本文選題：胰腺炎 + 自噬�。� 參考：《青島大學》2017年碩士論文

【摘要】：目的:在重癥急性胰腺炎(severe acute pancreatitis,SAP)中,腸源性感染是SAP患者后期感染的重要預后指標,細菌移位(bacterial translocation,BT)是腸源性感染的重要原因,在SAP早期便可引起全身炎癥反應綜合征(systemic inflammatory response syndrome,SIRS),進而導致多器官功能衰竭(multiple organ dysfunction syndrome,MODS)與膿毒血癥,是SAP后期死亡的主要因素,而SAP腸粘膜屏障損傷發(fā)生BT的具體分子機制尚不明確。自噬參與多種疾病的病理生理過程,腸上皮自噬對于腸道穩(wěn)態(tài)及腸道免疫發(fā)揮重要作用,有研究證實腸上皮自噬具有清除侵入機體腸道細菌的作用,并且近期體外研究表明,自噬增強腸上皮緊密連接(tight junction,TJ)蛋白的表達。但是腸上皮自噬對于SAP患者腸道BT影響尚不明確,我們旨在研究SAP中腸上皮自噬對于腸道BT與腸上皮TJ表達的影響。方法:根據(jù)2012年修訂的亞特蘭大急性胰腺炎分類定義標準,并給予急性胰腺炎患者急性生理與慢性健康評分(APACHE-Ⅱ)評分,選取發(fā)病24小時內(nèi)入院APACHE-Ⅱ評分在8到12分之間的31位SAP患者。根據(jù)SAP患者外周血中,基于16SrDNA測序結(jié)果,確定患者外周血中細菌DNA的有無,將SAP患者分為細菌移位陽性組與細菌移位陰性組即BT(+)和BT(-),另選取健康的8人作為健康對照組(healthy control,HC),同時使用結(jié)腸鏡獲取SAP患者與健康對照組的結(jié)腸上皮粘膜組織;采用酶聯(lián)免疫吸附法(ELISA)測定血清內(nèi)毒素水平;采用免疫印跡技術(shù)(Western blot)檢測腸上皮組織的緊密連接蛋白Zonula occluden-1(ZO-1)、claudins-2(CL-2)、occludin(OC)以及自噬標志蛋白微管相關(guān)蛋白1輕鏈3Ⅱ(LC3Ⅱ)的表達水平。結(jié)果:在納入的31位SAP患者中,有14位患者外周血樣本可檢測到細菌DNA,陽性率為45.2%,BT(+)有14人和BT(-)有17人。經(jīng)方差分析結(jié)果顯示,血清內(nèi)毒素在三組間差異具有統(tǒng)計學意義(F=6.981,P0.05),利用LSD-t檢驗比較結(jié)果顯示,在SAP患者中BT(+)組中檢測到血清內(nèi)毒素水平顯著高于BT(-)組和HC組(t=4.973,P0.05,t=8.661,P0.05);CL-2蛋白表達在三組間的差異具有統(tǒng)計學意義(F=8.641,P0.05),利用LSD-t進行組間比較結(jié)果顯示,在BT(+)組患者中CL-2蛋白表達水平高于BT(-)組和HC組(t=6.875,P0.05,t=10.733,P0.05);BT(+)組患者的TJ蛋白ZO-1和OC均低于BT(-)組患者。此外,自噬相關(guān)蛋白LC3Ⅱ表達在三組間的差異具有統(tǒng)計學意義(F=11.574,P0.05),BT(-)患者腸上皮LC3Ⅱ表達高于BT(+)患者(t=4.765,P0.05,t=6.981,P0.05),且在SAP患者中CL-2與LC3Ⅱ表達呈負相關(guān)(r=-0.71,P0.05),相反的是,ZO-1和OC與LC3Ⅱ表達呈正相關(guān)(r=0.79,P0.05;r=0.88,P0.05)。結(jié)論:在SAP患者中腸上皮自噬是激活的,腸上皮自噬激活可能減少細菌移位的發(fā)生,其機制可能是腸上皮自噬影響腸上皮緊密連接作用,增強腸上皮TJ作用減少細菌移位的發(fā)生。
[Abstract]:Objective: in severe acute patients with severe acute pancreatitis (SAP), enterogenous infection is an important prognostic indicator of late infection in SAP patients, and bacterial translocation is an important cause of enterogenic infection. Systemic inflammatory response syndrome and multiple organ dysfunction syndrome (mods) and sepsis can be caused in early stage of SAP, which is the main cause of death in the later stage of SAP. However, the molecular mechanism of BT in SAP intestinal barrier injury is still unclear. Autophagy is involved in the pathophysiological process of many diseases. Intestinal epithelium autophagy plays an important role in intestinal homeostasis and intestinal immunity. Autophagy enhances the expression of tight junction TJ protein in intestinal epithelium. However, the effect of intestinal epithelial autophagy on intestinal BT in patients with SAP is not clear. We aim to study the effect of SAP midgut epithelium autophagy on the expression of BT and TJ in intestinal epithelium. Methods: according to the Atlanta acute pancreatitis classification standard revised in 2012, and the acute physiological and chronic health score (APACHE- 鈪，

本文編號：1813880