自噬在实验性自身免疫性脑脊髓炎小鼠中的抗炎作用 - 第三军医大学学报.doc

自噬在实验性自身免疫性脑脊髓炎小鼠中的抗炎作用 许梦雪，秦新月，冯金洲，燕伟平，张融融（400016重庆，重庆医科大学附属第一医院神经内科，重庆市神经病学重点实验室）（antophagy）EAE病程中的时序变化，并初步探讨其对小鼠实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitisEAE）C57BL/6雌性小鼠建模后分别于3、7、16、30 Western blot检测CNS中LC3B的变化。另将60只小鼠随机分为正常对照组、EAE模型组、EAE+3-Methyladenine（3-MA）组、EAE+PBS组。神经功能评分观察各组小鼠神经功能变化。免疫组化检测各组小鼠免疫后7 d小胶质细胞激活情况，Western blot检测免疫后16、30 dβ（IL-1β）的表达，HE染色观察CNS炎性细胞浸润情况。结果 ①EAE+3-MA组小鼠神经功能缺损症状较其他组严重（P0.05）-II/LC3-I自免疫后7 d开始降低（P0.05），此后一直呈低表达水平（P0.0）③EAE免疫后7 d即有小胶质细胞激活，EAE+3-MA组EAE小鼠小胶质细胞激活程度较EAE模型组严重（P0.05）HE染色结果显EAE+3-MA组炎细胞浸润比同时间点EAE模型组加重（P0.05）16、30 dEAE模型组高表达IL-1β，EAE+3-MA组小鼠IL-1β表达水平较同时间点EAE模型组高（P0.05）。结论 EAE中，早期即存在着自噬水平降低。进一步抑制自噬会促进疾病早期小胶质细胞激活，升高病程中IL-1β的表达，加重神经功能缺损，故EAE自噬水平的降低是导致疾病炎症反应的可能原因。 [关键词]IL-1β [基金项目年份？）nti-inflamatory effect of autophagy in experimental autoimmune encephalomyelitis mice Xu Mengxue, Qin Xinyue, Feng Jinzhou, Yan Weiping, Zhang Rongrong (Department of Neurology,First Affiliated Hospital,Chongqing Medical University,Chongqing, 400016, China) [Abstract] Objective To explore autophagy in EAE C57BL/6 mice at different time points post-immunization, and demonstrate its anti-inflamatory effect in EAE mice. Methods EAE models were established in 20 female C57BL/6 mice and then were killed on 3、7、16、30 days later for the expression of LC3B in the CNS by Western blot. Another 60 female C57BL/6 mice were randomly divided into the normal control group, EAE group, EAE+3-Methyladenine（3-MA）group and EAE+PBS group. Neurological function scores were performed to evaluate the neurological function. Immunohistochemistry was performed to test the activation of microgliapost immunization. Western blot was performed to observe the expression of IL-1β in each group on 16d and 30d post immunization. H-E stain was used to observe the extend of inflammatory infiltration in the brain. Results ①Western blot showed the expression of LC3 -II/LC-3I decreased at 7 days post immunization((P0.05)，and keep in a lower level compared to the normal gro