Kudo Fabio A, Warycha Bohdan, Juran Peter J, Asada Hidenori, Teso Desarom, Aziz Faisal, Frattini Jared, Sumpio Bauer E, Nishibe Toshiya, Cha Charles, Dardik Alan
Department of Surgery, Yale University School of Medicine, Boyer Center for Molecular Medicine, 295 Congress Ave., New Haven, CT 06519, USA.
Am J Surg. 2005 Nov;190(5):763-9. doi: 10.1016/j.amjsurg.2005.07.017.
The incidence of vascular disease increases with age. Because atherosclerosis and neointimal hyperplasia colocalize in areas of disturbed shear stress, the effects of orbital shear stress (SS) on endothelial cell proliferation, protein kinase B (Akt) activation, and functional activity were analyzed using a senescence model.
Early- (p3 to 7) and late- (p28 to 32) passage bovine aortic endothelial cells were exposed to orbital SS (210 rpm) or static conditions (0 to 5 days). Cell proliferation was directly counted and confirmed with proliferating cell nuclear antigen reactivity. Phosphorylated and total Akt were assessed with Western blotting. Endothelial cell-induced smooth muscle cell migration was assessed with a Boyden chamber.
Late-passage endothelial cells demonstrated no increase in orbital SS stimulated proliferation compared with early-passage cells (P = .42). Late-passage endothelial cells demonstrated decreased Akt phosphorylation in response to SS compared with early passage cells (n = 6, P = .01). Late-passage cells induced 26% less smooth muscle cell migration than early-passage cells (n = 3, P = .03).
Late-passage endothelial cells demonstrate decreased proliferation, Akt phosphorylation, and secretion of smooth muscle cell chemoattractants in response to orbital SS compared with early passage cells. These results suggest that late-passage endothelial cells respond to SS differently than early-passage cells and confirm the utility of the in vitro senescence model.
血管疾病的发病率随年龄增长而增加。由于动脉粥样硬化和新生内膜增生共同存在于剪切应力紊乱的区域，因此使用衰老模型分析了眼眶剪切应力（SS）对内皮细胞增殖、蛋白激酶B（Akt）激活和功能活性的影响。
将传代早期（第3至7代）和传代后期（第28至32代）的牛主动脉内皮细胞暴露于眼眶SS（210转/分钟）或静态条件下（0至5天）。直接计数细胞增殖情况，并用增殖细胞核抗原反应性进行确认。通过蛋白质印迹法评估磷酸化Akt和总Akt。使用博伊登小室评估内皮细胞诱导的平滑肌细胞迁移。
与传代早期细胞相比，传代后期内皮细胞在眼眶SS刺激下的增殖没有增加（P = 0.42）。与传代早期细胞相比，传代后期内皮细胞对SS的反应中Akt磷酸化降低（n = 6，P = 0.01）。传代后期细胞诱导的平滑肌细胞迁移比传代早期细胞少26%（n = 3，P = 0.03）。
与传代早期细胞相比，传代后期内皮细胞在眼眶SS刺激下表现出增殖减少、Akt磷酸化降低以及平滑肌细胞趋化因子分泌减少。这些结果表明，传代后期内皮细胞对SS的反应与传代早期细胞不同，并证实了体外衰老模型的实用性。
