本文選題：熱休克預(yù)處理　切入點(diǎn)：HSF1　出處：《中南大學(xué)》2009年碩士論文

【摘要】： 細(xì)胞遷移發(fā)生于胚胎形成、機(jī)體免疫、急慢性炎癥及腫瘤等多種生理及病理過程中,是炎癥過程中必不可少的環(huán)節(jié)之一。炎癥發(fā)生時,白細(xì)胞可以通過血管壁滲出到血管外并在局部組織中聚集,這個過程稱為白細(xì)胞浸潤。單核/巨噬細(xì)胞通常是到達(dá)浸潤部位的主要白細(xì)胞之一。白細(xì)胞浸潤是一個極為復(fù)雜的連續(xù)過程,主要由白細(xì)胞的三個生物學(xué)特性決定:黏附作用、趨化作用、變形作用。其中趨化作用是白細(xì)胞參與炎癥反應(yīng)的重要前驅(qū)反應(yīng)。 熱休克反應(yīng)(HSR)是機(jī)體最重要的保護(hù)機(jī)制之一。熱休克因子1(HSF1)通過誘導(dǎo)熱休克蛋白(HSPs)的表達(dá),在細(xì)胞內(nèi)源性保護(hù)中發(fā)揮重要作用。近年國內(nèi)外研究發(fā)現(xiàn),HSF1及HSPs也參與了對小鼠胚胎成纖維細(xì)胞及人胚胎腎細(xì)胞遷移的調(diào)節(jié)過程。這為我們研究HSR及HSF1調(diào)控細(xì)胞遷移提供了重要信息。然而,HSR是否可以調(diào)控炎癥中重要細(xì)胞之一的巨噬細(xì)胞的遷移?HSF1是否及如何參與該調(diào)控過程?目前尚無相關(guān)研究。 為探討熱休克預(yù)處理對脂多糖(LPS)所致巨噬細(xì)胞遷移及黏附的影響,本研究首先采用腹腔注射大腸桿菌LPS的小鼠模型,通過加熱使小鼠肛溫升至42℃并維持15min制備熱休克預(yù)處理模型,采用支氣管肺泡灌洗觀察熱休克預(yù)處理對LPS所致小鼠肺泡白細(xì)胞滲出的影響;并采用LPS體外刺激的小鼠腹腔巨噬細(xì)胞及RAW264.7巨噬細(xì)胞株為模型,應(yīng)用趨化實(shí)驗(yàn)、劃痕愈合實(shí)驗(yàn)及黏附實(shí)驗(yàn)觀察熱休克預(yù)處理(42℃,1h)對LPS所致巨噬細(xì)胞遷移的影響;為探討HSF1在熱休克預(yù)處理對LPS所致巨噬細(xì)胞遷移的影響中是否發(fā)揮作用,采用HSF1基因敲除小鼠制備內(nèi)毒素血癥模型,應(yīng)用免疫組化技術(shù)觀察HSF1基因敲除小鼠肺、肝組織形態(tài)學(xué)變化及巨噬細(xì)胞浸潤情況;采用HSF1敲除小鼠腹腔巨噬細(xì)胞,應(yīng)用趨化實(shí)驗(yàn)觀察HSF1敲除對LPS所致巨噬細(xì)胞遷移的影響;并采用HSF1過表達(dá)的RAW264.7巨噬細(xì)胞株,應(yīng)用趨化實(shí)驗(yàn)觀察HSF1過表達(dá)對LPS所致巨噬細(xì)胞遷移的影響。 結(jié)果發(fā)現(xiàn):(1).LPS(12mg/kg,2h)腹腔注射促進(jìn)小鼠血清中TNF-α、IL-6、IL-10的釋放;LPS(600ng/ml,2h)促進(jìn)RAW264.7巨噬細(xì)胞株TNF-α的釋放。(2).腹腔注射LPS(12mg/kg,48h)導(dǎo)致小鼠肺泡灌洗液中白細(xì)胞總數(shù)及巨噬細(xì)胞數(shù)均明顯增加,熱休克預(yù)處理抑制了LPS所致小鼠白細(xì)胞及巨噬細(xì)胞向肺泡的遷移;采用體外趨化實(shí)驗(yàn)檢測LPS(600ng/ml,24h)刺激RAW264.7巨噬細(xì)胞株及小鼠腹腔巨噬細(xì)胞,發(fā)現(xiàn)兩種細(xì)胞的遷移均明顯增加,而熱休克預(yù)處理能抑制LPS所致的巨噬細(xì)胞遷移;LPS處理(600ng/ml,24h)促進(jìn)兩種巨噬細(xì)胞向劃痕部位的遷移而熱休克預(yù)處理可抑制LPS對巨噬細(xì)胞遷移的促進(jìn)作用;LPS處理(600ng/ml,1h)可使上述兩種巨噬細(xì)胞與細(xì)胞外基質(zhì)的黏附增加,而熱休克預(yù)處理可抑制LPS對巨噬細(xì)胞黏附的促進(jìn)作用。(3).HSF1基因敲除廢除了熱休克預(yù)處理對LPS所致的小鼠肺、肝組織巨噬細(xì)胞浸潤及組織損傷的保護(hù)作用,也廢除了熱休克預(yù)處理對LPS所致小鼠腹腔巨噬細(xì)胞遷移的抑制作用。(4).HSF1基因過表達(dá)無需進(jìn)行熱休克預(yù)處理即能抑制LPS所致的RAW264.7巨噬細(xì)胞遷移,而熱休克處理并不能進(jìn)一步增強(qiáng)這種抑制作用。 綜上所述,LPS在整體小鼠和離體巨噬細(xì)胞模型均能促進(jìn)巨噬細(xì)胞的遷移;熱休克預(yù)處理能抑制LPS所致的小鼠肺、肝組織損傷和巨噬細(xì)胞浸潤以及LPS所致離體巨噬細(xì)胞的遷移;HSF1參與了熱休克預(yù)處理對LPS所致巨噬細(xì)胞遷移的抑制作用。
[Abstract]:cell migration occurs in many physiological and pathological processes , such as embryonic development , organism immunity , acute and chronic inflammation and tumor , and is one of the necessary links in the process of inflammation .



Heat shock factor ( HSR ) is one of the most important protective mechanisms of HSR , which plays an important role in the endogenous protection of human embryonic kidney cells by inducing the expression of heat shock protein ( HSPs ) . In recent years , it has been found that HSFs and HSPs also participate in the regulation of the migration of mouse embryonic fibroblasts and human embryonic kidney cells .



In order to investigate the effects of heat shock pretreatment on the migration and adhesion of LPS - induced macrophages in mice , the effects of heat shock pretreatment on LPS - induced macrophage migration were studied by means of heat shock pretreatment .



The results showed that : ( 1 ) LPS ( 12 mg / kg , 2h ) could promote the release of TNF - 偽 , IL - 6 and IL - 10 in serum of mice . LPS ( 600ng / ml , 2h ) promoted the release of TNF - 偽 in RAW264.7 macrophage . LPS ( 600ng / ml , 24h ) stimulated the migration of macrophages and macrophages . LPS treatment ( 600ng / ml , 24h ) promoted the migration of two kinds of macrophages to scratch sites . LPS treatment ( 600ng / ml , 1h ) promoted the adhesion of two kinds of macrophages to the extracellular matrix . The effect of heat shock pretreatment on LPS - induced infiltration and tissue injury of mouse lung and liver tissue was abolished by HSFI , and the inhibitory effect of heat shock pretreatment on peritoneal macrophage migration in mice induced by LPS was also abolished . It is not necessary to perform heat shock pretreatment to inhibit the migration of RAW264.7 macrophages due to LPS , but the heat shock treatment can not further enhance the inhibition .



In conclusion , LPS can promote the migration of macrophages in both the whole mouse and the ex vivo macrophage model . Heat shock pretreatment can inhibit the migration of LPS - induced lung , liver tissue injury and macrophage infiltration and LPS - induced dissociation of macrophages .

【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 呂運(yùn)成;王佐;危當(dāng)恒;姜志勝;萬煒;李國華;童中藝;王貴學(xué);;基質(zhì)細(xì)胞衍生因子1α—CXCR4趨化THP-1細(xì)胞遷移及氧化型低密度脂蛋白的影響[J];中國動脈硬化雜志;2007年01期

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本文編號：1716322