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目的探讨切应力与血管平滑肌细胞(VSMCs)对内皮细胞(ECs)增殖功能的影响及其一些分子机制。方法应用平行平板流动腔系统对单独培养的ECs以及与VSMCs联合培养的ECs施加15dyn/cm2(1dyn=10-5N)的正常切应力;Western blot技术检测反映细胞增殖能力的分子—增殖细胞核抗原(PCNA)的表达及细胞内信号转导分子Akt的磷酸化水平。静态条件下,以单独培养ECs为对照组,将ECs与VSMCs隔开培养,并应用TGFβ1封闭性抗体,观察TGFβ1在VSMCs诱导ECs增殖中的作用。结果正常切应力抑制了ECs增殖及p-Akt表达,VSMCs在与ECs联合培养及隔开培养时均明显促进ECs增殖及p-Akt表达。正常切应力部分逆转了联合培养VSMCs诱导的ECs增殖和p-Akt表达,而TGFβ1封闭性抗体能够拮抗隔开培养VSMCs诱导的ECs增殖和p-Akt表达。结论正常切应力可视为血管的保护因子,抑制ECs增殖;VSMCs通过旁分泌作用诱导了ECs增殖;TGFβ1及PI3/Akt信号分子参与了其调节过程。
Objective To investigate the effects of shear stress and vascular smooth muscle cells (VSMCs) on the proliferation of endothelial cells (ECs) and some of its molecular mechanisms. Methods Normal shear stress of 15dyn / cm2 (1dyn = 10-5N) was applied to cultured ECs and ECs co-cultured with VSMCs using parallel plate flow chamber system. Western blot was used to detect the expression of molecular proliferating cell nuclear antigen ( PCNA) expression and phosphorylation of intracellular signal transduction molecule Akt. Under static conditions, ECs were cultured as a control group, ECs were cultured separately from VSMCs, and TGFβ1 blocking antibodies were applied to observe the effect of TGFβ1 on the proliferation of ECs induced by VSMCs. Results Normal shear stress inhibited the proliferation of ECs and the expression of p-Akt. VSMCs significantly promoted the proliferation of ECs and the expression of p-Akt when cultured with ECs and cultured separately. Normal shear stress partially reversed the ECs proliferation and p-Akt expression induced by co-cultured VSMCs, whereas TGFβ1 blocking antibodies antagonized the proliferation of ECs induced by VSMCs and the expression of p-Akt. Conclusion Normal shear stress can be regarded as a protective factor of blood vessel and inhibit the proliferation of ECs. VSMCs induce ECs proliferation through paracrine effect. TGFβ1 and PI3 / Akt signaling molecules participate in the regulation.