【摘要】：目的:p38絲裂原活化蛋白激酶(p38MAPK)信號通路是哺乳動物細胞中調(diào)控炎癥、細胞增殖與凋亡等最重要的信號通路之一。p38MAPK在細胞內(nèi)可被周圍環(huán)境中的應激刺激、炎性因子等激活,在缺血再灌注損傷的發(fā)生和發(fā)展過程中起到非常重要的作用。本文旨在通過探討雌二醇(Estradiol,E2)對皮瓣內(nèi)p38MAPK信號通路的影響及p38MAPK信號通路在皮瓣缺血再灌注損傷中的作用,為雌二醇治療皮瓣缺血再灌注損傷提供理論依據(jù)。方法:Wistar大鼠48只,雄性,12~14周,在無特定病原菌級(SPF級)實驗室條件下,建立大鼠腹部皮瓣缺血再灌注損傷模型。隨機將大鼠分為對照組(Ⅰ組):皮瓣切取后不作缺血再灌注處理,即刻原位縫合皮瓣;缺血再灌注組(Ⅱ組):切取皮瓣后,用無損傷顯微血管夾夾閉腹壁淺動靜脈6h后取出血管夾,確認腹壁淺動靜脈血流恢復后,同法縫合皮瓣;生理鹽水組(Ⅲ組):同Ⅱ組建立皮瓣模型后,腹腔注射生理鹽水;E2組(Ⅳ組):同Ⅱ組建立皮瓣模型,腹腔注射E2。術后觀察各組皮瓣一般情況;對皮瓣進行攝像,采用Image-Pro Plus.V6.0圖像分析系統(tǒng)計算皮瓣存活率;抽取皮瓣蒂部腹壁淺靜脈血,檢測血清中性粒細胞(neutrophils,NEU)數(shù)量、TNF-α及IL-10濃度;切取皮瓣行組織學觀察,應用免疫組織化學染色及Western Blot法檢測p38 MAPK及絲裂原活化蛋白激酶磷酸酶-2(MKP-2)的表達情況。結(jié)果:術后7d,大鼠皮瓣存活率Ⅳ組顯著高于Ⅱ、Ⅲ組(P0.05)。Ⅳ組蒂部靜脈血NEU數(shù)量及TNF-α濃度明顯低于Ⅱ、Ⅲ組(P0.05),而IL-10濃度顯著高于Ⅱ、Ⅲ組(P0.05)。皮瓣組織學與超微結(jié)構檢查結(jié)果顯示:與Ⅱ組、Ⅲ組相比,Ⅳ組皮瓣炎性滲出明顯較少,組織結(jié)構完整,細胞變性、壞死現(xiàn)象少見,細胞內(nèi)細胞器更加完整。Western Blot結(jié)果與免疫組化結(jié)果一致,顯示:Ⅱ、Ⅲ組p38MAPK表達明顯高于Ⅰ組(P0.05),而Ⅳ組p38MAPK表達較Ⅱ、Ⅲ組顯著降低(P0.05)。Ⅳ組MKP-2表達較Ⅰ、Ⅱ、Ⅲ組均顯著增加(P0.05)。相關性結(jié)果顯示:皮瓣存活率與NEU數(shù)量、TNF-α濃度之間均顯著負相關。結(jié)論:1.雌二醇可明顯抑制皮瓣內(nèi)NEU介導的炎癥級聯(lián)反應,減少TNF-α等促炎介質(zhì)釋放,增強組織抗炎功能,保護皮瓣內(nèi)微循環(huán)血流灌注,從而減輕皮瓣缺血再灌注損傷引起的病理生理變化,增強皮瓣活力,促進皮瓣存活。為雌二醇治療皮瓣缺血再灌注損傷提供了新的證據(jù)和方向。2.雌二醇可顯著抑制皮瓣組織中p38MAPK的蛋白表達,而增加MKP-2的蛋白表達,提示雌二醇可通過阻滯MAPK的炎癥信號級聯(lián)通路,調(diào)控細胞內(nèi)促炎性基因的表達,減輕皮瓣組織內(nèi)炎癥反應,該過程可能與MKP-2抑制p38MAPK的活性有關。
[Abstract]:Objective: p38 mitogen-activated protein kinase (p38MAPK) signaling pathway is one of the most important signaling pathways in mammalian cells to regulate inflammation, cell proliferation and apoptosis. It plays an important role in the occurrence and development of ischemia-reperfusion injury. The purpose of this study was to investigate the effects of estradiol (E 2) on the p38MAPK signal pathway in the flap and the role of the p38MAPK signal pathway in the ischemia reperfusion injury of the flap, and to provide a theoretical basis for the treatment of the flap ischemia reperfusion injury by estradiol. Methods Forty-eight male Wistar rats were used to establish the model of abdominal flap ischemia-reperfusion injury under the condition of no specific pathogenic bacteria (SPF grade) in the laboratory for 14 weeks. The rats were randomly divided into control group (group 鈪，

本文編號：2158035