小鼠青春期香烟暴露通过前额叶小胶质细胞介导的炎症反应影响成年后学习记忆功能  

Smoke exposure during adolescence affects learning and memory abilities of mice in adulthood via prefrontal microglia-mediated inflammation

作　　者：范文娟[1,2] 陈彦锋 段子朋[1] 孙缦利[1] 陈旭东[1] FAN Wenjuan;CHEN Yanfeng;DUAN Zipeng;SUN Manli;CHEN Xudong(Luohe Medical College,Henan Provincial Engineering Research Center of Nutrition and Health,Luohe 462002,China;Department of Neurology,Luohe Central People's Hospital,Luohe 462002,China)

机构地区：[1]漯河医学高等专科学校,河南省营养与健康工程研究中心,河南漯河462002 [2]漯河市中心医院神经内科,河南漯河462002

出　　处：《中国病理生理杂志》2024年第12期2254-2261,共8页Chinese Journal of Pathophysiology

基　　金：河南省自然科学基金资助项目(No.222300420246);河南省重点研发与推广专项(No.232102310501);漯河医专科技创新项目(No.2023ZD08,No.2023ZD04)。

摘　　要：目的:探讨小鼠青春期香烟暴露对前额叶小胶质细胞介导的炎症反应及其对成年后学习记忆功能的影响。方法:健康2周龄雄性昆明小鼠72只,每只体重(11.0±1.5) g。随机分为对照组与香烟暴露组,每组各36只。香烟暴露组每日被动吸烟6支,持续10周。分别在对照组与香烟暴露组小鼠4周龄(少年期)、8周龄(青年期)、12周龄(成年期)三个时点,每组各取6只检测其神经行为学与病理学变化。具体如下:通过跳台测试、三箱交互实验、新奇事物识别实验检测小鼠学习记忆能力及认知行为学功能变化;免疫荧光实验检测各组小鼠前额叶皮质小胶质细胞的形态、突触数量以及小胶质细胞周围炎性因子凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD, ASC)的表达;Western blot实验检测各组小鼠大脑皮质突触小泡蛋白(synaptophysin,SYP)和突触后致密蛋白95(postsynaptic density protein-95, PSD-95)的表达水平;酶联免疫吸附测定(enzyme-linked immunosorbent assay, ELISA)实验检测前额叶皮质中肿瘤坏死因子α (tumor necrosis factor-α, TNF-α)、白细胞介素1β(interleukin-1β, IL-1β)、IL-6的蛋白表达。结果:(1)与同龄对照组相比,8周、12周龄香烟暴露小鼠跳台潜伏期显著缩短、跳台错误次数显著增多(P<0.01),社交识别时间以及对新事物探索时间延长(P<0.05)。(2)免疫荧光实验表明,8周龄、12周龄香烟暴露组小鼠前额叶皮质SYP阳性斑点减少,Iba1阳性小胶质细胞数量增多,呈现激活状态;小胶质细胞周围ASC阳性斑点增多。Western blot实验进一步证明8周龄、12周龄香烟暴露组小鼠大脑皮质突触素蛋白SYP、PSD-95表达降低(P<0.05)。(3) ELISA实验表明8周龄、12周龄香烟暴露组小鼠前额叶皮质炎性因子TNF-α、IL-1β和IL-6含量增高(P<0.05)。结论:小鼠青春期香烟暴露可能会通过激活前额叶皮质小胶质细胞,导致炎症因子分泌增多,影AIM:To investigate the effects of cigarette smoke exposure during adolescence on the inflammatory response mediated by microglia in the prefrontal cortex of mice,and its impact on learning and memory functions in adulthood.METHODS:72 two-week-old healthy male Kunming mice,with each weighing(11.0±1.5)g,were randomly divided into control and cigarette exposure groups(n=36 per group).The mice in the cigarette exposure group were passively exposed to 6 cigarettes daily for 10 weeks.At three time points of 4-week-old(infancy),8-week-old(adolescence),and 12-week-old(adulthood),six mice were selected from each group to have their neurobehavioral and pathological changes examined.In particular,the step-down test,three-chamber social interaction test,and novel object recognition test were used to detect changes in learning and memory abilities and cognitive behavior.Immunofluorescence testing was performed to detect the morphology, number of synapses, and expression of inflammatory factor apoptosis-associated speck-like protein containing a CARD (ASC) around the microglial cells in the prefrontal cortex of mice in each group. Western blot was performed to assess the expression levels of synaptophysin (SYP) and postsynaptic density protein-95 (PSD-95) in the cerebral cortex of mice in each group. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the protein expression of tumor necrosis factor-α (TNF-α), interleukin 1β( IL-1β), and IL-6 in the prefrontal cor-tex. RESULTS:( 1) In the step-down test, the latency of mice at 8 and 12 weeks of age was significantly shortened, and the number of errors was significantly increased in the cigarette exposure group compared with the age-matched control group (P<0. 01). In addition, the social recognition time and exploration time for novel objects were prolonged (P< 0. 05).( 2) Immunofluorescence assays revealed that exposure to cigarette smoke in mice, at both 8 and 12 weeks of age, resulted in a reduction of SYP-positive puncta within the prefrontal cortex. Conc

关 键 词：香烟暴露 前额叶皮质 小胶质细胞 认知 炎症 

分 类 号：R742[医药卫生—神经病学与精神病学] R338.6[医药卫生—临床医学] Q421[生物学—神经生物学]