腺相关病毒介导的高迁移率族蛋白1抗体基因转导治疗小鼠脓毒症的效果  

Effectiveness of adeno-associated virus-mediated gene transduction of high mobility group box-1 protein antibody for treatment of sepsis in mice

作　　者：孙立柱 张晟 马泽通 林兆奋[1] 胡适 SUN Li-zhu;ZHANG Sheng;MA Ze-tong;LIN Zhao-fen;HU Shi(Department of Emergency Medicine and Critical Medicine,Changzheng Hospital,Naval Medical University(Second Military Medical University),Shanghai 200003,China;Department of Critical Care Medicine,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China;Department of Biophysics,College of Basic Medical Sciences,Naval Medical University(Second Military Medical University),Shanghai 200433,China)

机构地区：[1]海军军医大学(第二军医大学)长征医院急救科,上海200433 [2]上海交通大学医学院附属瑞金医院重症医学科,上海200025 [3]海军军医大学(第二军医大学)基础医学院生物物理教研室,上海200433

出　　处：《第二军医大学学报》2021年第3期249-254,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(31970882)。

摘　　要：目的探讨腺相关病毒(AAV)介导的高迁移率族蛋白1(HMGB1)抗体基因转导对脓毒症的治疗效果。方法60只雌性C57小鼠随机平均分为2批,每批再分为AAV-HMGB1抗体组、AAV-对照-IgG组、生理盐水组,每组10只。AAV-HMGB1抗体组小鼠左侧股直肌肌内注射60μL AAV-HMGB1抗体病毒,AAV-对照-IgG组左侧股直肌肌内注射60μL AAV-对照-IgG病毒,生理盐水组左侧股直肌肌内注射60μL生理盐水,4周后对所有小鼠进行盲肠结扎穿孔手术制造脓毒症模型。第1批小鼠用于观察术后14 d生存率。第2批小鼠术后通过内眦静脉取血,检测血清炎症因子IL-6、IL-1β、TNF-α水平,取心、肝、肺、肾、小肠组织制作病理切片。结果使用AAV-HMGB1抗体基因转导产生HMGB1抗体对小鼠有抗脓毒症保护效应,90%(9/10)的脓毒症小鼠存活到造模后第14天;而注射生理盐水和AAV-对照-IgG病毒的小鼠在造模后第14天存活者仅分别占60%(6/10)和50%(5/10)。相比生理盐水组和AAV-对照-IgG组,AAV-HMGB1抗体组小鼠血清炎症因子IL-6、IL-1β、TNF-α水平降低(P均<0.05)。使用AAV-HMGB1抗体基因转导产生的HMGB1抗体对脓毒症小鼠的肺组织有明显保护作用,但对其他脏器的保护作用不明显。结论AAV介导的HMGB1抗体基因转导可通过高表达HMGB1抗体中和血清中的HMGB1,从而降低脓毒症小鼠的炎症因子水平,改善重要器官病理变化,并可能降低脓毒症小鼠死亡率,有望成为治疗脓毒症的新策略。Objective To investigate the therapeutic effect of adeno-associated virus(AAV)-mediated gene transduction of high mobility group box-1 protein(HMGB1)antibody on sepsis.Methods Sixty female C57 mice were evenly randomized into 2 sets,and each set was divided into 3 groups(n=10):AAV-HMGB1 antibody group,AAV-controlIg G group,and normal saline group,and the mice were injected intramuscularly with 60μL of AAV-HMGB1 antibody virus,60μL of AAV-control-Ig G virus,and 60μL of normal saline in the left rectus femoris muscle in the 3 groups,respectively.All mice were subjected to cecal ligation and perforation 4 weeks later to establish sepsis models.The 14-d survival rate of mice in the first set was observed.Serum levels of inflammatory factors(interleukin[IL]-6,IL-1βand tumor necrosis factorα[TNF-α])were measured in the blood obtained from the angular vein of mice in the second set,and pathological sections were taken from the heart,liver,lung,kidney and small intestine tissues.Results The HMGB1 antibody produced by the AAV-HMGB1 antibody gene transduction had a protective effect against sepsis in mice,90%(9/10)of septic mice survived until day 14 post-modeling in the AAV-HMGB1 antibody group,while only 60%(6/10)and 50%(5/10)of mice survived on day 14 post-modeling in the normal saline group and the AAV-control-Ig G group,respectively.The serum levels of inflammatory factors(IL-6,IL-1βand TNF-α)were significantly lower in the AAV-HMGB1 antibody group compared with those in the normal saline and AAV-control-Ig G groups(all P<0.05).The AAV-HMGB1 antibody produced by the AAV-HMGB1 antibody gene transduction had a significant protective effect on the lung tissue,but not on other organs.Conclusion AAV-mediated gene transduction of HMGB1 antibody can neutralize HMGB1 in the serum by high expression of HMGB1 antibody,thus reducing the level of inflammatory factors in septic mice,improving pathological changes in vital organs and possibly reducing mortality in septic mice,which is expected to be a new strategy for the t

关 键 词：高迁移率族蛋白1 脓毒症 炎症因子 基因转导 抗体治疗 腺相关病毒 

分 类 号：R631[医药卫生—外科学]