本文選題：百草枯中毒 + 熱休克蛋白70��； 參考：《中國(guó)醫(yī)科大學(xué)》2010年碩士論文

【摘要】： HSP70在百草枯誘導(dǎo)的肺部炎癥反應(yīng)中的表達(dá) 實(shí)驗(yàn)?zāi)康?本研究將應(yīng)用病理HE染色方法檢測(cè)百草枯中毒大鼠肺組織病理變化及病理?yè)p傷程度,應(yīng)用免疫組織化學(xué)及Western blot方法檢測(cè)百草枯中毒大鼠肺組織中熱休克蛋白(HSP70)的表達(dá)水平。探索百草枯致大鼠急性肺損傷的發(fā)生機(jī)制,對(duì)臨床工作中百草枯中毒患者早期的救治提供有利的線索。 實(shí)驗(yàn)方法 72只健康Wistar大鼠隨機(jī)平均分成3組,分別腹腔注射生理鹽水(A組)、20 mg/kg百草枯(B組)和40 mg/kg百草枯(C組)。各組動(dòng)物1、2、3、6、12和24 h隨機(jī)編入6個(gè)觀察時(shí)點(diǎn)。造模前,各組大鼠禁食12 h,不禁水。A組大鼠腹腔注射1 ml生理鹽水,B組和C組大鼠用生理鹽水將PQ稀釋至1 ml腹腔注射染毒。每組分別于6個(gè)觀察時(shí)點(diǎn)用10％水合氯醛腹腔注射后處死4只大鼠,留取肺組織標(biāo)本,左側(cè)肺組織置于4％多聚甲醛中固定12 h,用于HE染色,免疫組織化學(xué)染色。取右側(cè)肺組織放于-70℃冰箱備用,Western blot方法測(cè)定肺組織中HSP70的表達(dá)。 實(shí)驗(yàn)結(jié)果 HE染色,染毒大鼠肺組織發(fā)生急性肺損傷、透明膜的形成和炎性細(xì)胞浸潤(rùn)等急性肺損傷典型的病理改變。免疫組織化學(xué)結(jié)果,B組中HSP70的表達(dá)于2 h開(kāi)始升高,6 h達(dá)到峰值,24 h與A組無(wú)差別。C組中HSP70的表達(dá)于1 h開(kāi)始升高,3 h達(dá)到峰值,24 h與A組無(wú)差別。且C組2、3和12 h的表達(dá)均顯著高于B組。Western blot結(jié)果,B組大鼠2、3、6 h時(shí)點(diǎn)HSP70的表達(dá)在肺組織中較A組明顯增強(qiáng),其他時(shí)間點(diǎn)與A組比無(wú)明顯差異。C組1、2、3、6、12 h時(shí)點(diǎn)HSP70的表達(dá)較A組均明顯增強(qiáng),2、3、12 h時(shí)點(diǎn)HSP70的表達(dá)較B組明顯增強(qiáng)。 結(jié)論 百草枯誘導(dǎo)的急性肺損傷可引起支氣管、細(xì)支氣管和肺泡上皮細(xì)胞HSP70應(yīng)激性表達(dá),中毒劑量增大,肺損傷加重,HSP70的表達(dá)亦隨之增強(qiáng),而肺組織的病理?yè)p傷卻沒(méi)有顯著性差異,提示HSP70對(duì)肺損傷起保護(hù)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.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R363

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本文編號(hào)：2079949