目的:生物力学力诱导的静脉平滑肌细胞(VSMC)的异常增殖和迁移是限制冠状动脉旁路移植术(CABG)治疗冠心病(CHD)疗效的主要原因。灯盏花素是灯盏花素的主要活性成分,具有广谱的药理作用。因此,本研究旨在研究灯盏乙素在拉伸应力下对VSMCs的影响。
方法:在不同拉伸应力下干扰VSMC后,筛选了最佳拉伸应力。在拉伸应力环境中,100μM黄芩素和橙皮素(p38MAPK途径激活剂)用于用VSMC治疗。CCK-8,EDU,伤口愈合,流式细胞术和蛋白质印迹试验用于检测细胞增殖,迁移,凋亡,以及凋亡相关蛋白(Caspase3、Bcl2和Bax)的表达。
结果:10%的拉伸应力显着增强了活性,伤口愈合率,和VSMC的EDU+细胞,并降低其凋亡率。此外,它上调Bcl2表达,并下调了VSMCs的裂解Caspase3和Bax表达。因此,选择10%的拉伸应力来为VSMC创建拉伸应力环境。有趣的是,100μMScutellarin减轻了10%拉伸应力对VSMC表型的影响。值得注意的是,10%的拉伸应力增加了VSMCs中p38MAPK(Thr180+Tyr182)的磷酸化水平,这是由Scutellarin限制的。Further,橙皮素恢复了灯盏乙素对VSMCs表型的影响。
结论:灯盏乙素通过抑制p38MAPK通路减轻张力应激诱导的VSMCs增殖和迁移。灯盏乙素可作为冠心病患者未来GABG治疗的辅助策略。
OBJECTIVE: Abnormal proliferation and migration of biomechanical force-induced venous smooth muscle cells (VSMCs) is a major cause to limit the efficacy of coronary artery bypass grafting (CABG) for coronary heart disease (CHD). Scutellarin is the main active ingredient of Erigeron Breviscapus, and has broad-spectrum pharmacological effects. Therefore, the present study was proposed to investigate the effect of Scutellarin on VSMCs under tensile stress.
METHODS: After interfering with VSMCs at different tensile stresses, the optimal tensile stress was screened. In a tensile stress environment, 100 μM Scutellarin and Hesperetin (p38 MAPK pathway activator) was used to treatment with VSMCs. CCK-8, EDU, Wound healing, flow cytometry and western blotting assays were used to detect cell proliferation, migration, apoptosis, and the expression of apoptosis-related proteins (Caspase3, Bcl2 and Bax).
RESULTS: Tensile stress with 10% significantly enhanced the activity, wound-healing ratio, and EDU+ cells of VSMCs, and decreased their apoptosis ratio. Moreover, it upregulated Bcl2 expression, and downregulated cleaved-Caspase3 and Bax expression of VSMCs. Hence, 10% tensile stress was selected to creates a tensile stress environment for VSMCs. Interestingly, 100 μM Scutellarin alleviated the effect of 10% tensile stress on the phenotype of VSMCs. Notably, 10% tensile stress increased the phosphorylation level of p38 MAPK (Thr180 +Tyr182) in VSMCs, which was restricted by Scutellarin. Further, Hesperetin restored the effect of Scutellarin on the phenotype of VSMCs.
CONCLUSIONS: Scutellarin alleviates tension stress-induced proliferation and migration of VSMCs via suppressing p38 MAPK pathway. Scutellarin may be used as an adjunctive strategy for future GABG treatment in CHD patients.