首页|期刊导航|军事医学|失血大鼠肠道多节段病理学评价及巨噬细胞功能研究

失血大鼠肠道多节段病理学评价及巨噬细胞功能研究OACSTPCD

Pathological evaluation of multiple intestinal segments and the macro-phage function study in hemorrhage rats

中文摘要英文摘要

目的 研究重度失血大鼠模型中各肠道节段的病理损伤和炎症情况随时间变化的特点,探索肠道巨噬细胞极化对肠道炎症病理生理的可能影响.方法 将雄性Wistar大鼠1∶1随机分为假手术组和失血组,失血组股动静脉插管并在25~30 min快速失去大鼠总血量的40%,假手术组仅插管未失血,然后将上述两组大鼠分别1∶1∶1∶1∶1随机分为5组,分别以放血完成后0、3、6、12和24 h作为实验终点处死,快速分离全肠并在十二指肠、空肠、回盲部、结肠和直肠相同位置剪取肠段进行病理组织学评分、酶联免疫吸附测定相关炎症因子以及多色免疫组织化学染色测定巨噬细胞表面标志物.结果 (1)与假手术组相比,结肠3、6h时的组织学评分差异无统计学意义,直肠除24 h以外各时间点评分差异无统计学意义,其余各肠段0h后各时间点的评分与假手术组相比,差异均具有统计学意义;回盲部、结肠组织损伤程度随失血时间的延长逐渐加重;同一失血时间不同肠段相比,3 h时十二指肠、空肠、回盲部的损害程度显著重于结肠、直肠,6 h时十二指肠、空肠、回盲部的损害程度显著重于直肠,12h时十二指肠、空肠、回盲部与结肠的损害程度显著重于直肠,24 h时回盲部的损害程度与直肠相比差异具有统计学意义.(2)与假手术组相比,重度失血大鼠十二指肠段12 h后肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)表达显著上调;空肠段IL-1β在3 h后表达显著上调,TNF-α则在6 h表达显著上调;回盲部、结肠、直肠段重度失血在3 h时即引起IL-1β、TNF-α表达显著增加.(3)重度失血后大鼠3 h时空肠、回盲部的巨噬细胞极化细胞显著增加,M1型比例更高、增加更多,6 h后极化巨噬细胞比例逐渐降低至失血前水平,6h时空肠的M2型巨噬细胞占比下降更多,回盲部M1型巨噬细胞占比下降更多.结肠3h后M1型巨噬细胞比例下降,3 h时M2型巨噬细胞有增加趋势.重度失血3h时十二指肠、直肠M2型极化巨噬细胞占比增加,6h后有所减少.结论 不同肠道节段在失血发生后病理损伤程度有差异,并且存在一定的时间变化规律,与巨噬细胞炎症水平也有一定相关性.

Objective To investigate the pathological damage to and inflammation of different intestinal segments in a rat model of severe hemorrhage,and to explore the effect of polarization of intestinal macrophage on the pathophysiology of intestinal inflammation.Methods Male Wistar rats were randomly divided into two groups:the sham operation group and hemorrhage group.In the hemorrhage group,40%of the total blood volume was lost in 25-30 minutes,while in the sham operation group,only the femoral artery and vein were intubated without bleeding.The rats were killed at 0,3,6,12 and 24 hours.The entire intestine was isolated quickly,and sections of the intestine were cut at the duodenum,jejunum,ileocecal junction,colon and rectum for histopathological evaluation.ELISA was adopted to determine related inflammation factors while multi-color immunohistochemistry was used to calculate macrophage surface markers.The data was statistically analyzed.Results(1)Compared with the sham group,there was no significant difference in colon histology at 3 h and 6 h,but significant difference was detected in rectum scores only at 24 h.The scores of other intestinal segments were significantly different at each time point.The severity of ileocecal and colonic lesions after bleeding increased with time.The duodenum,jejunum and ileocecum were more critically injured at 3 h than the rectum at 6 h.The injury to the duodenum,jejunum,ileum and colon was much more pronounced than to the rectum at 12 h.(2)The expressions of TNF-α and IL-1β in the rectum were increased significantly at 12 h post operation.The expressions of IL-1β,TNF-α in the jejunum increased obviously at 3 h and 6 h,respectively.(3)Three hours after severe bleeding,the level of macrophages in the jejunum and ileocececal area increased significantly,and the percentage of M1 macrophages was higher.After 6 hours,the proportion of M2 macrophages in the jejunum and M1 macrophages decreased significantly.After 3 hours,the percentage of M1 macrophages in the colon decreased,but that of M2 macrophages increased.The proportion of M2 polarized macrophages in the duodenum and rectum increased at 3 h after severe bleeding but decreased at 6 h.Conclusion Pathological damage to intestinal sections after bleeding varies depending on the time,and is correlated with the inflammatory level of macrophages.

魏汉琪;王哲;于群;马骏;蒋兴伟;苏允琦;高锋华;宁畅文;安华英;巩家媛;刘鹏宇

军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室,北京 100850

基础医学

失血大鼠肠道炎症巨噬细胞

hemorrhagerat intestinal tractinflammationmacrophages

《军事医学》 2024 (11)

815-825,11

军事科学院军事医学研究院血液安全保障技术研究北京市重点实验室自主课题项目

10.7644/j.issn.1674-9960.2024.11.003

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