摘要
目的 :观察7-二氟亚甲基-5,4-二甲氧基异黄酮(DFMG)是否能干预损伤内皮细胞对平滑肌细胞增殖和迁移的促进作用,且其干预过程是否与TLR4信号通路相关。方法 :采用CCK-8法和Transwell迁移法测定LPC诱导的内皮细胞的损伤对平滑肌细胞的增殖和迁移的影响。采用Western blot和荧光定量PCR测定损伤共培养体系中的内皮细胞TLR4在蛋白水平和基因水平的表达。运用TLR4特异性激动剂(LPS)和特异性的抑制剂(CLI095),采用CCK8法和Transwell迁移法测定损伤共培养体系中平滑肌细胞的增殖能力和迁移能力。结果 :(1)随着LPC浓度的增加,LPC诱导的内皮细胞损伤引起平滑肌细胞的增殖和迁移的效应增强,并且选取30μM的LPC作用于内皮细胞并与平滑肌细胞共培养作为实验的损伤共培养模型。(2)经光密度值分析,LPC组的内皮细胞TLR4的表达量是对照组的2倍;经荧光定量PCR分析,LPC组的内皮细胞TLR4的表达量是对照组的2.414倍。(3)相比,损伤组和LPS组的平滑肌细胞的增殖和迁移能力较对照组更强;相比,CLI095组和DFMG组的平滑肌细胞的增殖和迁移能力较损伤组更弱。结论 :LPC诱导的内皮细胞的损伤可以引起平滑肌细胞的增殖和迁移,而DFMG可能是通过抑制TLR4信号阻碍损伤的内皮细胞对平滑肌细胞增殖和迁移的促进作用。
Objective To observe the effect of 7-Difluoromethyl-5,4’-dimethoxygenistein (DFMG) on the proliferation and migration of vascular smooth muscle cells(VSMCs)stimulated by LPC-induced injured vascular endothelial cells(VECs), and whether the intervention process of DFMG is related to the TLR4. Methods The proliferation and migration of VSMCs was detected by CCK-8 assay and Transwell-migration method. The TLR4 protein and gene expression of VECs was detected by western blot and fluorescence quantitative PCR. Using TLR4 specific agonist (LPS) and specific inhibitor (CLI095), the prolif-eration and migration ability of VSMCs was analysed by CCK-8 assay and Transwell-migration method. Results The injured VECs induced by LPC caused the proliferation and migration of VSMCs, and VSMCs cocultured with VECs induced by 30 μM LPC became the injured coculture model of the following experiment. TLR4 protein expression of the injured VECs in the in-jured coculture model was 2.3 times as much as that of the normal VECs in the normal coculture model; TLR4 gene expression of the injured VECs in the injured coculture model was 2.4 times as much as that of the normal VECs in the normal coculture model. The proliferation and migration capacity of the injured group and the agonist group were stronger than that of the normal group. In addition, the proliferation and migration capacity of the inhibitor group and the DFMG-treated group were weaker than that of the injured group. Conclusion The injured VECs induced by LPC caused the proliferation and migration of VSMCs, and DFMG inhibited proliferation and migration of VSMCs in the LPC-induced injured VECs-VSMCs cocultured model by de-pressing TLR4.
出处
《湖南师范大学学报（医学版）》
2016年第1期1-5,共5页
Journal of Hunan Normal University(Medical Sciences)
基金
国家自然科学基金资助项目(81370382)
湖南省自然科学基金项目(14JJ2059)
