本文選題：LPS + 細(xì)胞因子��； 參考：《中南大學(xué)》2006年博士論文

【摘要】：本研究從調(diào)控炎癥介質(zhì)基因表達(dá)的細(xì)胞內(nèi)信號轉(zhuǎn)導(dǎo)通路，探討熱休克反應(yīng)(heat shock response，HSR)以及HSP70抑制LPS所致細(xì)胞因子表達(dá)的機(jī)制。 經(jīng)采用RT-PCR、ELISA檢測發(fā)現(xiàn)，大腸桿菌脂多糖(LPS)可誘導(dǎo)RAW264.7小鼠巨噬細(xì)胞產(chǎn)生大量TNF-α，IL-1β和IL-15等細(xì)胞因子。當(dāng)細(xì)胞經(jīng)受熱休克反應(yīng)(42.5℃1h，恢復(fù)12h)后再進(jìn)行LPS刺激，，則LPS誘導(dǎo)的細(xì)胞因子表達(dá)受到抑制。在此基礎(chǔ)上采用過表達(dá)HSP70的RAW264.7細(xì)胞進(jìn)行實驗，發(fā)現(xiàn)HSP70同樣抑制了LPS誘導(dǎo)的細(xì)胞因子表達(dá)。 鑒于細(xì)胞內(nèi)信號轉(zhuǎn)導(dǎo)通路在調(diào)控炎癥介質(zhì)基因表達(dá)中的作用，本實驗采用磷酸化抗體檢測MAPK信號通路的活化，分析了HSR以及HSP70對LPS所致MAPK信號通路活化的影響。Western blot結(jié)果顯示，HSR以及HSP70自身均不影響MAPK的活化(磷酸化)。在LPS刺激30min后，HSR組以及HSP70過表達(dá)組中的JNK、ERK、p38的活化(磷酸化)程度與非HSR正常細(xì)胞組或轉(zhuǎn)空載體細(xì)胞組比較均無明顯區(qū)別。EMSA結(jié)果顯示，HSR以及HSP70不影響LPS刺激所致的MAPK下游轉(zhuǎn)錄因子AP-1的DNA結(jié)合活性。據(jù)此我們認(rèn)為，HSR以及HSP70不影響LPS所致的MAPK信號通路活化，其抑制LPS所致細(xì)胞因子表達(dá)的機(jī)制可能涉及其他因素。 由于NF-κB／IκB信號轉(zhuǎn)導(dǎo)通路的活化在LPS所致炎癥反應(yīng)的發(fā)生中具有重要作用，我們進(jìn)一步分析了HSP70過表達(dá)對LPS所致NF-κB信號通路活化的影響。通過免疫熒光、免疫細(xì)胞化學(xué)及Western blot
[Abstract]:The aim of this study was to investigate the mechanism of heat shock response to heat shock response (HSRs) and the inhibitory effect of HSP70 on the expression of cytokines induced by LPS by regulating the intracellular signal transduction pathway of inflammatory mediators gene expression. Elisa assay showed that Escherichia coli lipopolysaccharide (LPS) could induce the production of a large number of cytokines such as TNF- 偽, IL-1 尾 and IL-15 in macrophages of RAW264.7 mice. The expression of cytokines induced by LPS was inhibited when the cells were stimulated by LPS after heat shock reaction at 42.5 鈩

本文編號：1841177