db/db小鼠远端大脑中动脉闭塞模型行为学评估方法的筛选  

Neurofunctional assessment of db/db mice in permanent distal middle cerebral artery occlusion

作　　者：马舒贝[1] 孙威[2] 赵海苹 罗玉敏 Ma Shubei;Sun Wei;Zhao Haiping;Luo Yumin(Department of Neurology,Dalian Municipal Central Hospital,Dalian,Liaoning 116033,China;不详)

机构地区：[1]辽宁省大连市中心医院神经内科,116033 [2]大连市第三人民医院神经内科 [3]首都医科大学宣武医院脑血管病研究室

出　　处：《中国脑血管病杂志》2020年第9期520-530,共11页Chinese Journal of Cerebrovascular Diseases

基　　金：国家自然科学基金资助项目(81771413);辽宁省自然科学基金指导计划项目(2019-ZD-0884)。

摘　　要：目的通过对比分析瘦素受体突变导致自发性2型糖尿病的db/db小鼠远端大脑中动脉闭塞(dMCAO)后行为学评估结果,筛选适合db/db小鼠dMCAO模型的行为学评估方法。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野生型小鼠(WT)作为双对照组。采用dMCAO作为脑缺血模型。测量各品系小鼠血糖水平及dMCAO后3、35 d时脑组织缺损体积以评估合并糖尿病的卒中模型造模是否成功。选择以下行为学评估方法进行分析:转棒测试、圆筒测试、粘贴撕除测试、错步测试方法评估dMCAO模型的感觉运动功能及运动协调性;选择Morris水迷宫测试和新事物认知测试评估模型认知功能;选择旷场测试评估模型焦虑状态和一般运动能力及探索能力。结果db/db小鼠dMCAO前后血糖水平均明显高于db/+和WT小鼠(均P<0.01);dMCAO术后3 d,db/db小鼠脑组织缺损体积明显大于db/+小鼠[(19.3±1.0)%比(10.9±1.5)%,P<0.01]和WT小鼠[(12.1±1.4)%,P<0.05],证明合并糖尿病的卒中模型造模成功。行为学评估结果:(1)粘贴撕除测试。假手术组db/db、db/+和WT小鼠间的接触粘纸时间(品系效应:F=2.56,P=0.08)和移除粘纸时间(品系效应:F=0.18,P=0.84)差异均无统计学意义。dMCAO组术后db/db小鼠与db/+、WT小鼠间接触粘纸时间(品系效应:F=10.94,P<0.01)和移除粘纸时间(品系效应:F=22.20,P<0.01)差异均有统计学意义,db/db小鼠接触粘纸时间和移除粘纸时间均明显多于db/+、WT小鼠(P<0.05或P<0.01)。(2)错步测试。假手术组和dMCAO组中,db/db、db/+和WT小鼠间错步率差异均无统计学意义(品系效应:F=0.84,P=0.48;F=3.50,P=0.06)。db/db、db/+和WT小鼠dMCAO后第1天和第3天的平均错步率与dMCAO术前比较,明显上升(均P<0.05),dMCAO术后第28天和第35天平均错步率均较dMCAO术前下降(均P<0.05)。(3)新物体认知测试。假手术组和dMCAO组中db/db、db/+、WT小鼠间探索新物体的次数差异有统�Objective By comparing and analyzing the behavioral testing results after distal middle cerebral artery occlusion(dMCAO)in db/db mice with spontaneous type 2 diabetes mellitus due to leptin receptor mutation,the neurofunctional assessment methods suitable for dMCAO model in db/db mice were selected.Methods We used db/db mice to establish a type 2 diabetes mellitus model.At the same time,we chose heterozygous db/+mice and C57 wide type(WT)mice as double control groups.Permanent distal middle cerebral artery occlusion was performed as an ischemic stroke model.Blood glucose level and brain-tissue loss volume were measured at day 3 and day 35 after dMCAO to evaluate the success of the diabetic stroke model.The following behavioral tests,including the rotarod,cylinder,adhesive removal,and foot fault tests were selected to evaluate the sensorimotor function and motor coordination in dMCAO model.The Morris water maze test and novel object recognition test were conducted to evaluate cognitive function.The open-field test was performed to estimate anxiety-like behavior,general motor and exploring ability in mice.Results Blood glucose levels in db/db mice before and after dMCAO were significantly higher than that in db/+mice and WT mice(all P<0.01).Three days after dMCAO operation,the brain-tissue loss volume of db/db mice was significantly larger than that of db/+mice([19.3±1.0]%vs.[10.9±1.5]%,P<0.01)and WT mice([12.1±1.4]%,P<0.05),indicating that the diabetic stroke model was successfully modeled.Behavioral testing results:(1)Adhesive removal test.In sham operation group,there were no statistically significant differences among db/db,db/+and WT mice in contact time(strain effect:F=2.56,P=0.08)and removal time(strain effect:F=0.18,P=0.84).In dMCAO group,there were statistically significant differences among db/db,db/+and WT mice in contact time(strain effect:F=10.94,P<0.01)and removal time(strain effect:F=22.20,P<0.01).The contact time and removal time in db/db mice were significantly higher than that in db/+mice and W

关 键 词：DB/DB小鼠 行为学评估 远端大脑中动脉闭塞 

分 类 号：R743.3[医药卫生—神经病学与精神病学]