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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(P<0.001),SCMP表达desmin量显著多于BMP(P<0.05)。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。
Aims: This study focuses on whether central nervous system pericytes express desmin. Methods: Male Wistar rats, aged 3 weeks, were taken from the brain and spinal cord. Two types of pericytes, cerebral capillary pericytes (SCMP) and cerebral microvessel pericytes (BMPs), were isolated and extracted. Pericytes Pericytes were identified by double labeling of NG2 and α-SMA, and the expression of desmin was qualitatively and quantitatively detected by immunofluorescence. The expression of two kinds of pericytes was quantified by Western blot analysis Expression desmin. RESULTS: The isolated brain microvessels and splenic microvessels were observed. The former was more in length and density than the latter. When cultured to day 3, it was observed that BMP tended to aggregate and SCMP more dispersed. On day 9 of culturing, both pericytes were predominantly covered with petri dishes. The results of immunofluorescence and western blot showed that both of the two kinds of pericytes expressed desmin, and the positive rate of desmin in SCMP was significantly higher In BMP (P <0.001), the expression of SCMP in desmin was significantly higher than that in BMP (P <0.05). CONCLUSIONS: The expression of desmin in the central nervous system pericytes expressed differently, suggesting a difference between the blood-brain barrier and the blood-spinal cord barrier.