热打击及中暑小鼠血清对小鼠肺微血管内皮细胞黏附单核细胞能力的影响  

Effects of heat stress and the serum of mice suffering from heat stroke on adherent ability of monocytes to pulmonary microvascular endothelial cells in vitro

作　　者：刘亚楠[1,2] 徐秋林[1] 郭晓华[2] 周耿标 王郑莲 陆杰富 邱俊明[1] 苏磊[1] 

机构地区：[1]广州军区广州总医院重症医学科,广东广州510010 [2]南方医科大学研究生学院,广东广州510515 [3]广东中医药大学,广东广州510515

出　　处：《感染．炎症．修复》2014年第3期131-135,F0002,共6页Infection Inflammation Repair

基　　金：广东省自然科学基金博士启动项目(S2013040015661);广东省医学科研基金资助项目(B2010176);军队"十二五"医学科研基金重点项目(BWS12J018)

摘　　要：目的:研究热打击及10%中暑小鼠血清刺激肺微血管内皮细胞(PMVECs)对PMVECs表达血管内皮黏附分子(VCAM-1)和细胞间黏附分子(ICAM-1)水平及其黏附单核细胞能力的影响。方法:根据二次磁珠分选法分离乳鼠PMVECs,光镜下观察细胞形态并对其血管内皮细胞钙黏蛋白(VE-Cadherin)及Ⅷ因子进行染色鉴定。将PMVECs随机分为3组,分别进行37℃正常培养(对照组)、给予42℃2 h热打击或10%中暑小鼠血清刺激,24 h后分别提取总RNA,荧光定量PCR及Weston blot检测VCAM-1及ICAM-1mRNA和蛋白的表达水平,荧光显微镜观察其对人单核细胞(THP-1细胞)的黏附能力。结果:所分离PMVECs纯度较高,光镜下细胞呈特征性鹅卵石形状排列,VE-Cadherin及Ⅷ因子均可染色。与对照组相比,给予42℃2 h的热打击24 h后,VCAM-1及ICAM-1的mRNA及蛋白水平表达降低(P<0.05),而给予10%中暑小鼠血清刺激24 h后VCAM-1及ICAM-1的mRNA及蛋白水平表达增加(P<0.01)。与对照组相比,热打击组黏附THP-1细胞的细胞数无明显变化,中暑小鼠血清刺激组THP-1细胞数明显增多,显示PMVECs对单核细胞的黏附能力增强。结论:单独热打击并未使小鼠PMVECs黏附分子表达增加,而给予中暑小鼠血清刺激后其黏附分子表达显著上调,从而使单核细胞易于附壁,导致或加重了中暑相关的急性肺损伤。Objective：To investigate the effect of heat stress and the serum of mice suffering from heat stroke on capacity of pulmonary microvascular endothelial cells（PMVECs） to express vascular cell adhesion molecule-1（VCAM-1） and intercellular adhesion molecule-1（ICAM-1）,and adherent capacity of PMVECs to monocytes in vitro.Methods：PMVECs were isolated from neonatal mice by the method of secondary magnetic bead sorting.The morphology of PMVECs was observed under light microscope and identified by VE-Cadherin and Ⅷ factor staining.PMVECs were randomly divided into 3 groups,including cells undergoing normal culture（37℃,as normal control）,42℃ heat stress for 2 hours,and co-culture with serum of mice with heat stroke.Twenty-four hours later,total RNA was extracted,the quantitative PCR and Western blotting were used for the determination of mRNA and protein expressions of VCAM-1 and ICAM-1.The adherent capacity of PMVECs to monocytes was observed with fluorescence microscopy.Results：The PMVEC displayed typical ＂cobblestone＂ morphology under light microscopy,and they were positive with both VE-cadherin staining at cell-cell junctions and Ⅷ factor,thus they were identified as endothelial cells.Compared to normal control group,the VCAM-1 and ICAM-1 mRNA and protein levels were decreased in heat stress group（P〈0.05）,but increased after stimulation of 10%serum of mice with heat stroke（P〈0.01）.Compared to the normal control group,the number of adherent THP-1 cells was similar in heat stress group,but the number of THP-1 cells was significantly increased in serum stimulation group,showing that the adherent capacity of PMVECs to monocyte was enhanced.Conclusions：Heat stress did not upregulate the expression of endothelial cell adhesive molecules of PMVECs.But stimulation of serum of mice with heat stroke significantly up-regulate the expression of endothelial cell adhesion molecules,facilitating adhesion of monocytes,which produce or aggravate acute lung injury in heat stroke.

关 键 词：热打击 肺微血管内皮细胞 单核细胞 黏附分子 中暑 

分 类 号：R735.1[医药卫生—肿瘤]