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 es-tablished 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.Immunofluores-cence 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 alco-hol use disorder AUD group,alcohol use disorder stroke AUD+MCAO rats had elevated cerebral cortical TMEM 166 expression,obvious neurologi-cal deficits in the rats,and large cerebral infarcts on the ischemic side,and were found to be accompanied by decreased expression of Hint2 by im-munofluorescence and Western blotting detection.TMEM166 siRNA lateral ventricle injection was lowly expressed after TMEM166 injection,the fluorescence intensity of TMEM 166 protein was significantly weakened,while the fluorescence intensity of Hint2 protein was significantly en-hanced,the number of positive cells increased,the area of cerebral infarction in rats was significantly reduced,and the symptoms of neurological deficits improved.Meanwhile,TMEM 166 co-localized with Hint2.Conclusion TMEM 166 plays an important role in neurological deficits in al-cohol use disorder stroke in rats,and its mechanism may be realized by inhibiting the expression of Hint2.
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
Key words
Rats/Alcohol use disorders/Stroke/Transmembrane protein 166/Histidine trinucleotide binding protein 2
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