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目的在体外模拟血管外壁组织结构,建立一种可应用于研究氧合血红蛋白(Oxy- Hb)诱导的脑血管痉挛的新型细胞模型。方法以微孔多聚碳酸酯膜(PET膜)作为载体,模拟血管壁外弹力层,将成纤维细胞(AFB)和平滑肌细胞(SMC)分别接种于PET膜的两侧共培养,建立AFB-PET-SMC血管外壁重构模型。在PET膜一侧接种的AFB中加入含1×10~(-6)mol/L OxyHb的培养液,而另一侧的SMC培养液中无OxyHb,分别共培养24、48、72 h,应用扫描电镜观察SMC的长度。结果AFB-PET-SMC组织结构关系类似于体内血管外层(成纤维细胞-外弹力层-平滑肌细胞)结构；模拟体内出血环境,OxyHb处理AFB 24、48、72 h后SMC平均长度分别为(41．6±9．21)、(28．34±8．38)、(19．80±7．09)μm,与正常组(55．66±10．35)μm比较差异有统计学意义(P＜0．01)。结论OxyHb作用于PET膜一侧的AFB引起对侧未直接接触OxyHb的SMC发生持续收缩,该细胞模型可应用于研究OxyHb诱导的脑血管痉挛。
Objective To simulate the structure of the vascular wall in vitro and establish a new cell model that can be used to study the cerebral vasospasm induced by oxyhemoglobin (Oxy-Hb). Methods Microporous polycarbonate (PET) film was used as a carrier to simulate the elastic layer of vascular wall. AFB and SMC were inoculated on both sides of PET film to establish AFB-PET -SMC vascular remodeling model. The AFB containing 1 × 10 -6 mol / L OxyHb was inoculated on the side of PET film, while the OxyHb was not cultured in the other SMC medium for 24, 48 and 72 h respectively. Scanning electron microscopy SMC length. RESULTS: The tissue structure of AFB-PET-SMC was similar to that of the vascular endothelial layer (fibroblast-EMR-smooth muscle cell) in vivo. Simulating the hemorrhagic environment in vivo, the average length of SMC after OxyHb treatment for AFB for 24, 48 and 72 h were 41.6 ± 9.21), (28.34 ± 8.38) and (19.80 ± 7.09) μm, respectively, which were significantly different from those of the normal group (55.66 ± 10.35) μm P <0.01). CONCLUSIONS: OxyHb acts on the PET film side of the AFB, causing contralateral non-direct contact OxyHb SMC continued contraction, the cell model can be used to study OxyHb-induced cerebral vasospasm.