本文關(guān)鍵詞： 白細(xì)胞介素36α 膿毒血癥 宿主應(yīng)答 巨噬細(xì)胞　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：背景膿毒血癥是由感染引起的全身炎癥反應(yīng)綜合征,這種由感染引起的宿主應(yīng)答調(diào)節(jié)紊亂最終會導(dǎo)致器官功能的紊亂,嚴(yán)重威脅生命。膿毒血癥的死亡率很高,且目前臨床對膿毒血癥缺乏有效的治療措施。近年來的研究數(shù)據(jù)提示,白細(xì)胞介素-1(Interleukin-1,IL-1)家族的新成員白細(xì)胞介素36α(Interleukin-36α,IL-36α)在炎癥疾病的病理機(jī)制中占有重要地位,但其在膿毒血癥中對宿主應(yīng)答的作用尚未得知。方法用盲腸結(jié)扎穿刺(CLP)的方法構(gòu)建膿毒血癥模型,誘導(dǎo)多細(xì)菌感染的膿毒血癥,以研究IL-36α在膿毒血癥中對宿主應(yīng)答的影響;用定量PCR和ELISA檢測IL-36α在小鼠體內(nèi)的表達(dá)量;通過進(jìn)行生存率試驗和細(xì)菌載量的測定,來衡量IL-36α在膿毒血癥中的保護(hù)或損傷作用;取小鼠肺、肝、脾、腎組織做組織切片并做HE染色,分析組織內(nèi)的炎性改變,通過TUNEL實驗評判細(xì)胞凋亡程度;用流式細(xì)胞術(shù)和瑞士-吉姆薩染色來分析腹腔灌洗液中有核細(xì)胞的數(shù)量及種類;提取腹腔巨噬細(xì)胞及骨髓中性粒細(xì)胞進(jìn)行體外實驗,檢測IL-36α對小鼠巨噬細(xì)胞和中性粒細(xì)胞的功能的影響,取細(xì)胞上清測定IL-36α對炎癥細(xì)胞因子表達(dá)的影響。結(jié)果IL-36α在膿毒血癥中的表達(dá)顯著上調(diào)。IL-36α處理過的CLP重癥膿毒血癥模型小鼠的死亡率降低。與PBS對照組小鼠相比,IL-36α處理組小鼠表現(xiàn)出更有效的細(xì)菌清除能力,組織炎癥程度被抑制,器官損傷更輕,且免疫細(xì)胞凋亡減少。IL-36α在膿毒血癥中還可能具有治療價值,這在使用IL-36α抗體的實驗中也得到了證實,特異性地封閉體內(nèi)IL-36α導(dǎo)致在輕度膿毒血癥中小鼠的死亡率增加。除此之外,我們還發(fā)現(xiàn)IL-36α增強(qiáng)了巨噬細(xì)胞對細(xì)菌的吞噬和殺傷,從而使得局部和全身的細(xì)菌清除能力增強(qiáng)。更重要的是,在構(gòu)建膿毒血癥模型之前耗盡小鼠體內(nèi)巨噬細(xì)胞,IL-36α的這種保護(hù)作用就消失了。結(jié)論我們的結(jié)果表明IL-36α在宿主對膿毒血癥的防御應(yīng)答中發(fā)揮了重要的保護(hù)作用,并提示IL-36α對膿毒血癥具有潛在的治療效果。
[Abstract]:Background sepsis is a systemic inflammatory response syndrome caused by infection. The host response regulation disorder caused by infection can eventually lead to organ dysfunction, which is a serious threat to life. The mortality rate of sepsis is very high. Recent studies indicate that interleukin-36 偽 (IL-36 偽), a new member of the Interleukin-36 偽 (IL-36 偽) family, plays an important role in the pathological mechanism of inflammatory diseases. Methods the sepsis model was established by cecal ligation and puncture to induce sepsis caused by bacterial infection, in order to study the effect of IL-36 偽 on host response in sepsis. Quantitative PCR and ELISA were used to detect the expression of IL-36 偽 in mice. Survival test and bacterial load were used to evaluate the protective or injury effect of IL-36 偽 in sepsis. Renal tissue sections and HE staining were used to analyze the inflammatory changes in the tissues and the degree of apoptosis was evaluated by TUNEL assay, and the number and types of nucleated cells in peritoneal lavage fluid were analyzed by flow cytometry and Swiss Gimsa staining. Peritoneal macrophages and bone marrow neutrophils were extracted to investigate the effects of IL-36 偽 on the function of mouse macrophages and neutrophils. The effect of IL-36 偽 on the expression of inflammatory cytokines was measured by supernatant. Results the expression of IL-36 偽 in sepsis was significantly up-regulated. The mortality of CLP model mice treated with IL-36 偽 was decreased. Compared with the control group of PBS mice, the mortality of IL-36 偽 was significantly decreased. The mice in the treatment group showed more effective bacterial clearance. Tissue inflammation is inhibited, organ damage is lighter, and reduced apoptosis of immune cells. IL-36 偽 may also have therapeutic value in sepsis, which has also been confirmed in the use of IL-36 偽 antibody. Specific blocking of IL-36 偽 in vivo leads to an increase in mortality in mice with mild sepsis. In addition, we found that IL-36 偽 enhanced phagocytosis and killing of bacteria by macrophages. Which increases the ability to remove bacteria locally and throughout the body. More importantly, The protective effect of IL-36 偽 on macrophage in mice was disappeared before the model of sepsis was established. Conclusion our results suggest that IL-36 偽 plays an important protective role in the host's defense against sepsis. The results suggest that IL-36 偽 has potential therapeutic effect on sepsis.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R459.7

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