TMEM166在大鼠酒精使用障碍脑卒中中的作用及其与Hint2表达的关系  

Role of TMEM166 in alcohol use disorder stroke in rats and its relationship with Hint2 expression

作　　者：郝丹丹 王丹阳 李丽[1] HAO Dan-dan;WANG Dan-yang;LI Li(Department of Anesthesiology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院麻醉科,北京100050

出　　处：《临床和实验医学杂志》2024年第15期1569-1572,共4页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金面上项目(编号:82271307)。

摘　　要：目的探讨跨膜蛋白166(TMEM166)在大鼠酒精使用障碍(AUD)脑卒中中的作用及其与组氨酸三核苷酸结合蛋白2(Hint2)表达的关系。方法实验动物为SPF级成年SD雄性大鼠54只,实验中将大鼠按照随机数字表法分为3组:AUD组、AUD+大脑中动脉栓塞(MCAO)组及AUD+MCAO+TMEM166siRNA(TMEM166 siRNA)组,每组各18只。AUD组用双瓶法建立AUD动物模型,AUD+MCAO组在AUD基础上制作大鼠MCAO局部脑缺血模型,AUD+MCAO+TMEM166siRNA在AUD+MCAO大鼠基础上侧脑室注射TMEM166 siRNA。记录各组大鼠乙醇摄入量、乙醇偏好变化及神经功能评分。免疫荧光染色观察各组大鼠大脑皮层TMEM166与Hint2的表达,蛋白质印迹法检测各组大鼠脑组织TMEM166表达及Hint2的蛋白水平。结果与AUD组相比,AUD+MCAO组大鼠大脑皮质TMEM166表达升高,大鼠神经功能缺损明显,缺血侧出现大面积脑梗死,免疫荧光及蛋白质印迹法检测发现伴有Hint2的表达降低。TMEM166 siRNA侧脑室注射低表达TMEM166后,TMEM166蛋白荧光强度明显减弱,而Hint2蛋白荧光强度明显增强,阳性细胞数量增加,大鼠脑梗死面积明显减少,神经功能缺损症状改善。同时,TMEM166与Hint2存在共定位。结论TMEM166在大鼠酒精使用障碍脑卒中神经功能障碍中发挥重要作用,其机制可能是通过抑制Hint2的表达实现的。Objective To investigate the role of transmembrane protein 166(TMEM166)in alcohol use disorder(AUD)stroke in rats and its relationship with the expression of histidine trinucleotide binding protein 2(Hint2).Methods The experimental animals were 54 adult SD male rats of SPF grade.According to the random number table method,the rats were divided into three groups:the AUD group,the AUD+middle cerebral artery embolism(MCAO)group and the AUD+MCAO+TMEM166siRNA group,with 18 rats in each group.The AUD group established the AUD animal model using the two-bottle method,the AUD+MCAO group was made local cerebral ischemia model of MCAO on the basis of AUD,and the AUD+MCAO+TMEM166siRNA group were injected into lateral ventricles with TMEM166 siRNA on the basis of AUD+MCAO group.Ethanol intake,ethanol preference changes and neurological function scores of rats in each group were recorded.Immunofluorescence staining was used to observe the expression of TMEM166 and Hint2 in the cerebral cortex of the animals in each group,and Western blotting detected the expression of TMEM166 and the protein level of Hint2 in the brain tissue of the rats in each group.Results Compared with the alcohol use disorder AUD group,alcohol use disorder stroke AUD+MCAO rats had elevated cerebral cortical TMEM166 expression,obvious neurological deficits in the rats,and large cerebral infarcts on the ischemic side,and were found to be accompanied by decreased expression of Hint2 by immunofluorescence and Western blotting detection.TMEM166 siRNA lateral ventricle injection was lowly expressed after TMEM166 injection,the fluorescence intensity of TMEM166 protein was significantly weakened,while the fluorescence intensity of Hint2 protein was significantly enhanced,the number of positive cells increased,the area of cerebral infarction in rats was significantly reduced,and the symptoms of neurological deficits improved.Meanwhile,TMEM166 co-localized with Hint2.Conclusion TMEM166 plays an important role in neurological deficits in alcohol use disorder stroke

关 键 词：大鼠 酒精使用障碍 脑卒中 跨膜蛋白166 组氨酸三核苷酸结合蛋白2 

分 类 号：R749.62[医药卫生—神经病学与精神病学] R743.3[医药卫生—临床医学]