中枢神经系统周细胞表达结蛋白desmin

目的:本研究着重探讨中枢神经系统周细胞是否表达desmin。方法:取3周龄雄性Wistar大鼠,取其大脑和脊髓组织,分离提取培养脑微血管周细胞(BMP)和脊髓微血管周细胞(SCMP)两种周细胞,用周细胞非特异性标记物神经胶质抗原2(NG2)和α-平滑肌肌动蛋白(α-SMA)双标鉴定周细胞,并用免疫荧光方法定性定量测定两种周细胞表达desmin阳性率,免疫印记分析实验定量测定两种周细胞表达desmin。结果:观察刚分离得到的脑微血管和脊髓微血管,前者在长度和密度上多于后者,培养至第3天时,观察到BMP更趋向于聚集,而SCMP则更分散。培养至第9天时,两种周细胞基本铺满了培养皿。经用周细胞非特异性标记物NG2和α-SMA双标鉴定,确定分离得到的细胞为周细胞,免疫荧光和western blot实验结果表明两种周细胞均表达desmin,且SCMP表达desmin阳性率显著多于BMP(P0.001),SCMP表达desmin量显著多于BMP(P0.05)。结论:中枢神经系统周细胞表达结蛋白desmin,表达量不同暗示了血脑屏障和血脊髓屏障之间存在差异。

Pericytes fromCentral Nervous SystemExpressed Desmin

Objective:This research is to observe whether CNS pericytes express desmin.Methods:Pericytes were isolated frombrain and spinal cord, which were taken from 3-week-old Wistar male rat. The cultured pericytes were identified by nonspecific markerNG2 and alpha-SMA. Then the positive rate of pericytes which expressed desmin was determined by immunofluorescence assay and thedesmin expression quantity of pericytes was determined by western blot assay.Results:It was observed that the length and density ofisolated brain microvascular was more than the spinal cord microvascular. On the third day, it was observed that BMP gathered together,while SCMP scattered. On the ninth day, the dishes were almost carpeted by BMP and SCMP. It was confirmed that the isolated cellswere pericytes by the nonspecific biomarkers NG2 and alpha-SMA. Both of the pericytes expressed desmin. The positive rate of SCMPexpressed desmin more than that of BMP (P<0.001). The desmin expression quantity of SCMP was more than BMP (P<0.05).Conclusion:CNS pericytes expressed different amount of desmin, which indicated the differences between blood brain barrier and bloodspinal cord barrier.