66±2.34),F=14.211,P<0.05]活性升高,CCL5拮抗剂+模型组较缺血再灌注模型组降低(P<0.05),差异有统计学意义。在前1~3 h缺血再灌注模型组与CCL5缺陷组中CD45+数量比较差异无统计学意义(P>0.05),在第24小时缺血再灌注模型组较CCL5缺陷组CD45+的数量升高[(2.91±0.23)比(1.65±0.17),F=16.311,P<0.05]。再灌注后1、2、8和24h中,发现缺血再灌注模型组CD68+细胞数量较假手术组高[(94.62±8.55)比(23.68±3.11),F=16.352,P<0.05],差异有统计学意义,而在缺血再灌注早期CCL5缺陷组中较缺血再灌注模型组降低[(76.38±6.77)比(94.62±8.55),F=16.352,P<0.05],差异有统计学意义。与与假手�Objective To study the mechanism of CC chemokine ligand-5 promoting macrophage infiltration in early stage of hepatic ischemia-reperfusion injury.Methods The adult male C57BL/6 wild type and CC chemokine ligand-5(CCL5)deficient mice were used for the experiment.The mice were kept in captivity at(23±2)℃,and were exposed to light and dark cycles for 12 hours to obtain food and water freely.The mice were randomly divided into 4 groups.Sham operation group(sham operation group mice experienced ischemia-reperfusion model scheme,but no vascular occlusion,n=10).In the ischemia-reperfusion model group(mice were anesthetized with isoflurane and underwent midline laparotomy,and a noninvasive clamp was placed on the portal,hepatic artery and bile duct to interrupt the blood supply of the left lateral lobe and middle lobe.After 60 minutes of partial liver ischemia,the clamp was removed to start reperfusion,n=10).CCL5 antagonists+model group(before the start of reperfusion,the CCL5 antagonists+model group received a single intravenous injection of the CCR5 receptor antagonist maladono,n=10);CCL5 defective group(CCL5 defective mice,n=10).Alanine aminotransferase(ALT)content and myeloperoxidase(MPO)activity were measured by vtros dt60Ⅱchemical system.The expression of ALT mRNA and MPO mRNA was detected by eal-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).The number of macrophages was detected by flow cytometry.The infiltration of macrophages into the lung was detected by histology and immunohistochemistry.The protein expression of related inflammatory factors was detected by Western blotting.Results Compared with the sham operation group,ALT content[(521.36±20.65)vs.(4126.55±326.68),F=16.237,P<0.05]and MPO[(1.78±0.54)vs.(7.66±2.34),F=14.211,P<0.05]activity in the ischemia-reperfusion model group increased.CCL5 antagonist+model group was lower than ischemia-reperfusion model group(P<0.05).There was no difference in the number of CD45+between the ischemia-reperfusion model group and CCL5 defi

关 键 词：肝脏缺血再灌注 巨噬细胞 CC趋化因子ligand-5 炎性反应 拮抗剂 

分 类 号：R575[医药卫生—消化系统]