免疫启动前烟草烟雾暴露促进EAE发病的细胞免疫机制初步研究  

Effect of tobacco smoke exposure before immune initiation on pathogenesis of experimental autoimmune encephalomyelitis in rats

作　　者：柴嘉擎 王赵伟[2] 吴承龙[2] 蒋琦 王鑫[4] CHAI Jiaqing;WANG Zhaowei;WU Chenglong;JIANG Qi;WANG Xin(School of Medicine,Shaoxing University,Shaoxing 312000,China;不详)

机构地区：[1]绍兴文理学院医学院,312000 [2]绍兴市人民医院神经内科 [3]绍兴市人民医院输血科 [4]绍兴市人民医院风湿免疫科

出　　处：《浙江医学》2023年第7期680-684,I0003,共6页Zhejiang Medical Journal

基　　金：浙江省基础公益研究计划项目(LGD22H090016、LGF18H090018);绍兴市卫生健康科技计划项目(2022KY015);绍兴市人民医院2022年度青年科研基金项目(2022YB26)。

摘　　要：目的探讨自身免疫激活前短暂烟草烟雾暴露对成年SD大鼠实验性自身免疫性脑脊髓炎(EAE)发病及脊髓炎症反应的影响。方法采用随机数字表法将30只SD大鼠分为空白对照组、空气组和烟草组。在EAE造模前1周予以连续烟草烟雾暴露7 d,观察各组动物神经行为学变化并评分。在EAE造模后第21天抽取脑脊液并处死动物,采用ELISA法检测脑脊液IL-17F、IL-10水平,流式细胞术检测脾脏CD4^(+)T细胞亚型变化。颈髓、胸髓、腰髓石蜡切片经HE染色、监牢蓝髓鞘染色后评估炎症反应及脱髓鞘百分比。结果烟草组最高行为学评分高于空气组[3.50(1.00,4.00)分比2.00(0,2.25)分],差异有统计学意义(P<0.05)。烟草组颈髓、胸髓、腰髓HE染色病理评分和脱髓鞘百分比均高于空气组,差异均有统计学意义(均P<0.05)。烟草组抗炎的CD4^(+)叉头蛋白P3(FOXP3)^(+)T细胞百分比低于空气组,而具有致病性的CD4^(+)FOXP3^(-)T细胞百分比高于空气组,差异均有统计学意义(均P<0.05)。烟草组脑脊液IL-17F水平高于空气组,IL-10水平低于空气组,差异均有统计学意义(均P<0.05)。结论免疫激活前烟草烟雾暴露可加重成年大鼠EAE神经功能受损,并促进脊髓炎症反应。Objective To investigate the effect of short-term tobacco smoke exposure before autoimmune activation on the onset of experimental autoimmune encephalomyelitis(EAE)in rats.Methods Thirty SD rats were randomly divided into blank control group,air group and tobacco group.The animals were exposed or not exposed to tobacco smoke for 7 days 1 week before EAE modeling in tobacco group and air group,respectively.The neurobehavioral changes were observed and scored in each group.The animals were sacrificed on day 21 after EAE modeling,the cerebrospinal fluid was extracted and the concentrations of IL-17F and IL-10 in the cerebrospinal fluid were measured by ELISA,the splenic CD4^(+)T cell subtypes were detected by flow cytometry.Paraffin sections of the cervical,thoracic and lumbar medulla were stained with HE and luxol fast blue(LFB)myelin staining to assess the inflammatory response and the percentage of demyelination.Results The top behavioral score was higher in the tobacco group than that in the air group[3.50(1.00,4.00)vs.2.00(0,2.25)points,P<0.05].The cervical,thoracic,and lumbar medulla HE staining scores and the percentage of demyelination were higher in the tobacco group than those in the air group(all P<0.05).The percentage of CD4^(+)FOXP3^(+)T cells was lower in the tobacco group than that in the air group,while the percentage of CD4^(+)FOXP3^(-)T cells was higher than that in the air group(all P<0.05).Cerebrospinal fluid IL-17F levels were higher in the tobacco group than that in the air group,and IL-10 levels were lower than that in the air group(all P<0.05).Conclusion Pre-immune activation tobacco smoke exposure exacerbates neurological impairment and promotes spinal cord inflammatory responses in adult rats with EAE.

关 键 词：烟草烟雾 实验性自身免疫性脑脊髓炎 多发性硬化 T细胞 

分 类 号：R744.51[医药卫生—神经病学与精神病学]