一种可应用于研究脑血管痉挛的新型细胞模型

A novel in vitro model applied in the study of oxyhemoglobin-induced cerebral vasospasm

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 establish a novel in vitro model imitating part of the vascular wall that can be appiied in the study of cerebral vasospasm induced by oxyhemoglobin （ OxyHb ）： Methods Smooth muscle cells （SMC） and adventitial fibroblasts （AFB） grew each on opposite sides ot microporous polycarbonate filters （PET）, to establish a SMC-PET-AFB cell model imitating the outer layer ot vascular wall. The AFB cells were incubated in the medium containing 10^-6 mol/L oxyhemoglobin, while thesmooth muscle cells in the opposite side incubated without oxyhemoglobin. After the two types of the cells were cocultured for 24,48 and 72 h,the length changes of SMC were examined under an electron micro- scope. Results A SMC-PET-AFB model was successfully established, which was similar to the outer layer of the vascular wall structure in vivo. The mean length of SMC in 24,48 and 72 h groups was （41.6 ± 9.21 ）, （28.34 ± 8.38） and （ 19.80 ± 7.09） μm respectively, significantly shorter than that of normal group [（55.66± 10.35） μm （P〈0.01）].Conelusion The AFB cells incubated with oxyhemoglobin can induce the contraction of the cocultured SMC incubated without oxyhemoglobin. The novel in vitro model can be applied in the study of oxyhemoglobin-induced cerebral vasospasm.