饮水型砷暴露引起广泛性焦虑障碍及血浆外泌体特异性miRNAs研究  

Generalized anxiety disorder and plasma exosome-specific miRNAsinduced by drinking-water arsenic exposure

作　　者：尹紫月 郭林楠 雷炜星 郭雪峰 崔钧贺 余启明 张磊 王娜 YIN Zi-yue;GUO Lin-nan;LEI Wei-xing;GUO Xue-feng;CUI Jun-he;YU Qi-ming;ZHANG Lei;WANG Na(School of Public Health,Guilin Medical University,Guilin,Guangxi 541000,China;不详)

机构地区：[1]桂林医学院公共卫生学院,广西桂林541000 [2]东北大学生命科学与健康学院

出　　处：《现代预防医学》2025年第4期622-628,共7页Modern Preventive Medicine

基　　金：国家自然科学基金(82160642);广西自然科学基金(2021GXNSFBA196014,2024JJA140974)。

摘　　要：目的了解饮水型砷暴露致广泛性焦虑障碍(Generalizedanxietydisorder,GAD)的作用,探索血浆外泌体miRNAs的特异性改变。方法将16只SPF级wistar雄性大鼠随机分到正常组和染砷组,每组8只,染砷组饮用水中的NaAsO_(2)浓度0.10 mg/L,两组均可自由饮水。共染毒21天,分别用旷场和高架十字迷宫实验检测染毒的第0、7、14、21天的行为学,以评估GAD样行为;实验结束后,提取两组大鼠的血浆外泌体miRNAs进行测序,并运用实时荧光定量PCR验证差异miRNAs的表达情况。结果染毒后的第21 d染砷组的体重低于正常组(t=7.950,P<0.001)。旷场实验结果显示,与正常组相比,染砷组的总路程减少(t=2.213,P=0.044),修饰次数增加(t=-4.704,P<0.001)、跨格次数(t=4.340,P=0.001)和站立次数减少(t=4.496,P=0.001);高架十字迷宫实验结果显示,与正常组相比,染砷组的开臂次数(t=3.614,P=0.003)和开臂时间(t=4.775,P<0.001)减少,闭臂次数(t=-2.486,P=0.026)与闭臂时间增加(t=-6.862,P<0.001),砷暴露引起了大鼠GAD样行为。实时荧光定量PCR结果显示,与正常组相比,染砷组中的miR-99b-3p、miR-9a-5p、miR-218a-5p表达均上调;miR-425a-3p、miR-378a-3p、miR-155-5p表达均下调(P<0.05)。GO、KEGG基因富集分析结果显示,差异血浆外泌体miRNAs可能通过内吞作用、MAPK通路、Ras信号通路、cAMP信号通路、mTOR信号通路、凋亡来参与GAD的发生发展。结论饮水型砷暴露可以引起大鼠的GAD样行为,外泌体miR-99b-3p、miR-9a-5p、miR-218a-5p、miR-425a-3p、miR-378a-3p、miR-155-5p可能参与调节饮水型砷暴露致GAD的过程。Objective To examine the impact of drinking water arsenic exposure on the development of generalized anxiety disorder(GAD)and to investigate specific changes in plasma exosomal miRNAs.Methods Sixteen male SPF-grade Wistar rats were randomly divided into two groups:control and arsenic.Each group consisted of 8 rats.The arsenic group was exposed to 0.10 mg/L.All groups had unrestricted access to drinking water.The rats were exposed to arsenic for a duration of 21 days.Behavioral assessments using the open field and elevated plus maze tests were conducted on days 0,7,14,and 21 to evaluate GAD-like behavior.At the end of the experiment,plasma exosomal miRNAs were extracted from both groups of rats.Sequencing was performed,and the expression levels of differentially expressed miRNAs were validated using real-time quantitative PCR.Results On the 21st day after arsenic exposure,the arsenic group exhibited lower body weight compared to the control group(t=7.950,P<0.001).In the open field test,the arsenic group showed a decrease in total distance traveled(t=2.213,P=0.044),an increase in rearing frequency(t=-4.704,P<0.001),an increase in grid crossings(t=4.340,P=0.001),and a decrease in standing frequency(t=4.496,P=0.001)compared to the control group.In the elevated plus maze test,the arsenic group exhibited a decrease in the number of entries into the open arms(t=3.614,P=0.003)and a decrease in time spent in the open arms(t=4.775,P<0.001),while showing an increase in the number of entries into the closed arms(t=-2.486,P=0.026)and an increase in time spent in the closed arms(t=-6.862,P<0.001)compared to the control group.These results indicate GAD-like behavior in the rats exposed to arsenic.Real-time quantitative PCR revealed that the expression levels of miR-99b-3p,miR-9a-5p,and miR-218a-5p were upregulated,while the expression levels of miR-425a-3p,miR-378a-3p,and miR-155-5p were downregulated in the arsenic group compared to the control group(P<0.05).Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enr

关 键 词：饮水型砷暴露 广泛性焦虑障碍 血浆外泌体miRNAs 

分 类 号：R-33[医药卫生] R123