星形胶质细胞及淋巴细胞在多发性硬化血脑屏障中作用机制研究  

Mechanism of astrocytes and lymphocytes in the blood-brain barrier of multiple sclerosis

作　　者：闫晓波[1] 李晨晔[1] 邱晓恬 赵延峰[1] 杨永梅[1] Yah Xiaobo;Li ChenYe;Qiu Xiaotian;Zhao Yanfeng;Yan yongmei(Department of Neurology,the Second Affiliated Hospital of Harbin Medical University,Harbin 150086,China)

机构地区：[1]哈尔滨医科大学附属第二医院神经内科,150086

出　　处：《国际免疫学杂志》2018年第6期620-624,共5页International Journal of Immunology

摘　　要：目的 本研究旨在探索星形胶质细胞(astrocyte,AS)和淋巴细胞(lymphocytes,LCs)在多发性硬化(multiple sclerosis,MS)血脑屏障(blood brain barrier,BBB)受损中的作用.方法 分离初代大鼠脑微血管内皮细胞(rat brain microvascular endothelial cell,rBMECs)和鼠AS并建立体外BBB模型.通过比较rBMECs单培养和其与AS混合培养组的跨内皮细胞电阻(transendothelial cell resistance,TEER)和渗透性来评估正常BBB的细胞模式.在rBMECs和LCs共培养组和rBMECs、AS、LCs共培养组中通过对神经生长因子(nerve growth factor,NGF)、基质金属蛋白酶(matrix metalloproteinase,MMP)-2、MMP-9、白细胞介素(interleukin,IL)-17、干扰素(interferon-γ,IFN-γ)、FoxP3的mRNA表达水平的检测进一步研究MS的炎症反应特点.结果rBMECs和AS共培养组比rBMECs单培养组的TEER高(>300Ωcm2),而其LCs渗透率却更低(<200000).与rBMECs单培养组相比,rBMECs、AS、LCs共培养组的NGF、IL-17、IFN-γ的表达降低,而MMP-2和Foxp3表达增高,差异均有统计学意义(P=0.036,0.022).与rBMECs和AS的共培养相比,rBMECs、AS、LCs共培养组MMP-2的表达增加,NGF,IL-17和IFN-γ的表达降低.结论AS和LCs之间相互作用的可能会增加MMP-2的表达,降低IL-17、IFN-γ的表达.提示AS与LCs的协同作用可能参与了MS的发病.Objective To investigating the interactions between astrocytes (AS)and lymphocytes (LCs)at the blood-brain barrier(BBB)on the pathogenesis of multiple sclerosis (MS). Methods Primary rat brain microvascular endothelial cells (rBMECs)and AS isolated from SD rats were used to establish BBB mod-els in vitro. Trans-endothelial electrical resistance (TEER)and permeability were compared between an rBMEC mono-culture and rBMEC/ AS co-culture,to evaluate the validity of each as a BBB cell model. rBMEC/ LCs co-culture and rBMEC/ AS/ LCs tri-culture were established to evaluate the inflammatory response in MS by meas-uring the expression of nerve growth factor (NGF),matrix metalloproteinase (MMP)-2,MMP-9,interleukin (IL)-17,interferon γ (IFN-γ),and forkhead box P3 (Foxp3). Results The rBMEC/ AS co-culture was ex-hibited a higher TEER value(> 300 Ωcm2 )and lower lymphocyte permeability(< 200000)than did the rBMEC mono-culture. Compared to the rBMEC mono-culture,the rBMEC/ AS/ LCs tri-culture showed signifi-cantly decreased the expression of NGF,IL-17,and IFN-γ and increased the expression of MMP-2 and Foxp3 (P =0. 036;P =0. 022). Conclusion The results suggest that the interaction between AS and LCs may increase the expression of MMP-2 and decrease expression of IL-17 and IFN-γ at the BBB. Furthermore,the rBMEC/ AS/LCs tri-culture promted the synergistic effect between AS and LCS may be involved in the pathogenesis of MS.

关 键 词：血脑屏障 脑微血管内皮细胞 跨内皮细胞电阻 

分 类 号：R744.51[医药卫生—神经病学与精神病学]