HSP70在百草枯诱导的肺部炎症反应中的表达

发布时间：2018-06-29 00:07

本文选题：百草枯中毒 + 热休克蛋白70　；参考：《中国医科大学》2010年硕士论文

【摘要】： HSP70在百草枯诱导的肺部炎症反应中的表达实验目的本研究将应用病理HE染色方法检测百草枯中毒大鼠肺组织病理变化及病理损伤程度,应用免疫组织化学及Western blot方法检测百草枯中毒大鼠肺组织中热休克蛋白(HSP70)的表达水平。探索百草枯致大鼠急性肺损伤的发生机制,对临床工作中百草枯中毒患者早期的救治提供有利的线索。实验方法 72只健康Wistar大鼠随机平均分成3组,分别腹腔注射生理盐水(A组)、20 mg/kg百草枯(B组)和40 mg/kg百草枯(C组)。各组动物1、2、3、6、12和24 h随机编入6个观察时点。造模前,各组大鼠禁食12 h,不禁水。A组大鼠腹腔注射1 ml生理盐水,B组和C组大鼠用生理盐水将PQ稀释至1 ml腹腔注射染毒。每组分别于6个观察时点用10％水合氯醛腹腔注射后处死4只大鼠,留取肺组织标本,左侧肺组织置于4％多聚甲醛中固定12 h,用于HE染色,免疫组织化学染色。取右侧肺组织放于-70℃冰箱备用,Western blot方法测定肺组织中HSP70的表达。实验结果 HE染色,染毒大鼠肺组织发生急性肺损伤、透明膜的形成和炎性细胞浸润等急性肺损伤典型的病理改变。免疫组织化学结果,B组中HSP70的表达于2 h开始升高,6 h达到峰值,24 h与A组无差别。C组中HSP70的表达于1 h开始升高,3 h达到峰值,24 h与A组无差别。且C组2、3和12 h的表达均显著高于B组。Western blot结果,B组大鼠2、3、6 h时点HSP70的表达在肺组织中较A组明显增强,其他时间点与A组比无明显差异。C组1、2、3、6、12 h时点HSP70的表达较A组均明显增强,2、3、12 h时点HSP70的表达较B组明显增强。结论百草枯诱导的急性肺损伤可引起支气管、细支气管和肺泡上皮细胞HSP70应激性表达,中毒剂量增大,肺损伤加重,HSP70的表达亦随之增强,而肺组织的病理损伤却没有显著性差异,提示HSP70对肺损伤起保护作用。
[Abstract]:Expression of HSP70 in paraquat induced pulmonary inflammation objective to study the pathological changes and degree of lung injury in paraquat poisoned rats by pathological HE staining. The expression of heat shock protein (HSP70) in the lung tissue of paraquat poisoned rats was detected by immunohistochemistry and Western blot. To explore the mechanism of acute lung injury induced by paraquat in rats and to provide a useful clue for the early treatment of paraquat poisoning in clinical work. Methods Seventy-two healthy Wistar rats were randomly divided into 3 groups and received intraperitoneal injection of normal saline (group A) for 20 mg/kg paraquat (group B) and 40 mg/kg paraquat (group C). The animals in each group were randomly assigned to 6 observation time points at 12 and 24 hours. Before the model was established, rats in each group fasted for 12 hours. Rats in group A were injected intraperitoneally with 1 ml normal saline and rats in group C diluted PQ to 1 ml by intraperitoneal injection. Four rats were killed after intraperitoneal injection of 10% chloral hydrate at 6 observation points in each group. The left lung tissue was fixed in 4% paraformaldehyde for 12 hours and was used for HE staining and immunohistochemical staining. The expression of HSP70 in lung tissue was determined by Western blot method. The results showed that acute lung injury, hyaline membrane formation and inflammatory cell infiltration were typical pathological changes in lung tissue of rats exposed to HE. Immunohistochemical results showed that the expression of HSP70 in group B began to increase at 2 h and reached a peak at 6 h. There was no difference in HSP70 expression between group A and group A. the expression of HSP70 in group C increased at 1 h and reached peak value at 3 h. There was no difference between group A and group A at 24 h. The expression of HSP70 in group C was significantly higher than that in group B at 2h and 12h. The results of Western blot showed that the expression of HSP70 in group B was significantly higher than that in group A. There was no significant difference between other time points and A group. The expression of HSP70 in group C was significantly higher than that in group A at 12 h after 12 h. The expression of HSP70 in group C was significantly higher than that in group B at 12 h. Conclusion the acute lung injury induced by paraquat can induce the stress expression of HSP70 in bronchioles, bronchioles and alveolar epithelial cells. However, there was no significant difference in pathological injury of lung tissue, suggesting that HSP70 plays a protective role in lung injury.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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