首页|高原寒冷环境猪腹部肠管火器贯通伤肠道继发性通透性改变研究

高原寒冷环境猪腹部肠管火器贯通伤肠道继发性通透性改变研究

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目的 观察高原寒冷环境猪腹部肠管火器贯通伤后未直接损伤肠道继发性通透性改变及可能机制。方法 55只实验猪分为高原寒冷组(HC)及平原常温组(LN)共2个大组,每个大组下按照观察时间点将分为0 h、2 h、4 h、8 h、24 h 5个实验亚组,HC组下每个亚组6只实验猪,LN组下每个亚组5只实验猪。按照分组时间点安乐死实验猪,取肠道组织,以ELISA法检测肠道组织匀浆炎症因子TNF-α和IL-6的水平以及血液中肠道通透性标志蛋白二氨氧化酶(diarnine oxidse,DAO)、D-乳酸(D-lactate,D-LA)浓度;取 LN-0 h、LN-8 h 以及 HC-8 h 时间点实验猪肠道组织,使用HE染色后观察其肠道病理变化并评分,通过蛋白质印迹法(Western blot)检测肠道通透性相关的咬合蛋白(Occludin)、闭合小环蛋白1(zonula occluden-1,ZO-1)、闭合蛋白3(Claudin-3)与通道相关的Toll样受体 4(toll-like receptors 4,TLR4)、核因子 κB(nuclear factor-κB,NF-κB)、肌球蛋白轻链激酶(myocin light chain kinase,MLCK)的含量。结果 HC组与LN组伤后均导致了典型的腹部肠管贯通伤,腹腔感染评分及肠管粘连在两组间无明显差异(P>0。05)。HC组与LN组实验猪血清中DAO、D-LA随时间逐渐增加,HC组DAO、D-LA在全部时间点均显著高于LN组(P<0。01或P<0。001),HC组的DAO、D-LA增加最快的时间段为4~8 h而LN组则为8~24 h。HC组肠道组织病理评分显著高于LN组实验猪(P<0。01)。LN组与HC组肠道组织炎症因子TNF-α和IL-6随时间均不断升高,HC组炎症因子增加最明显的时间点为4~8 h,LN组为8~24 h。HC组实验猪肠道组织中的 IL-6、TNF-α 在全部时间中均高于 LN 组(P<0。05、P<0。01 或 P<0。001)。HC-8 h 组 Occludin 与 ZO-1 较 LN-8 h时间点均下降,差异具有统计学意义(P<0。05),而Claudin-3对比LN-8 h显著下降(P<0。001)。HC-8 h组TLR4、NF-κB与MLCK均高于LN-8h组,差异具有统计学意义(P<0。05)。结论 高原寒冷环境会导致腹部肠管火器贯通伤后继发性肠道通透性增加,其机制可能与TLR4/NF-κB/MLCK通路的激活相关。
Study on changes to intestinal permeability secondary to firearm-related penetrating wound of pig abdominal intestine in cold environment at high altitudes
Objective To observe the changes to,and possible mechanism of,intestinal permeability in pigs without direct injury after an abdominal-and intestinal-penetrating injury from firearms in cold environment at high altitudes.Methods Fifty-five experimental pigs were divided into two groups:high-altitude cold group(HC)and low-altitude normal temperature group(LN).According to the observation time,each group was divided into five experimental subgroups:0h,2h,4 h,8h,and 24 h.There were six pigs in each HC subgroup and five pigs in each LN subgroup.After euthanasia,intestinal tissues were taken,and the levels of the inflammatory factors TNF-α,and IL-6 in intestinal homogenate and the concentrations of intestinal permeability-related proteins DAO and D-lactate acid in blood were detected by ELISA method.The intestinal tissues of experimental pigs were taken at 0 h and 8 h for LN and 8 h for HC,and intestinal pathological changes were observed and scored after HE staining.The concentrations of Occludin,ZO-1,Claudin-3,TLR4,NF-κB,and MLCK(proteins related to intestinal permeability)were detected by Western blot to explore the effect of a cold environment at high altitude on secondary intestinal permeability changes after injury and the possible mechanisms.Results Both the HC group and LN group experienced typical abdominal intestinal penetrating injuries,and there were no significant differences in their abdominal infection scores or intestinal adhesion(P>0.05).The levels of DAO and D-LA in the serum of experimental pigs in the HC and LN groups gradually increased over time.The levels of DAO and D-LA in the HC group were significantly higher than those in the LN group at all time points(P<0.01 or P<0.001).The fastest increase in DAO and D-LA in the HC group was 4 h to 8 h,while in the LN group,it was 8 h to 24 h.The pathological score of intestinal tissue in the HC group was significantly higher than that in the LN group of experimental pigs(P<0.01).The inflammatory factors TNF-α and IL-6 both increased over time in the intestinal tissue of LN and HC groups.The most significant time point for a increase of inflammatory factors in the HC group was 4 h to 8 h,while in the LN group,it was 8 h to 24 h.The intestinal tissue IL-6 and TNF-α levels of experimental pigs in the HC group were higher than those in the LN group the entire time(P<0.05,P<0.01,or P<0.001).The levels of occludin and ZO-1 in the HC group at 8 h decreased significantly compared to those of the LN group at the 8 h time point(P<0.05),while claudin-3 showed a significant decrease in LN(P<0.001).In the HC group,TLR4,NF-κB,and MLCK were both higher than those in the LN group at 8 h,and the difference was statistically significant(P<0.05).Conclusions A high-altitude cold environment can lead to a secondary increase in intestinal permeability after abdominal-penetrating firearm injury,and its mechanism may be related to the TLR4/NF-κB/MLCK pathway.

cold environment at high altitudesabdominal firearm injuryintestinal permeabilitytight junction

屈金权、李佳佳、鲁洪男、杨欣悦、孙赳、梁飞行、王岩、刘江伟

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新疆军区总医院高原高寒医学保障重点实验室,乌鲁木齐 830000

新疆军区总医院新疆特殊环境医学重点实验室,乌鲁木齐 830000

新疆军区总医院临床检验中心,乌鲁木齐 830000

新疆公安厅特警总队训练基地,乌鲁木齐 830000

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高原寒冷环境 腹部火器伤 肠道通透性 紧密连接

实验动物专项课题

SYDW[2017]11号

2024

中国比较医学杂志
中国实验动物学会,中国医学科学院医学实验动物研究所

中国比较医学杂志

CSTPCD北大核心
影响因子:0.473
ISSN:1671-7856
年,卷(期):2024.34(9)