GM-CSF基因转染的BMSCs对脓毒症小鼠血管内皮损伤、凝血功能及炎症因子的影响  

Effects of GM-CSF Gene Transfected BMSCs on Vascular Endothelial Injury,Coagulation Function and Inflammatory Factors in Septic Mice

作　　者：王倩 徐玲文 揭凤英 孙雯 魏文娇 谌芳 董芳 WANG Qian;XU Ling-wen;JIE Feng-ying;SUN Wen;WEIWen-jiao;CHEN Fang;DONG Fang(Department of Intensive Care Medicine,Wuhan Third Hospital,Wuhan,Hubei,434200,China)

机构地区：[1]武汉市第三医院重症医学科,湖北武汉434200

出　　处：《现代生物医学进展》2024年第11期2032-2037,共6页Progress in Modern Biomedicine

基　　金：武汉市卫健委西医青年项目(WX20C27);湖北省科技厅计划项目(2019CFB789)。

摘　　要：目的:探讨粒细胞-巨噬细胞集落刺激因子(GM-CSF)基因转染的骨髓间充质干细胞(BMSCs)对脓毒症小鼠血管内皮损伤、凝血功能及炎症因子的影响。方法:将60只SD小鼠分为Sham组、模型(Model)组、BMSCs组、GM-CSF-BMSCs组,每组15只,采用盲肠结扎穿孔法制备小鼠脓毒症模型。实时荧光定量聚合酶链式反应(qRT-PCR)检测细胞GM-CSF基因表达;记录24 h小鼠尾出血时间;检测血浆凝血酶原时间(PT)、活化部分凝血酶原时间(APTT)、凝血酶时间(TT)、纤维蛋白原(Fib)水平;ELISA法检测血清白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、IL-1、IL-10水平;苏木精-伊红(HE)染色观察血管组织形态;TUNEL法检测血管内皮细胞凋亡;Western blot检测血管组织因子(TF)、组织因子途径抑制物(TFPI)、抗凝血酶(ATⅢ)、血管细胞粘附因子1(VCAM1)、蛋白酶激活受体(PAR1)蛋白表达。结果:与CK组BMSCs比较,Transfection组BMSCs细胞GM-CSF m RNA表达显著升高(P<0.05)。与Sham组比较,Model组小鼠血管形态结构有明显的损伤,小鼠尾出血时间、Fib含量、血清IL-10水平和血管内皮组织ATⅢ蛋白表达显著降低,PT、APTT、TT时间、血清IL-6、TNF-α、IL-1水平、凋亡率、血管内皮组织TF、TFPI、VCAM1、PAR1蛋白表达显著升高(P<0.05);与Model组比较,BMSCs组和GM-CSF-BMSCs组小鼠血管形态结构损伤明显减轻,小鼠尾出血时间、Fib含量、血清IL-10水平和血管内皮组织ATⅢ蛋白表达显著升高,PT、APTT时间、血清IL-6、TNF-α、IL-1水平、凋亡率、血管内皮组织TF、TFPI、VCAM1、PAR1蛋白表达显著降低(P<0.05);GM-CSF-BMSCs组各项检测指标优于BMSCs组(P<0.05)。结论:GM-CSF转染的BMSCs可减轻脓毒症小鼠炎症反应和血管内皮细胞损伤,改善凝血功能。Objective:To investigate the effects of granulocyte-macrophage colony-stimulating factor(GM-CSF)gene transfected bone marrow mesenchymal stem cells(BMSCs)on vascular endothelial injury,coagulation function and inflammatory factors in septic mice.Methods:60 SD mice were divided into Sham group,model(Model)group,BMSCs group and GM-CSF-BMSCs group,with 15 mice in each group,sepsis model of mice was prepared by cecal ligation and puncture.The expression of GM-CSF gene was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The tail bleeding time of 24h mice was recorded.The levels of plasma prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT)and fibrinogen(Fib)were detected.The levels of serum interleukin(IL)-6,tumor necrosis factor(TNF)-α,IL-1 and IL-10 were detected by ELISA.The morphology of vascular tissue was observed by hematoxylin-eosin(HE)staining.Apoptosis of vascular endothelial cells was detected by TUNEL method.The expression of vascular tissue factor(TF),tissue factor pathway inhibitor(TFPI),antithrombin(ATIII),vascular cell adhesion molecule 1(VCAM1)and protease-activated receptor 1(PAR1)were detected by Western blot.Results:Compared with BMSCs in CK group,the expression of GM-CSF mRNA in BMSCs in Transfection group was significantly increased(P<0.05).Compared with Sham group,the vascular morphology of the mice in Model group was significantly damaged,the tail bleeding time,Fib content,serum IL-10 level and ATIII protein expression in vascular endothelial tissue of the mice in Model group were significantly decreased,PT,APTT,TT time,serum IL-6,TNF-α,IL-1 levels,apoptosis rate,TF,TFPI,VCAM1,PAR1 protein expression in vascular endothelial tissue were significantly increased(P<0.05).Compared with Model group,the damage of vascular morphology and structure in BMSCs group and GM-CSF-BMSCs group was significantly reduced,the tail bleeding time,Fib content,serum IL-10 level and ATIII protein expression in vascular endothelial tissue were significan

关 键 词：GM-CSF BMSCS 脓毒症 小鼠 血管内皮损伤 凝血功能 炎症因子 

分 类 号：R-33[医药卫生] R631.2