高血糖调控脑源性神经营养因子前体表达加重脊髓缺血再灌注损伤模型小鼠神经损伤和炎症反应的机制  

Mechanism by which hyperglycemia regulates precursor of brain-derived neurotrophic factor expression to exacerbate neurological injury and inflammation in a mouse model of spinal cord ischemia-reperfusion injury

作　　者：罗伟[1] 缪雪梅 刘涛[1] 熊熠宇 戴茹萍[1] 李卉[1] LUO Wei;MIAO Xuemei;LIU Tao;XIONG Yiyu;DAI Ruping;LI Hui(Department of Anesthesiology,Second Xiangya Hospital,Central South University,Changsha 410011,China)

机构地区：[1]中南大学湘雅二医院麻醉科,长沙410011

出　　处：《中南大学学报(医学版)》2024年第12期1875-1884,共10页Journal of Central South University :Medical Science

基　　金：国家自然科学基金(81873770,82271430);湖南省自然科学基金(2023JJ30759)。

摘　　要：目的:目前脊髓缺血再灌注损伤(spinal cord ischemia-reperfusion injury,SCIRI)仍缺乏有效的防治手段,一直是脏器保护领域的难点和热点。高血糖可以通过多种机制导致神经损伤,是围术期常见的问题,但是对SCIRI的影响及机制尚不清楚。本研究探讨脑源性神经营养因子前体(precursor of brain-derived neurotrophic factor,proBDNF)在高血糖诱导SCIRI模型小鼠中的作用机制。方法:将8周龄雄性C57BL/6小鼠随机分为对照(Vehicle)组和糖尿病(diabetes mellitus,DM)组;DM组经腹腔注射链脲佐菌素(streptozotocin,STZ)联合饮用10%糖水构建DM模型小鼠,Vehicle组腹腔注射等体积50 mmol/L柠檬酸钠溶液(pH值为4.5);将小鼠空腹血糖水平≥11.1 mmol/L判定为DM模型构建成功。Vehicle组和DM组均通过夹闭降主动脉进行SCIRI造模,假手术(Sham)组只暴露降主动脉,不进行阻断。采用Basso小鼠运动功能评分量表(Basso Mouse Scale,BMS)及其子量表(sub-BMS)评估各组模型小鼠下肢运动功能,采用旷场实验评估模型小鼠自主活动能力。采用免疫组织化学染色观察脊髓神经核抗原(neuronal nuclearprotein,NeuN)和proBDNF的表达变化;采用实时反转录聚合酶链反应检测脊髓组织中白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的mRNA表达水平。观察proBDNF单克隆抗体(monoclonal anti-proBDNF antibody,McAb-proB)干预对DM模型小鼠SCIRI的影响,随机将DM模型小鼠进行分组,其中DM+SCIRI+McAb-proB组SCIRI造模前30 min经腹腔注射proBDNF单克隆抗体100μg,DM+SCIRI+Vehicle组经腹腔注射等量同类型免疫球蛋白G抗体;采用BMS和sub-BMS评估小鼠运动功能恢复情况,并检测脊髓组织中IL-1β、IL-6、TNF-α的mRNA表达水平。结果:与Vehicle+SCIRI组比较,DM+SCIRI组BMS、sub-BMS评分以及脊髓NeuN表达降低,运动总距离减少,运动速度降低,proBDNF表达水平以及IL-1β、IL-6�Objective:Spinal cord ischemia-reperfusion injury(SCIRI)remains a major challenge in the field of organ protection due to the lack of effective prevention and therapeutic strategies.Hyperglycemia,a common perioperative condition,contributes to neurological injury via multiple mechanisms.However,its role and underlying mechanism in SCIRI are still unclear.This study aims to investigate the involvement of the precursor of brain derived neurotrophic factor(proBDNF)in hyperglycemia-induced SCIRI in mice.Methods:Eight-week-old male C57BL/6 mice were randomly assigned to a control group(Vehicle)or a diabetes mellitus(DM)group.The DM group was established using intraperitoneal injection of streptozotocin(STZ)combined with 10%sucrose water.The Vehicle group received an equal volume of 50 mmol/L sodium citrate buffer(pH 4.5).Fasting blood-glucose levels≥11.1 mmol/L were considered successful DM modeling.Both Vehicle and DM groups underwent SCIRI modeling via descending aortic clamping,while the Sham group underwent a sham procedure without aortic occlusion.Lower limb motor function was assessed using the Basso Mouse Scale(BMS)and its subscale(sub BMS).Locomotor activity was evaluated using an open field test.Immunohistochemistry was performed to detect changes in neuronal nuclear protein(NeuN)and proBDNF expression in spinal cord tissues.Real-time reverse transcription polymerase chain reaction(RT-PCR)was used to measure mRNA expression of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α).To explore the effect of proBDNF inhibition,diabetic mice were divided into groups:A DM+SCIRI+monoclonal anti-proBDNF antibody(McAb-proB)group received an intraperitoneal injection of 100μg of McAb proB 30 minutes before SCIRI modeling,and a DM+SCIRI+Vehicle group received an equal amount of isotype immunoglobulin G.BMS and sub-BMS scores were recorded,and the gene expression of inflammatory cytokines mentioned above were evaluated.Results:Compared with the Vehicle+SCIRI group,the DM+SCIRI group showed s

关 键 词：糖尿病 脊髓缺血再灌注损伤 脑源性神经营养因子前体 神经炎症 高血糖状态 

分 类 号：R28[医药卫生—中药学]