摘要
目的探讨严重脓毒症幼猪生化指标、肺部病理损伤与免疫机制间的相互关系,以及参附注射液的干预作用。方法将2~3月龄巴拿马香猪按随机数字表法分为假手术组(Sham组;静脉注射生理盐水)、脂多糖(LPS)致严重脓毒症模型组(LPS组;静脉注射LPS 1 mg/kg,0.5 mg·kg^-1·h^-1维持12 h)、参附注射液干预组(SF组;制模同时静脉注射参附注射液10 mL/kg,每日2次),每组5只。制模后48 h取血检测C-反应蛋白(CRP)、降钙素原(PCT)、氧合指数(PaO2/FiO2)、剩余碱(BE)、血乳酸(Lac)等生化指标;取外周血行流式细胞检测,分析中性粒细胞数目〔髓过氧化物酶阳性(MPO^+)〕及其激活亚群CD11b^+CD64^+、M1型巨噬细胞(CD80^+CD64^+)、CD4^+和CD8^+T细胞的变化;采用酶联免疫吸附试验(ELISA)检测血浆细胞因子水平;苏木素-伊红(HE)染色观察肺组织病理损伤;采用反转录-聚合酶链反应(RT-PCR)检测肺组织损伤相关分子及其受体、趋化因子、炎性因子、血管内皮相关分子、紧密连接蛋白的mRNA表达。结果与Sham组比较,LPS组CRP、PCT、Lac水平明显升高,PaO2/FiO2、BE明显下降;外周血CD80^+CD64^+、CD11b^+CD64^+、MPO^+、CD4^+和CD8^+T细胞数、肿瘤坏死因子-α(TNF-α)水平明显降低,白细胞介素-10(IL-10)、内皮细胞生长因子(VEGF)水平明显升高;肺组织高迁移率族蛋白B1(HMGB1)、晚期糖基化终末产物受体(RAGE)、促血管生成素2/1(Ang2/Ang1)的mRNA表达均明显升高,Toll样受体9(TLR9)、趋化因子(CXCL9、CXCL10)、TNF-α、IL-27、内皮细胞TEK酪氨酸激酶2(TIE2)、紧密连接蛋白血管内皮-钙黏蛋白(VE-CAD)和封闭蛋白(Occludin)的mRNA表达均明显下降;HE染色显示,肺泡腔炎性细胞浸润渗出、肺泡实变及肺泡间质层明显增厚、肺气肿。与LPS组比较,SF组Lac明显下降(mmol/L:4.2±1.0比6.3±1.1,P<0.05),BE、PaO2/FiO2明显升高〔BE(mmol/L):-6.4±2.6比-11.6±2.5,PaO2/FiO2(mmHg,1 mmHg=0.133 kPa):180±36比105±35,均P<0.0
Objective To investigate the relationship between the biochemical parameters,the pulmonary pathologic injury and the immune mechanism of severe sepsis in infant porcine,and the intervention effect of Shenfu injection.Methods Panamanian infant porcine(2-3 months old)were divided into sham operation group(Sham group;intravenous injection of normal saline),lipopolysaccharide(LPS)induced severe sepsis model group(LPS group;intravenous injection of LPS 1 mg/kg,and continuing at 0.5 mg·kg^-1·h^-1 for 12 hours),and Shenfu injection intervention group(SF group;intravenous injection of Shenfu injection 10 mL/kg at the same time of modeling,twice a day)according to the random number table method,with 5 in each group.Forty-eight hours after the challenge,the changes of C-reactive protein(CRP),procalcitonin(PCT),oxygenation index(PaO2/FiO2),base excess(BE),blood lactate(Lac)and other biochemical indexes were detected with blood sampling;the number of neutrophils[myeloperoxidase positive(MPO^+)]and their activated subsets CD11b^+CD64^+,M1 macrophages(CD80^+CD64^+),CD4^+and CD8^+T cells were analyzed in peripheral blood by flow cytometry.The levels of plasma cytokines were detected by enzyme linked immunosorbent assay(ELISA).The pathological damage of lung tissue was observed by hematoxylin-eosin(HE)staining.The mRNA expression of lung injury related molecules and their receptors,chemokines,cytokines,vascular endothelial related molecules and tight junction protein were detected by reverse transcription-polymerase chain reaction(RT-PCR).Results Compared with Sham group,the levels of CRP,PCT and Lac in LPS group significantly increased,and PaO2/FiO2 and BE significantly decreased.In the peripheral blood,CD80^+CD64^+macrophages,CD11b^+CD64^+and MPO+neutrophils,CD4^+,CD8^+T cells and tumor necrosis factor-α(TNF-α)significantly decreased,but interleukin-10(IL-10)and vascular endothelial cell growth factor(VEGF)significantly increased;the mRNA expressions of high mobility group protein B1(HMGB1),receptor for advanced glycation
作者
邢燕
程东良
史长松
Xing Yan;Cheng Dongliang;Shi Changsong(Department of PICU,Henan Provincial People's Hospital,People's Hospital of Zhengzhou University,Zhengzhou 450003,Henan,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2020年第6期696-701,共6页
Chinese Critical Care Medicine
基金
河南省医学科技攻关计划项目(2018020428)
河南省省直医疗机构医疗服务提升工程专科建设(培育)项目(2017-66)。
