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目的 本实验旨在探讨多发性硬化(MS)发病中血脑屏障(BBB)中星形胶质细胞(AS)与淋巴细胞(LCs)的相互作用.方法 分离原代鼠脑血管内皮细胞(rBMECs)和SD鼠AS并建立体外BBB模型,对rBMECs与AS共培养和rBMECs常规培养进行跨内皮细胞电阻(TEER)和渗透率的比较.用荧光定量法检测rBMECs单独培养、rBMECs与AS共培养、rBMECs和LCs共培养、rBMECs与AS及LCs三重培养四种模式的神经生长因子(NGF)、基质金属蛋白酶(MMP)-2、MMP-9、白细胞介素(IL)-17,干扰素(IFN)-γ,Foxp3的水平.结果 rBMECs和AS共培养比rBMECs单培养的TEER高,而其LCs渗透率却很低;与rBMECs单培养相比,rBMECs、AS和LCs三重培养可降低NGF、IL-17、IFN-γ的表达(P=0.036),MMP-2和Foxp3的表达增加(P=0.0216);与rBMECs和AS共培养相比,rBMECs、AS、LCs三重培养能有效降低NGF、IL-17和IFN-γ的表达(P =0.0451).与rBMECs和AS的共培养相比,rBMECs、AS、LCs三重培养能增加MMP-2的表达(P =0.021),而MMP-9在四种模式中表达无差异.结论 在体外BBB中原代鼠脑rBMECs和AS共培养比rBMECs单培养的TEER高,AS与LCs共培养可能增加MMP-2的表达,降低IL-17和IFN-γ的表达.“,”Objective The study aimed to investigate the impact of interactions between astrocytes and lymphocytes in blood-brain barrier (BBB) in the pathogenesis of multiple sclerosis (MS).Methods Primary rat brain microvascular endothelial cells (rBMECs) and astrocytes from SD rats were isolated,In vitro BBB model was established.Trans-endothelial electrical resistance (TEER) and permeability were determined.Nerve growth factor (NGF),matrix metalloproteinase (MMP)-2,MMP-9,interleukin (IL)-17,interferon (IFN)-γ,and forkhead box P3 (Foxp3) were measured by fluorescent real-time quantitative polymerase chain reaction (PCR) in 4 models:rBMECs mono-culture,rBMECs and astrocyte co-Culture,rBMECs and lymphocyte co-Culture,and rBMECs,astrocytes and lymphocyte tri-culture.Results rBMECs and astrocytes co-culture had higher TEER values and lower permeability of lymphocytes than rBMEC mono-culture.Compared with rBMEC mono-culture,rBMEC,astrocytes and lymphocytes tri-cultire had significantly decreased level of NGF,IL-17,IFN-γ(P =0.036),MMP-2,and Foxp3 (P =0.0216).In comparison with rBMEC and astrocyte co-culture,the tri-culture had further decreased level of NGF,IL-17,and IFN-γ(P =0.0451).The expression of MMP-2 was further increased in the tri-culture model compared to that of rBMEC and lymphocyte co-culture(P =0.021).The expression of MMP-9 was not significantly differentamong the 4 models.Conclusion The study successfully established in vitro BBB model with primary rBMECs and astrocyte co-culture.The research speculated that interaction between astrocytes and lymphocytes might increase the expression of MMP-2 and decrease the expression of IL-17 and IFN-γ.