| 王 倩,徐玲文,揭凤英,孙 雯,魏文娇,谌 芳,董 芳.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[J].,2024,(11):2032-2037 |
| 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 |
| 投稿时间：2024-01-09 修订日期：2024-01-31 |
| DOI：10.13241/j.cnki.pmb.2024.11.006 |
| 中文关键词: GM-CSF BMSCs 脓毒症 小鼠 血管内皮损伤 凝血功能 炎症因子 |
| 英文关键词: GM-CSF BMSCs Sepsis Mice Vascular endothelial injury Coagulation function Inflammatory factors |
| 基金项目:武汉市卫健委西医青年项目（WX20C27）；湖北省科技厅计划项目（2019CFB789） |
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| 中文摘要: |
| 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 mRNA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。 |
| 英文摘要: |
| ABSTRACT 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 significantly increased, PT, APTT time, serum IL-6, TNF-α, IL-1 level, apoptosis rate, TF, TFPI, VCAM1 and PAR1 protein expression in vascular endothelial tissue were significantly decreased (P<0.05). The detection indexes in GM-CSF-BMSCs group were better than those in BMSCs group (P<0.05). Conclusion: GM-CSF transfected BMSCs can reduce the inflammatory response and vascular endothelial cell injury in septic mice, and improve coagulation function. |
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