热打击诱导血管内皮细胞组织因子早期表达分泌的初步研究

钱晶; 王凡凡; 万露露; 杨加乐; 施学智; 童华生; 周伟梁

doi:10.11855/j.issn.0577-7402.0408.2023.0807

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热打击诱导血管内皮细胞组织因子早期表达分泌的初步研究

基础研究 | 更新时间：2024-01-04

- 热打击诱导血管内皮细胞组织因子早期表达分泌的初步研究
- Primary study of heat stress inducing early expression and secretion of tissue factor in vascular endothelial cells
- 解放军医学杂志 2023年48卷第12期页码：1412-1419
- 作者机构：
  
  1.广州中医药大学研究生院，广东广州　510000
  2.南部战区总医院重症医学科，广东广州　510000
  5.南方医科大学第一临床医学院，广东广州　510000
  3.广州中医药大学深圳临床医学院，广东深圳　518000
  4.南部战区总医院急诊医学科，广东广州　510000
- 作者简介：
  
  钱晶，硕士研究生，主要从事重症中暑弥散性血管内凝血发病机制方面的研究
  童华生，E-mail：fimmuths@163.com
  周伟梁，E-mail：710833932@qq.com
- 基金信息：
  
  广东省自然科学基金(2022A1515010353);军事医学创新工程专项项目(18CXZ032)
- DOI：10.11855/j.issn.0577-7402.0408.2023.0807
  中图分类号：
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钱晶, 王凡凡, 万露露, 等. 热打击诱导血管内皮细胞组织因子早期表达分泌的初步研究[J]. 解放军医学杂志, 2023, 48(12): 1412-1419.

Qian Jing,Wang Fan-Fan,Wan Lu-Lu,et al.Primary study of heat stress inducing early expression and secretion of tissue factor in vascular endothelial cells[J].Medical Journal of Chinese People′s Liberation Army,2023,48(12):1412-1419.
钱晶, 王凡凡, 万露露, 等. 热打击诱导血管内皮细胞组织因子早期表达分泌的初步研究[J]. 解放军医学杂志, 2023, 48(12): 1412-1419. DOI： 10.11855/j.issn.0577-7402.0408.2023.0807.

Qian Jing,Wang Fan-Fan,Wan Lu-Lu,et al.Primary study of heat stress inducing early expression and secretion of tissue factor in vascular endothelial cells[J].Medical Journal of Chinese People′s Liberation Army,2023,48(12):1412-1419. DOI： 10.11855/j.issn.0577-7402.0408.2023.0807.

摘要

目的,2,探讨热打击诱导血管内皮细胞组织因子(TF)早期表达分泌的规律。,方法,2,将30只SPF级C57BL/6雄性小鼠随机分为对照组与热打击后复温0、3、6、9 h组(,n,=6)。热打击组小鼠暴露于动物孵箱热环境，核心温度达42.5 ℃时即为发生重症中暑。采用HE染色观察小鼠肝脏、肺脏及肾脏组织病理学变化；RT-qPCR检测小鼠组织,TF, mRNA表达水平；ELISA法检测小鼠血浆TF浓度。体外建立人脐静脉内皮细胞(HUVECs)热打击模型，分为对照组与热打击后复温0、3、6、9 h组。采用RT-qPCR、Western blotting及细胞免疫荧光化学技术分别检测HUVECs中,TF, mRNA及蛋白表达水平；采用ELISA法检测HUVECs培养上清中TF水平。,结果,2,对照组小鼠各组织未见明显病理损伤，热打击复温小鼠存在不同程度组织细胞结构性损伤及出血性、炎症性损伤。与对照组小鼠比较，热打击复温0、3 h组肾脏(1.719±0.018、1.241±0.178, vs. ,1.000±0.063)、复温3 h组肺脏(2.444±0.511, vs. ,1.000±0.106)及复温6 h组肝脏(7.312±0.618, vs. ,1.000±0.147)中,TF, mRNA表达量均明显升高(,P,<,0.05)。热打击小鼠复温0、3、6、9 h组血浆TF水平均较对照组小鼠升高[(132.426±17.920) pg/ml、(119.400±10.267) pg/ml、(107.374±13.495) pg/ml、(163.767±22.810) pg/ml, vs. ,(75.479±13.831) pg/ml，,P,<,0.01]。与对照组比较，热打击后复温6、9 h组HUVECs的,TF, mRNA表达水平明显升高(1.905±0.354、2.564±0.297, vs. ,1.000±0.097，,P,<,0.01)；热打击复温0、3、6、9 h组HUVECs培养上清TF水平均明显高于对照组[(36.309±4.101) pg/ml、(38.425±5.484) pg/ml、(41.655±4.380) pg/ml、(43.586±4.718) pg/ml, vs. ,(14.996±0.254) pg/ml，,P,<,0.01]。,结论,2,热打击血管内皮细胞中TF的早期表达分泌增加，可能是重症中暑循环TF的主要来源之一。

Abstract

Objective,2,To explore the pattern of early expression and secretion of tissue factor (TF) in vascular endothelial cells induced by heat stress.,Methods,2,Thirty SPF-rated C57BL/6 male mice were randomly divided into five groups: the control group and groups of indicated recovery time, including 0, 3, 6, and 9 h in room temperature after heat stress (,n,=6). Mice in the heat stress groups were exposed to an animal incubator to reach 42.5 ℃ for core body temperature for heat stroke. We analyzed the histopathological changes in the liver, lung, and kidney tissues with HE staining.,We measured the ,TF, mRNA in mice tissues by RT-qPCR and the plasma concentration of TF in mice with a commercial ELISA kit. Human umbilical vein endothelial cells (HUVECs) were placed in a culture incubator to build an ,in vitro, heat stress model. HUVECs were divided into five groups, including a control group and groups of indicated recovery time, including 0, 3, 6, and 9 h after heat stress. We quantified the expression of ,TF, mRNA and protein in HUVEC cells by RT-qPCR, Western blotting, and immunofluorescence and measured the secreted TF with a commercial ELISA kit.,Results,2,No significant pathological injury was observed in the tissues of the control group. Mice treated with heat stress had various degrees of structural injuries and hemorrhagic and inflammatory changes in multiple tissues. Compared to control group, the expression of ,TF, mRNA significantly increased in the kidney of heat stress-treated mice with 0 and 3 h recovery time (1.719±0.018, 1.241±0.178, vs. ,1.000±0.063), the lung with 3 h recovery time (2.444±0.511, vs. ,1.000±0.106) and the liver with 6 h recovery time (7.312±0.618, vs. ,1.000±0.147) (,P,<,0.05). The concentration of TF in plasma also sustainedly elevated in mice with 0, 3, 6, and 9 h recovery time after heat stress as compared to control group [(132.426±17.920) pg/ml, (119.400±10.267) pg/ml, (107.374±13.495) pg/ml, (163.767±22.810) pg/ml, vs. ,(75.479±13.831) pg/ml, respectively,P,<,0.01]. The expression levels of ,TF, mRNA were higher in heat stress HUVECs with 6 h and 9 h recovery time than the control cells (1.905±0.354, 2.564±0.297, vs. ,1.000±0.097,P,<,0.01). Secreted TF in the supernatant from HUVECs treated with heat stress and different recovery time also increased significantly [(36.309±4.101) pg/ml, (38.425±5.484) pg/ml, (41.655±4.380) pg/ml, (43.586±4.718) pg/ml, vs. ,(14.996±0.254) pg/ml,P,<,0.01].,Conclusion,2,Heat stress increased early expression and secretion of TF in vascular endothelial cells. Vascular endothelial cells may be a main source of circulating TF in heat stroke.

关键词

热打击血管内皮细胞组织因子表达分泌

Keywords

heat stressvascular endothelial cellstissue factorexpression and secretion

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