早期游泳调节纹状体细胞自噬缓解Shank3基因敲除大鼠刻板行为  

Early Swimming Alleviates Stereotypic Behavior in Shank3 Knockout Rats by Regulating Striatal Cell Autophagy

作　　者：薛亚奇 刘纽 王世娇 把懿 甄志平[1] XUE Ya-Qi;LIU Niu;WANG Shi-Jiao;BA Yi;ZHEN Zhi-Ping(College of P.E and Sports,Beijing Normal University,Beijing 100875,China;College of P.E,Weinan Normal University,Weinan 714099,China)

机构地区：[1]北京师范大学体育与运动学院,北京100875 [2]渭南师范学院体育学院,渭南714099

出　　处：《生物化学与生物物理进展》2025年第3期751-763,共13页Progress In Biochemistry and Biophysics

基　　金：北京市自然科学基金(7232239)资助项目。

摘　　要：目的通过8周游泳运动干预Shank3基因敲除(Shank3^(-/-))诱导的孤独症谱系障碍(autism spectrum disorder,ASD)大鼠模型,基于细胞自噬视角探索运动干预改善大鼠孤独症样行为的机制。方法根据基因型鉴定结果及运动干预情况将大鼠分为野生对照组(WC组)、Shank3^(-/-)组(KC组)、野生游泳组(WS组)、Shank3^(-/-)游泳组(KS组),每组15只。KS组和WS组进行8周游泳运动锻炼,5 d/周,逐步递增至40 min/次并保持。游泳运动干预后24 h进行行为学实验,包括:自梳理实验、埋珠实验、孔洞实验。行为学测试12 h后进行取材,通过透射电镜观察纹状体区域自噬体数量,免疫荧光染色观察微管相关蛋白1轻链3(LC3)蛋白及选择性自噬接头蛋白(p62)蛋白表达水平,实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qPCR)和免疫印迹法(Western blot)检测纹状体组织中B细胞淋巴瘤蛋白2相互作用蛋白1(Beclin1)、LC3、p62、自噬相关蛋白质5(Atg5)、自噬相关16样蛋白1(Atg16L)、溶酶体相关蛋白1(LAMP1)的蛋白质和mRNA的表达。结果与WC组相比,KC组大鼠自梳理次数(P<0.05)及时间(P<0.01)显著增加,埋珠数量显著增加(P<0.01),孔洞探索次数(P<0.05)及单一孔洞探索次数(P<0.01)显著增加,8周游泳运动后,相比于KC组大鼠,KS组大鼠自梳理时间、埋珠数量及单一孔洞探索次数显著降低。此外,与WC组相比,KC组大鼠纹状体区域有大量自噬体形成,同时Atg5(P<0.05)、Atg16L(P<0.01)、p62(P<0.01)的蛋白质及mRNA表达显著上升,LC3II/LC3I比值上升(P<0.01),Beclin1蛋白显著上升(P<0.01),LAMP1的蛋白质及mRNA表达显著下降(P<0.01)。8周游泳运动后,相比于KC组大鼠,KS组大鼠蛋Atg5、Atg16L、p62的蛋白质及mRNA表达显著下降,LC3II/LC3I显著下降(P<0.05),Beclin1蛋白显著下降(P<0.01),LAMP1的蛋白质及m RNA表达显著上升(P<0.05)。结论8周早期游泳可以调节纹状体细胞自噬缓解Shank3^(-/-)大�Objective To explore the mechanism of exercise intervention in improving autism-like behaviors in Shank3 gene knockout(Shank3^(-/-))induced autism spectrum disorder(ASD)rat models from an autophagy perspective through 8-week swimming intervention.Methods Based on genotype identification and exercise intervention,rats were divided into four groups(n=15):wild type control group(WC),Shank3^(-/-)control group(KC),wild-type swimming group(WS),and Shank3^(-/-)swimming group(KS).KS and WS groups underwent 8 weeks of swimming exercise,5 d/week,gradually increasing to and maintaining 40 min/session.Behavioral experiments,including self-grooming test,marble burying test,and hole-board test,were conducted 24 h after the final swimming intervention.Tissue sampling was performed 12 h after behavioral testing.Transmission electron microscopy was used to observe autophagosome numbers in the striatum region.Immunofluorescence staining was employed to observe the expression levels of microtubule-associated protein 1 light chain 3(LC3)and selective autophagy adaptor protein(p62).Quantitative real-time polymerase chain reaction(qPCR)and Western blot were used to detect protein and mRNA expression of Beclin1,LC3,p62,autophagy-related protein 5(Atg5),autophagy-related 16-like protein 1(Atg16L),and lysosome-associated membrane protein 1(LAMP1)in striatal.Results Compared with the WC group,rats in the KC group exhibited significantly higher self-grooming frequency and duration(P<0.05),increased marble burying behavior(P<0.01),and elevated frequencies in both hole-board exploration and single-hole exploration(P<0.05).Following 8 weeks of swimming intervention,the KS group demonstrated significantly reduced self-grooming duration,marble burying behavior,and single-hole exploration frequency compared to the KC group.Furthermore,compared to the WC group,the KC group displayed abundant autophagosomes in the striatum region,along with significantly elevated protein and mRNA expression levels of Atg5,Atg16L,p62,and LC3II/LC3I ratio(P<0.05),in

关 键 词：早期游泳 孤独症谱系障碍 大鼠 细胞自噬 纹状体 

分 类 号：Q95-336[生物学—动物学] R742.89[医药卫生—神经病学与精神病学]