Sonic hedgehog促进血管外膜成纤维细胞表型转化并发生增殖迁移

The sonic hedgehog induce vascular adventitial fibroblasts phenotypic modulation , proliferation and migration

目的研究sonic hedgehog（shh）在血管外膜成纤维细胞增殖、迁移、表型转化中的作用。方法体外培养血管外膜成纤维细胞，免疫荧光、激光共聚焦显微成像、免疫印迹以及实时定量聚合酶链式反应检测蛋白和基因表达，Ki67、细胞生长曲线评价细胞增殖情况，划痕实验观察细胞迁移情况，α-肌动蛋白评价细胞表型转化情况。结果shh在体外培养的血管外膜成纤维细胞中有表达，加入外源性的shh后（3．5μg／m1），Ki67^＋的成纤维细胞增多，划痕区域的细胞覆盖率增大、成纤维细胞内出现α-肌动蛋白的表达；加入cyclopamine（40μmol／L）抑制内源性的shh信号通路后，Ki67^＋的成纤维细胞减少，划痕区域的细胞覆盖率减小。结论shh能够促进成纤维细胞增殖、迁移、向肌成纤维细胞转化。

Objective To study the effect of shh on the migration, proliferation and phenotypic modulation of vascular adventitial fibroblasts. Method Cultivate the vascular adventitial fibroblast in vitro. Use immunofluorescent,laser confocal microscopy, Western-blot and real-time PCR to detect the expression of mRNA and protein of related index Estimate cell proliferation according to the expression of Ki67 and cell proliferation curve. Application of wound healing test to estimate migration of fibroblast. The expression of α- actin is thought to be marker of phenotypic modulation of fibroblast. Result The expression of shh was detected in vascular adventitial fibrolast in vitro. After addition of exogenous shh （3.5 μg/ml） , there were： more Ki67 ^＋cells and the wounding area which was covered by ceils became larger. The expression of α-aetin was detected. After addition of cyclopamine（40 μmol/L） , there were less Ki67^ ＋ cells and the wounding area which was covered by cells became smaller. Conclusion Shh can promote proliferation, migration and phenotypic modulation of vascular adventitial fibroblasts.