米诺环素对脑出血小鼠认知功能障碍的影响及其机制  

Effect of minocycline on cognitive dysfunction in mice with intracerebral hemorrhage and its mechanism

作　　者：李阳阳[1] 方建[2] 王晓雪[1] LI Yangyang;FANG Jian;WANG Xiaoxue(Department of Neurology for the Elderly,The First Affiliated Hospital of Henan University,Kaifeng 475001,China)

机构地区：[1]河南大学第一附属医院老年神经内科,河南开封475001 [2]河南大学第一附属医院神经内科二病区,河南开封475001

出　　处：《精准医学杂志》2025年第1期19-24,29,共7页Journal of Precision Medicine

基　　金：河南省医学科技攻关计划(LHGJ20210555)。

摘　　要：目的探讨米诺环素(Mcy)对脑出血(ICH)小鼠认知功能障碍的影响及其机制。方法将C57BL/6J雄性小鼠随机分为A~E组,每组20只。A组小鼠尾静脉注射生理盐水,B组小鼠尾静脉注射自体血构建ICH模型,C、D组小鼠在B组基础上每天分别灌胃20、50 mg/kg的Mcy,E组小鼠在D组基础上再腹腔注射5μL含有12μg MK-2206的DMSO溶液,共14 d。处理结束后依据改良神经病学严重程度评分(mNSS)评估各组小鼠神经损伤情况,采用水迷宫实验检测各组小鼠的认知能力,计算各组小鼠脑组织含水量,采用HE染色方法观察各组小鼠海马组织的病理形态,采用TUNEL法检测各组小鼠海马组织中神经细胞凋亡情况,采用酶联免疫吸附试验(ELISA)检测各组小鼠海马组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)水平,采用Western blot方法检测各组小鼠海马组织中Bax、Bcl-2、磷酸酶张力蛋白同源物(PTEN)、蛋白激酶B(Akt)及p-Akt蛋白水平。结果神经损伤评估结果显示,C、D组小鼠mNSS评分较B组显著降低(q=22.925、55.457,P<0.05),E组小鼠mNSS评分较D组显著升高(q=44.448,P<0.05);水迷宫实验结果显示,与B组比较,C、D组小鼠逃避潜伏期、探索时间延长(q=22.169~91.845,P<0.05),穿越次数增加(q=18.347、41.936,P<0.05);与D组比较,E组小鼠逃避潜伏期、探索时间延长(q=30.765、85.881,P<0.05),穿越次数减少(q=39.315,P<0.05);脑组织含水量和TUNEL染色检测结果显示,C、D组小鼠脑组织含水量、海马组织神经细胞凋亡率较B组降低(q=7.269~33.327,P<0.05),E组小鼠脑组织含水量、海马组织神经细胞凋亡率较D组升高(q=9.957、31.004,P<0.05);ELISA及Western blot实验检测结果显示,与B组比较,C、D组小鼠海马组织中MDA含量、Bax和PTEN相对表达量降低(q=10.734~22.978,P<0.05),SOD和GSH-Px水平、Bcl-2相对表达量及p-Akt/Akt值升高(q=11.862~31.997,P<0.05);与D组比较,E组小鼠海马组织中MDA含量、Bax和PObjective To investigate the effect of minocycline(Mcy)on cognitive dysfunction in mice with intracerebral hemorrhage(ICH)and its mechanism.Methods Male C57BL/6J mice were randomly divided into groups A,B,C,D,and E,with 20 mice in each group.The mice in group A were given injection of normal saline via the caudal vein,those in group B were given injection of autologous blood via the caudal vein to establish a model of ICH,those in groups C and D were given Mcy by gavage every day at a dose of 20 mg/kg and 50 mg/kg,respectively,in addition to the treatment in group B,and those in group E were given intraperitoneal injection of 5μL DMSO solution containing 12μg MK-2206 in addition to the treatment in group D;the course of treatment was 14 days for all groups.After treatment,modified Neurological Severity Score(mNSS)was used to assess nerve injury of mice in each group;the water maze test was used to evaluate the cognitive ability of mice in each group;brain water content was calculated for mice in each group;HE staining was used to observe the pathological morphology of hippocampal tissue;the TUNEL method was used to measure neuronal apoptosis in hippocampal tissue of mice in each group;ELISA was used to measure the levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)in hippocampal tissue of mice,and Western blot was used to measure the protein expression levels of Bax,Bcl-2,phosphatase and tensin ho-molog(PTEN),protein kinase B(Akt),and phosphorylated Akt(p-Akt)in hippocampal tissue.Results Nerve injury assessment showed that groups C and D had a significantly lower mNSS score than group B(q=22.925,55.457,P<0.05),and group E had a significantly higher mNSS score than group D(q=44.448,P<0.05).The water maze test showed that compared with group B,groups C and D had significantly longer escape latency and exploration time(q=22.169-91.845,P<0.05)and a significantly higher number of crossings(q=18.347,41.936,P<0.05),and compared with group D,group E had significantly longer esca

关 键 词：米诺环素 脑出血 PTEN磷酸水解酶 原癌基因蛋白质c-akt 信号传导 认知障碍 疾病模型 动物 

分 类 号：R743.34[医药卫生—神经病学与精神病学] R741[医药卫生—临床医学]