纳米炭黑暴露对小鼠神经行为损伤及Th17细胞浸润的影响  

Neurobehavioral damage and Th17 cell infiltration in mice exposed to nano carbon black

作　　者：郝瀚 朱皓 张荣[2] 王伟轩 张艳淑 Hao Han;Zhu Hao;Zhang Rong;Wang Weixuan;Zhang Yanshu(School of Public Health,North China University of Science and Technology,Tangshan 063210,China;School of Public Health,Hebei Medical University,Shijiazhuang050017,China;Laboratory Animal Center,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]华北理工大学公共卫生学院,唐山063210 [2]河北医科大学公共卫生学院,石家庄050017 [3]华北理工大学实验动物中心,唐山063210

出　　处：《卫生研究》2023年第5期702-709,共8页Journal of Hygiene Research

基　　金：国家自然科学基金(No.82073598);河北省教育厅科技项目(No.ZD2020113)。

摘　　要：目的通过建立亚急性动式吸入炭黑气溶胶小鼠染毒模型,探讨纳米炭黑对小鼠神经行为的损伤作用以及Th17细胞浸润的影响。方法SPF级雄性C57BL/6小鼠36只,随机分为对照组(过滤洁净空气)、低炭黑组(15 mg/m^(3))和高炭黑组(30 mg/m^(3))。利用气溶胶发生器将纳米炭黑颗粒吹入动式染毒柜中,连续染毒28 d。应用Morris水迷宫实验和旷场实验检测小鼠神经行为变化,采用HE染色法观察小鼠前额叶皮质的病理变化,应用流式细胞术检测小鼠外周血及脑组织中Th17/CD4^(+)细胞比例的变化,应用Western blotting检测小鼠前额叶皮质中白细胞介素(interleukin,IL)-17和IL-23蛋白表达的变化。结果旷场实验结果显示,与对照组比较,低、高炭黑组小鼠中央区域停留时间和站立次数显著下降(P<0.05),其中高炭黑组排便次数显著增高(P<0.05),高炭黑组中央区域停留时间显著低于低炭黑组(P<0.05)。Morris水迷宫结果显示,高炭黑组小鼠第3天的逃逸潜伏期显著高于对照组(P<0.05),各炭黑组小鼠第4天的逃逸潜伏期均显著高于对照组(P<0.05)。定位航行实验发现,高炭黑组小鼠穿台次数显著高于对照组(P<0.05)。HE染色结果显示,对照组小鼠前额叶皮质中神经细胞圆润,细胞质饱满分布均匀且细胞核呈椭圆形清晰可见;低炭黑组出现神经细胞深染,结构模糊,核固缩现象;高炭黑组则进一步加剧。流式细胞术结果显示,与对照组比较,各炭黑组小鼠外周血中Th17/CD4^(+)T细胞的百分比显著升高(P<0.05),高炭黑组小鼠显著高于低炭黑组(P<0.05);各炭黑组小鼠脑组织中Th17/CD4^(+)T细胞的百分比显著升高(P<0.05);高炭黑组显著高于低炭黑组(P<0.05)。Western blotting结果显示,与对照组比较,各炭黑组小鼠脑组织前额叶皮质中IL-17、IL-23蛋白表达显著升高(P<0.05);与低炭黑组比较,高炭黑组小鼠脑组织前额叶皮质中IL-17、IL-23蛋白表达显著升高(P<0.05)�OBJECTIVE The effects of nano-carbon black on neural behavior and Th17 cell infiltration in mice were investigated by establishing a mice model of subacute dynamic inhalation of carbon black aerosol.METHODS 36 SPF grade male C57BL/6 mice were randomly divided into a control group(clean air),a low carbon black group(15 mg/m^(3)),and a high carbon black group(30 mg/m^(3)).Nano-carbon black particles were blown into the dynamic exposure cabinet by aerosol generator for 28 days.Morris water maze test and open field test were used to detect the neural behavior of mice.The pathological changes of prefrontal cortex in mice were observed by HE staining.The proportion of Th17/CD4^(+)cells in peripheral blood and brain tissue of mice was detected by flow cytometry.Western blotting was used to detect the protein expression of interleukin(IL)-17 and IL-23 in the prefrontal cortex of mice.RESULTS The result of open field test showed that compared with the control group,the central area residence time and standing times of mice in the low and high carbon black groups decreased significantly(P<0.05),and the defecation times of mice in the high carbon black group increased significantly(P<0.05).The central area residence time of mice in the high carbon black group was significantly lower than that in the low carbon black group(P<0.05).The Morris water maze result showed that the escape latency of the high carbon black group mice on the 3rd day was significantly higher than that of the control group(P<0.05).Meanwhile,the escape latency of the carbon black group mice on the 4th day was significantly higher than that of the control group(P<0.05).The positioning navigation test showed that the number of mice crossing the platform in the high carbon black group was significantly higher than that in the control group(P<0.05).The HE staining result showed that the neural cells in the prefrontal cortex of the control group mice were round,the cytoplasm was plump and evenly distributed,and the nucleus was clearly visible in an oval shape

关 键 词：炭黑颗粒 TH17 白细胞介素-17 白细胞介素-23 神经炎症 

分 类 号：R994.6[医药卫生—毒理学] R135[医药卫生—药学]