劳力型热射病大鼠炎症反应中细胞因子的研究

发布时间：2018-01-27 09:54

本文关键词： 热射病动物模型炎症反应细胞因子　出处：《中国人民解放军医学院》2014年硕士论文　论文类型：学位论文

【摘要】：目的：本研究旨在通过建立经典型和劳力型热射病大鼠模型，比较其在同样的环境条件下发病严重程度及炎症反应中细胞因子水平的变化。方法：建立经典型与劳力型热射病大鼠模型，雄性SD大鼠随机分为空白对照组、高温高湿安静暴露组（Classical Heat Stroke,CHS组）、高温高湿跑台运动组（Exertional Heat Stroke, EHS组），每组10只。（1）监测大鼠肛温（T，℃），记录达到发病终点T42.0℃的时间；（2）发病后取大鼠血清送至我院生化科，检测肌酸激酶（creatine kinase, CK）、天冬氨酸氨基转移酶（aspartate aminotransferase,AST）、谷氨酸转氨酶（glutamate transaminase, ALT）、总胆红素（total bilirubin, TB）及直接胆红素（direct bilirubin, DB）水平；（3）观察各组大鼠解剖后主要脏器病理变化（Hematoxylin-eosin染色）。观察各组大鼠至发病终点，用10%的水合氯醛1ml腹腔内注射将实验动物麻醉后留取大鼠各主要脏器病理标本（小肠、肾脏、肝脏、肺脏、心脏、骨骼肌），采用HE染色后观察各标本病理改变。采用免疫酶联反应方法（ELISA）检测各组大鼠血清白介素-1β（Interleukin-1β，IL-1β）、白介素-6（Interleukin-6，IL-6）、白介素-8（Interleukin-8，IL-8）、白介素-10（Interleukin-10，IL-10）、肿瘤坏死因子α（Tumor necrosis factor-α，TNF-α）、内毒素（LPS）、高迁移率组蛋白1（High mobility group box-1protein,HMGB-1）浓度，比较经典型与劳力型热射病大鼠模型中各细胞因子达到峰浓度时间及峰值的不同。结果：首先，我们成功建立了经典型与劳力型热射病大鼠模型。各组大鼠发病平均时间为：EHS组（58.333±9.309）min，CHS组为（108.333±23.166）min，EHS组较CHS组发病时间明显加快（P0.01）。EHS组与CHS组大鼠发病后血清CK水平的平均值分别为（4444.44±1287.915）U/L、（1208.217±192.802）U/L，EHS组较CHS组血清CK水平明显增高（P0.01）；实验两组大鼠发病时AST水平的平均值分别为：CHS组（270.043±36.999）U/L、EHS组（198.486±21.853）U/L，发病2h时CHS与EHS两组分别为（460.900±143.300）U/L、（386.133±32.331）U/L，发病4h时CHS与EHS两组分别为（704.000±75.003）U/L、（538.467±60.761）U/L，发病6h时CHS与EHS两组分别为（868.433±75.394）U/L、（791.433±78.119）U/L；实验两组大鼠发病时ALT水平的平均值分别为：CHS组（90.400±35.055）U/L、EHS组（61.850±11.865）U/L，发病2h时CHS与EHS两组分别为（149.167±7.156）U/L、（74.5±23.998）U/L，，发病4h时CHS与EHS两组分别为（343.200±32.045）U/L、（131.767±27.543）U/L，发病6h时CHS与EHS两组分别为（337.200±17.801）U/L、（555.567±68.148）U/L；各组大鼠血清TB、DB水平测量无统计学意义；各组大鼠凝血指标在发病时变化最明显，随着发病时间延长差异不明显。本研究还发现EHS组大鼠各主要器官（小肠、肾脏、骨骼肌等）病理损伤较CHS组大鼠明显加重。其次，IL-1β在发病1h、2h、3h、4h、6h及8h与对照组及CHS组均有显著差异；而IL-6仅在发病当时高于CHS及对照组，随着时间发展下降明显，甚至低于对照组；在6个观察时间点中，热暴露结束后2h内毒素水平最高，并且与正常对照比较,热暴露结束后2h内毒素水平显著升高，具有统计学差异，p=0.013；CHS组与EHS组TNF-α水平在发病当时均高于对照组，且在发病当时EHS组TNF-α水平高于CHS组；CHS组与EHS组HMGB1水平在发病当时均高于对照组，且在发病当时EHS组HMGB1水平高于CHS组；其余细胞因子IL-8、IL-10与对照组相比呈现升高趋势，与经典型热射病大鼠模型相比差异无统计学意义。结论：（1）在动物模型上，劳力型热射病较经典型热射病在发病初期的各器官功能损伤发展更迅速，更严重；（2）IL-6同时发挥炎症及抗炎的双刃剑作用，推测在EHS发病过程中IL-6可能发挥着抗炎作用；IL-1β可能是EHS致炎的主要细胞因子；HMGB1在热射病发病早期即可升高，并与其他致炎因子相互促进分泌。
[Abstract]:Objective : The aim of this study was to compare the changes of cytokine levels in severity and inflammatory response in the same environmental conditions by establishing a model of classic and labor - type heat - injection rats . Methods : A typical model of male SD rats was established . Male SD rats were randomly divided into two groups : blank control group , high - temperature high - humidity and quiet exposure group ( CHS group ) , high - temperature and high - humidity treadmill exercise group ( EHS group ) , 10 rats in each group . Serum levels of interleukin - 1尾 ( IL - 1尾 ) , aspartate aminotransferase ( AST ) , aspartate aminotransferase ( AST ) , aspartate aminotransferase ( AST ) , glutamate transaminase ( ALT ) , total bilirubin ( total bilirubin , TB ) and direct bilirubin ( DB ) were observed . High mobility group box - 1protein ( HMGB - 1 ) was used to compare the peak concentration time and peak value of each cytokine in a typical model of stress - type heat injection . Results : The mean time of serum CK in the group of CHS and EHS in the group of CHS and EHS was ( 416.167 卤 7.156 ) U / L , ( 158.486 卤 21.853 ) U / L , ( 138.486 卤 23.998 ) U / L , ( 138.486 卤 23.998 ) U / L , and the CHS and EHS in group CHS and EHS respectively were ( 347.200 卤 7.156 ) U / L and ( 71.433 卤 78.119 ) U / L , respectively . At the time of 6 observation time , the endotoxin level was highest at 2h after the end of heat exposure , and the level of TNF - 伪 in the group of CHS and EHS was higher than that of the control group at the time of the onset of heat exposure , and the levels of TNF - 伪 in the group of CHS and EHS were higher than those in the control group . Conclusion : ( 1 ) In the animal model , it is more rapid and more serious to develop the function of IL - 6 in the early stage of the disease . ( 2 ) IL - 6 may play an anti - inflammatory effect in the pathogenesis of EHS . IL - 1尾 may play an anti - inflammatory effect in the pathogenesis of EHS ; the IL - 1尾 may be the main cytokine of EHS - induced inflammation .

【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R594.11

【共引文献】