论文摘要
目的:通过腹主动脉插管灌注胶原酶消化分离胰腺星状细胞（pancreatic stellate cell,PSC）,建立简单易行的PSC分离方法,并对PSC进行培养和鉴定;初步探讨PSC早期活化的机制。方法:大鼠腹主动脉插管灌注后,经胶原酶消化、Nycodenz密度梯度离心,分离得到PSC;通过观察细胞形态、胞质内脂滴和免疫细胞化学方法检测结蛋白（desmin）、神经胶质酸纤维蛋白（glial fibrillaryacidic protein,GFAP）、α平滑肌肌动素（α-smooth muscle actin,α-SMA）的表达来鉴定PSC;RT-PCR方法及WestBlot方法测定PSC中Smad2/3表达。结果:腹主动脉灌注法分离得到PSC,产率、活力和纯度分别为:15.3±4.6×103/g体重、95.0±3.5%、＞80%;培养24h后大多数细胞已贴壁,呈星状或多角形,48h后表达desmin、GFAP,并在96h后表达α-SMA;Smad2/3在PSC早期活化中显著高表达,48H后达其高峰。结论:通过腹主动脉灌注法较容易分离得到PSC,其产率、活力和纯度较高,经鉴定后能够满足体外实验要求;Smad2/3的高表达是PSC早期活化过程中的中心环节。Smad2/3的高表达与Smad7无法抑制其活化从而造成两者失去平衡是引起PSC活化的主要原因。
论文目录
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