丁苯酞修饰新型自组装短肽对氧化低密度脂蛋白损伤血管内皮细胞的影响

Effect of novel self-assembled peptide modified by butylphthalide on ox-LDL-induced vascular endothelial cell injury

目的研究丁苯酞修饰新型自组装短肽(NBP-SAP)对氧化低密度脂蛋白(ox-LDL)损伤血管内皮细胞的影响及与信号通路关系。方法体外培养人脐静脉内皮细胞(HUVECs)并分为NBP-SAP+LY组、NBP-SAP组、ox-LDL组和对照组,分别给予NBP-SAP+LY450139+ox-LDL、NBP-SAP+ox-LDL、ox-LDL及对照处理。Realtime-qPCR法检测不同措施处理后HUVECs中血管内皮生长因子(VEGF)/血管内皮生长因子受体-2(VEGFR-2)-Notch1/DLL4通路相关基因表达,Western blot法检测VEGF/VEGFR2-Notch1、DLL4通路相关蛋白表达量,CCK-8法、Annexin-V/PI双染法、Transwell法分别检测各组细胞活力、细胞凋亡率及细胞侵袭力等。结果NBP-SAP组较ox-LDL组48 h细胞活力及细胞侵袭力增强,细胞凋亡率下降(t分别=3.59、2.91、2.44,P均<0.05),VEGFR-2、B淋巴细胞瘤-2(Bcl-2)蛋白表达量上调(t分别=12.57、14.49,P均<0.05),Notch1、Bax蛋白表达量和DLL4、Notch1、Bax基因mRNA表达下调(t分别=11.30、12.41、13.08、13.16、6.96,P均<0.05)。NBP-SAP+LY组较NBP-SAP组48 h时改善损伤细胞活力、侵袭力的作用减弱,细胞凋亡率升高(t分别=4.38、3.92、4.07,P均<0.05),Notch1、Bax的蛋白表达量上调(t分别=8.59、20.18,P均<0.05),VEGFR-2、Bcl-2蛋白表达量和VEGF、VEGFR-2、Bcl-2基因mRNA表达下调(t分别=14.18、11.23、12.96、19.93、20.33,P均<0.05)。结论NBP-SAP能够激活VEGF/VEGFR2-Notch1/DLL4信号通路,减轻ox-LDL损伤血管内皮细胞的作用。

Objective To evaluate the role of novel self-assembled peptide modified by butylphthalide(NBP-SAP)in the injury of vascular endothelial cells induced by oxidized low density lipoprotein(ox-LDL),and its relationship with vascular endothelial growth factor(VEGF)/Notch1 signaling pathway Methods Human umbilical vein endothelial cells(HUVECs)were divided into NBP-SAP+LY group,NBP-SAP group,ox-LDL group and control group and treated with NBP-SAP plus LY450139 plus ox-LDL,NBP-SAP plus ox-LDL,ox-LDL,phosphate buffer solution.Real-time-qPCR method was used to detect the expression of genes related to VEGF/Notch1 pathway in HUVECs treated with different and Western blot method were used to detect the expression of proteins related to VEGF/Notch1 pathway,CCK-8 method,Annexin-V/PI double staining method and Transwell method were used to detect the cell viability,apoptosis rate and cell invasion and migration of each group.Results Compared to the ox-LDL group,NBP-SAP treatment can improve the damage caused by ox-LDL,enhance the cell vitality and migration,while decrease the apoptosis rate at 48 h(t=3.59,2.91,2.44,P<0.05).The protein expression of VEGFR-2 and Bcl-2 in cells were also increased(t=12.57,14.49,P<0.05).The protein expression of Notch1 and Bax and the mRNA levels of DLL4,Notch1 and Bax genes was down-regulated(t=11.30,12.41,13.08,13.16,6.96,P<0.05).Compared with the NBP-SAP group at 48 h,the NBP-SAP+LY group weakened the effect of improving the viability and invasiveness of injured cells,and the apoptosis rate increased(t=4.38,3.92,4.07,P<0.05),the protein expression of Notch1,Bax increased(t=8.59,20.18,P<0.05),and the protein expression of VEGFR-2,Bcl-2 were up-regulated and the mRNA levels of VEGFR-2,VEGF,Bcl-2 were down-regulated at 48 h(t=14.18,11.23,12.96,19.93,20.33,P<0.05).Conclusion NBP-SAP can alleviate the injury of vascular endothelial cells induced by ox-LDL by activating VEGF/VEGFR2-Notch1/DLL4 pathway.