摘要
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.
作者
朱凌波
龚心琰
龚剑萍
倪市毛
宣云岗
ZHU Lingbo;GONG Xinyan;GONG Jianping(Department of Cardiovascular Medicine,Yiwu Central Hospital,Yiwu 322000,China)
出处
《全科医学临床与教育》
2022年第3期206-210,F0003,F0002,共7页
Clinical Education of General Practice
基金
义乌市科技计划项目(19-3-117)
金华市科技计划项目(2019-4-136)。
关键词
自组装短肽
氧化低密度脂蛋白
人脐静脉内皮细胞
血管内皮生长因子
信号通路
self-assembling peptide
oxidized low density lipoprotein
human umbilical vein endothelial cells
vascular endothelial growth factor
notch signaling pathway