脑白质损伤加重db/db小鼠远端大脑中动脉闭塞后焦虑行为  

White matter injury exacerbated anxiety-like behavior of db/db mice after distal middle cerebral artery occlusion

作　　者：马舒贝[1] 王建懿 孙威[3] 潘心[1] 赵红玲[1] Ma Shubei;Wang Jianyi;Sun Wei;Pan Xin;Zhao Hongling(Department of Neurology,Dalian Municipal Central Hospital,Dalian 116033,China;Department of Neuology,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China;Department of Neurology,Dalian No.3 People's Hospital,Dalian 116033,China)

机构地区：[1]大连市中心医院神经内科,大连116033 [2]西安交通大学第一附属医院神经内科,西安710061 [3]大连市第三人民医院神经内科,大连116033

出　　处：《中国医师进修杂志》2022年第5期407-414,共8页Chinese Journal of Postgraduates of Medicine

基　　金：辽宁省自然科学基金指导项目(2019-ZD-0884);大连市医学科学研究计划项目(1911005)。

摘　　要：目的研究db/db小鼠远端大脑中动脉闭塞(dMCAO)后焦虑行为加重的机制。方法选择db/db小鼠作为2型糖尿病模型,选择同窝杂合子小鼠(db/+)和C57野生型(WT)小鼠作为双对照组。采用dMCAO模型作为急性脑缺血模型。测量假手术组和dMCAO前后各品系小鼠的血糖水平。通过免疫荧光染色检测微管相关蛋白-2表达,评估dMCAO后35 d时各品系小鼠的脑组织缺损体积。选择旷场测试评估小鼠的焦虑行为。选择免疫荧光染色检测非磷酸化神经丝蛋白4(SMI32)和髓鞘碱性蛋白(MBP)的表达以评估小鼠dMCAO术后35 d时脑白质损伤情况,并将SMI32和MBP的免疫荧光强度比值与旷场测试的中心停留时间和角落停留时间进行相关性分析以评估脑白质损伤程度与焦虑行为的相关性。选择流式细胞技术检测小鼠缺血侧脑组织中T细胞,包括调节性T细胞(Tregs)和B细胞的数量。结果假手术组和dMCAO组中db/db小鼠术前和术后血糖水平明显高于db/+小鼠和WT小鼠(均P<0.01)。db/db小鼠dMCAO后35 d时3个品系小鼠脑组织缺损体积比较差异无统计学意义(P>0.05)。旷场测试结果:假手术组和dMCAO组db/db小鼠在旷场行走总路径距离均明显少于db/+小鼠、WT小鼠(P<0.05或<0.01)。db/+小鼠在旷场行走总路径距离少于WT小鼠(P<0.05)。假手术组db/db小鼠在旷场中心停留时间明显少于db/+小鼠、WT小鼠(P<0.05或<0.01)。dMCAO组db/db小鼠在旷场中心停留时间明显少于db/+、WT小鼠(P<0.05或<0.01),db/+小鼠少于WT小鼠(P<0.01)。假手术组db/db、db/+小鼠在旷场角落停留时间明显多于WT小鼠(db/db小鼠比WT小鼠,P<0.01;db/+小鼠比WT小鼠,P<0.05)。dMCAO组db/db小鼠在旷场角落停留时间明显多于db/+、WT小鼠(P<0.05或<0.01),db/+小鼠多于WT小鼠(P<0.05)。免疫荧光染色检测小鼠脑梗死周边皮层和外囊区SMI32和MBP表达结果。dMCAO组db/db小鼠脑梗死周边外囊区SMI32/MBP荧光强度比值明显高于db/+小鼠Objective To explore the possible mechanism of exacerbation of anxiety-like behavior in db/db mice after distal middle cerebral artery occlusion(dMCAO).Methods The db/db mice was used to establish a type 2 diabetes mellitus model.Meanwhile,heterozygous db/+mice and C57 wild-type(WT)mice were chosen as double control groups.Then a permanent distal middle cerebral artery occlusion model was employed as an acute ischemic stroke model.The blood glucose levels before and post-dMCAO surgery on day1,day3,and day5 were detected.The brain tissue loss at 35 days after stroke was measured by immunofluorescent staining of MAP2.The open-field test was performed to estimate anxiety-like behavior and general motor and exploring ability of the animals.Axons and myelin were immunostained with non-phosphorylated neurofilaments(SMI32)and myelin basic protein(MBP),respectively,to evaluate differences in white matter integrity in WT,db/+and db/db mice 35 days after stroke.The correlation between SMI32/MBP and open field test parameters(time in center and corner)was analyzed.Flow cytometry was employed to detect the amount of T cells and B cells,including regulatory T cells(Tregs)in the brain tissue.Results Blood glucose levels in db/db mice were significantly higher than db/+mice and WT mice in both sham and dMCAO groups(P<0.01).There was no significant difference in brain tissue loss 35 days post-stroke among db/db mice,db/+mice,and WT mice.In the open field test,there were significant differences in the total distance of db/db mice,db/+and WT mice in the sham and dMCAO groups.Db/db mice shorter than db/+mice(P<0.05),WT mice(P<0.01),and db/+mice shorter than WT mice(P<0.05).There were significant time differences in the center among db/db,db/+,and WT mice in sham and dMCAO groups.In both the sham and dMCAO groups,db/db mice spent less time in the center area of the open field than WT mice(P<0.01).In the sham group,db/+mice spent less time in the center area than WT mice(P<0.05).In dMCAO group,db/db mice spent less time in the center

关 键 词：小鼠 焦虑 远端大脑中动脉闭塞 脑白质损伤 

分 类 号：R743.3[医药卫生—神经病学与精神病学]