TLR-9相关因子在急性肺挫伤中的表达及意义

发布时间：2018-03-03 06:08

本文选题：急性肺挫伤　切入点：TLR-9受体　出处：《贵州医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:Toll受体9(Toll-like receptors-9,TLR-9)在胸部创伤所致大鼠急性肺挫伤中表达的意义。方法:选取2016年4月~5月购自贵州医科大学动物实验中心的40只健康、雄性SD(Sprague-Dawley,SD)大鼠作为研究对象。利用小型生物撞击机(由第三军医大学予以提供),在500kpa下对大鼠胸部进行撞击,形成重度肺挫伤。参照AIS-90标准,对受伤后大鼠进行评分,大鼠受伤后会出现频繁、深沉的呼吸,较之前更明显,呼吸急促、口唇发钳、苏醒延迟、精神萎靡、食欲减退、活动减少。上述提示:代谢性呼吸增快在大鼠身上已经出现,目的在于适应氧供。因此,采用500kpa撞击较好的模拟了大鼠急性肺挫伤。随机分为5组,每组8只:正常对照组;胸部创伤致急性肺挫伤模拟组(500kpa)撞击后5,24,48,72小时,于股动脉放血处死。取大鼠血清及肺组织。HE染色观察大鼠肺组织病理学改变、肺组织湿/干质量比、采用免疫组织化学法、Elisa法检测血清及肺泡灌洗液中TLR-9受体的蛋白表达水平、予RT-PCR(Reverse transcription-PCR,RT-PCR)法检测肺组织TLR-9、IL-6(Interleukin-6,IL-6)的mRNA表达水平;Western blot法检测肺组织TLR-9的蛋白水平。结果:肺组织病理学检查显示:与空白对照组比较,胸部创伤组大鼠肺水肿、肺组织充血、中性粒细胞浸润、炎性渗出较明显。免疫组织化学法回示:胸部创伤组大鼠光镜下显色呈明显棕黄色,空白对照组大鼠光镜下未见黄色,其中以胸部创伤48小时组大鼠染色最为明显。胸部创伤组大鼠肺组织湿/干质量比:胸部创伤组大鼠与空白对照组大鼠相比,胸部创伤组大鼠肺组织湿/干质量比明显低于空白对照组。ELISA:血清及肺泡灌洗液中TLR-9、IL-6表达水平,胸部创伤组高于对照组,差异有统计学意义(P0.05)。RT-PCR回示:胸部创伤组肺组织中TLR-9 mRNA、IL-6 mRNA的表达较空白对照组升高,差异有统计学意义(P0.05)。Western blot回示:胸部创伤组肺组织中TLR-9蛋白水平较空白对照组升高,差异有统计学意义(P0.05)。结论:大鼠创伤后TLR-9表达的增高,提示TLR-9参加了肺挫伤炎症反应的过程。
[Abstract]:Objective: to investigate the expression of Toll-like receptor 9 (TLR-9) in acute lung contusion induced by thoracic trauma in rats. Methods: from April 2016 to May, 40 healthy rats were selected from the Animal Experimental Center of Guizhou Medical University. Sprague-Dawley SD rats were used as the study subjects. Using a small biological impact machine (provided by the third military Medical University), the chest of the rats was impacted at 500kpa, resulting in severe lung contusion. According to AIS-90 criteria, the injured rats were graded. There are frequent, deep breaths in rats after injury, more pronounced than before, shortness of breath, lip forceps, delayed recovery, depression of mind, loss of appetite, and decreased activity. The aim was to adapt to oxygen supply. Therefore, acute lung contusion was simulated by 500kpa impact in rats. The rats were randomly divided into 5 groups: normal control group (n = 8), acute lung contusion simulation group induced by chest trauma (n = 500 kpa) and control group (n = 8). The pathological changes of lung tissue were observed by HE staining in serum and lung tissue. The wet / dry weight ratio of lung tissue was measured. The expression of TLR-9 receptor protein in serum and alveolar lavage fluid was detected by immunohistochemistry. The expression level of mRNA in lung tissue was detected by RT-PCR(Reverse transcription-PCRD-PCRRT) and the protein level of TLR-9 in lung tissue was detected by Western blot. Results: compared with the control group, pulmonary edema and congestive lung tissue were detected by RT-PCR(Reverse transcription-PCRRT. Neutrophil infiltration and inflammatory exudation were obvious. The results of immunohistochemistry showed that the rats of thoracic trauma group showed obvious brownish yellow under light microscope, but no yellow was found in blank control group under light microscope. The wet / dry weight ratio of lung tissue in the thoracic trauma group was higher than that in the blank control group, and the wet / dry weight ratio of lung tissue in the thoracic trauma group was higher than that in the blank control group. The wet / dry weight ratio of lung tissue in the thoracic trauma group was significantly lower than that in the blank control group. The expression of TLR-9P IL-6 in serum and alveolar lavage fluid in the thoracic trauma group was significantly lower than that in the control group, and that in the chest trauma group was higher than that in the control group. The expression of TLR-9 mRNA-IL-6 mRNA in lung tissue of thoracic trauma group was higher than that of blank control group, and the level of TLR-9 protein in lung tissue of thoracic trauma group was higher than that of blank control group. Conclusion: the increase of TLR-9 expression in rats after trauma suggests that TLR-9 participates in the process of pulmonary contusion inflammation.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R655.3

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