miR-206靶向BDNF对七氟醚麻醉后诱导小鼠认知功能障碍的影响  

The effect of miR-206 targeting BDNF on cognitive impairment in mice induced by sevoflurane anesthesia

作　　者：丁玉美[1] 燕兴梅[1] 王彩霞[1] 武利栓[1] 邱颐[1] Ding Yumei;Yan Xingmei;Wang Caixia;Wu lishuan;Qiu Yi(Department of Anesthesiology,the Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,China)

机构地区：[1]内蒙古医科大学第二附属医院麻醉科,呼和浩特010030

出　　处：《脑与神经疾病杂志》2021年第12期737-741,共5页Journal of Brain and Nervous Diseases

基　　金：国家自然科学基金(81560192)。

摘　　要：目的探讨微小RNA-206 (microRNA-206,miR-206)靶向增强脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)对七氟醚麻醉后诱导小鼠认知功能障碍的影响。方法选择清洁级SD雄性小鼠32只,将小鼠随机分为4组:A组,对照组;B组,七氟醚组;C组,七氟醚+NC antagomir组;D组,七氟醚+miR-206 antagomir组。每组8只。采用Morris水迷宫测试测定小鼠认知功能;苏木精-伊红染色观察各组小鼠脑海马组织病理形态学改变;qRT-PCR检测各组小鼠脑组织miR-206及BDNF mRNA表达水平;Western blot检测各组小鼠脑组织BDNF蛋白表达水平;双荧光素酶验证miR-206与BDNF基因的靶向关系;流式细胞术检测脑海马组织神经元凋亡水平及凋亡基因Bax和Bcl-2的蛋白及mRNA表达水平。结果与A组相比,B组和C组小鼠逃避潜伏期长,平台所在象限停留时间及穿越平台次数减少(P<0.05);与C组相比,D组小鼠上述指标均有所改善(均P<0.05)。B组和C组小鼠脑组织miR-206的表达水平、脑海马组织神经元凋亡率及Bax表达水平与A组相比均升高,而BDNF及Bcl-2表达水平与A组相比均降低(;P<0.05);抑制miR-206表达后,小鼠脑海马组织神经元凋亡率及Bax蛋白及mRNA表达水平均降低,而脑组织BDNF和Bcl-2蛋白和mRNA表达水平与B组和C组相比均升高(均P<0.05)。BDNF是miR-206 的靶基因。结论七氟醚麻醉后诱发小鼠认知功能障碍与脑组织神经元凋亡有关,抑制 miRNA-206 的表达可通过促进BDNF的表达水平改善认知功能损害。Objective To investigate the effect of microRNA-206(miR-206) targeting brain-derived neurotrophic factor(BDNF) on cognitive impairment in mice after sevoflurane anesthesia.Methods Thirty two SD male clean grade mice were randomly divided into four groups:group A:control group;group B:sevoflurane group;group C:Sevoflurane+NC antigomir group;group D:Sevoflurane+miR-206 antigomir group.8 in each group.Using Morris water maze test to measure the cognitive function of mice.HE staining was used to observe the pathomorphological changes of hippocampal tissue of mice in each group.The expression of miR-206 and BDNF mRNA was detected by qRT-PCR and BDNF protein was detected by Western blot.Double luciferase assay for targeting relationship between miR-206 and BDNF gene.Flow cytometry was used to detect the level of neuron apoptosis and the expression of Bax and Bcl-2.Results Compared with group A,mice in group B and group C had a longer escape latency,and the time of staying in the quadrant where the platform was and the times of crossing the platform were reduced(P <0.05);compared with group C,the above indexes in group D were improved(P <0.05).The expression level of miR-206,apoptosis rate of neurons,Bax protein and mRNA in brain hippocampus of group B and group C were increased than that in group A,while the expression level of BDNF,Bcl-2 protein and mRNA were decreased than that in group A(;P <0.05).After inhibiting the expression of miR-206,the apoptosis rate and the expression level of Bax in the hippocampus decreased,while the expression level of BDNF and Bcl-2 in the brain increased compared with group B and group C(;P <0.05).BDNF was the target gene of miR-206.Conclusion The cognitive dysfunction induced by sevoflurane anesthesia is related to the apoptosis of neurons in brain tissue.Inhibition of miR-206 expression can improve the cognitive impairment by promoting the expression level of BDNF.

关 键 词：微小RNA-206 增强脑源性神经营养因子 七氟醚 认知功能障碍 神经元凋亡 

分 类 号：R749.1[医药卫生—神经病学与精神病学]