性别及雌激素对脓毒症时细胞信号转导通路活化的影响

发布时间：2018-06-12 15:14

本文选题：脓毒症 + Toll样受体4　；参考：《中国人民解放军军医进修学院》2005年博士论文

【摘要】：目的:1.研究脓毒症状态下体内重要脏器TLR4/NF-κB信号传导通路的活化规律及其对细胞因子的产生和脏器损害的影响。2.研究性别差异及外源性雌激素对脓毒症时重要脏器组织TLR4/NF-κB信号通路活化调节的影响,探讨将雌激素用于预防和治疗脓毒症的可行性。方法:采用内毒素(5mg/kg)攻击大鼠为脓毒症模型。首先采用雄性Wistar大鼠100只,随机分3组:(1)正常对照组(M)(n=20),麻醉后活杀动物;(2)脓毒症组(ML)(n=60),首先复制脓毒症模型,然后分别于伤后1h、2h、6h麻醉状态下活杀动物,每个时间点各20只动物。(3)雌激素+脓毒症组(MEL)(n=20),首先给予苯甲酸雌二醇0.5mg/kg皮下注射,连续5d,然后复制脓毒症模型,于伤后2h麻醉状态下活杀动物。再选用雌性间情期Wistar大鼠80只随机分4组:(1)正常对照组(F)(n=20),麻醉后活杀动物;(2)脓毒症组(FL)(n=20),复制脓毒症模型,于伤后2h麻醉状态下活杀动物;(3)去卵巢脓毒症组(FOL)(n=20),先行双侧卵巢切除术,术后30d复制脓毒症模型,于伤后2h麻醉状态下活杀动物;(4)去卵巢+雌激素脓毒症组(FOEL)(n=20),先行去卵巢术,术后30d给予苯甲酸雌二醇0.5mg/kg皮下注射,连续5d,然后复制脓毒症模型并于伤后2h麻醉状态下活杀动物。各组大鼠首先常规消毒腹部皮肤,无菌打开腹腔,分离腹主动脉,经腹主动脉采血,离心分离血清标本-70℃冻存。无菌留取动物心肌、肝脏、肺脏组织置于-70℃冻存。采用逆转录-聚合酶链反应技术(RT-PCR)检测各脏器组织TLR4、MD-2及TNF-α mRNA的表达;采用免疫印迹法(Western blot)检测各脏器组织细胞膜蛋白中TLR4、MD-2蛋白表达情况;采用凝胶电泳迁移率改变分析法(EMSA)分析各脏器组织核因子-κB(NF-κB)活性;采用酶联免疫吸附试验(ELISA)检测血清中TNF-α含量的变化;采用免疫组化方法检测各组织TLR4、MD-2的表达强度;采用放射免疫法检测血清雌二醇浓度。结果:1.雄性脓毒症大鼠组织TLR4/NF-κB信号传导通路的活化规律。(1)正常对照组大鼠心肌、肝脏、肺脏均可表达一定量TLR4、MD-2 mRNA,伤
[Abstract]:Objective : 1 . To investigate the activation of the signaling pathway of Toll - like receptor 4 / NF - 魏B in sepsis and its effects on cytokine production and organ damage . 2 . To study the effects of sex difference and exogenous estrogen on the activation and regulation of the signaling pathway of the important organ in sepsis , and to explore the feasibility of using estrogen in the prevention and treatment of sepsis .

Methods : 100 male Wistar rats were randomly divided into three groups : ( 1 ) normal control group ( M ) ( n = 20 ) , after anesthesia , and ( 2 ) sepsis group ( ML ) ( n = 60 ) . ( 3 ) The rats were killed by subcutaneous injection of estradiol benzoate 0.5 mg / kg for 5 days , then the sepsis model was replicated in 2 hours after injury ; ( 2 ) the serum samples were isolated by reverse transcription - polymerase chain reaction ( EMSA ) .

Results : 1 . The activation of the signal transduction pathway was observed in the rat tissue of male sepsis . ( 1 ) The myocardial , liver and lungs of the normal control group were able to express a certain amount of TLR , MD - 2 mRNA and injury .
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R363

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