人参-三七-川芎提取物对内皮微粒介导HUVECs衰老的干预作用

目的:观察人参-三七-川芎提取物对延缓内皮微粒(EMPs)诱导的血管内皮细胞衰老的影响,并探索其作用机制.方法:以人脐静脉内皮细胞(HUVECs)为研究对象,复制性衰老10～12代细胞作为衰老模型,实验分为年轻组(2～4代细胞)、衰老组(10～ 12代细胞)、单纯EMPs干预组(提取衰老细胞产生的EMPs来干预年轻细胞...

Effect of Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma Extract on HUVECs Senescence Mediated by Endothelial Microparticles

Objective: To investigate the effect of Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract on endothelial microparticles(EMPs)-induced vascular endothelial cell senescence,and explore the possible mechanism. Method: Human umbilical vein endothelial cells(HUVECs)were used as the research objects,and the aged model was established with 10-12 passages of replicative senescence cells. The experimental cells were divided into young group(2-4 passage cells),aged group(10-12 passage cells),only EMPs intervention group(extract EMPs produced by aged cells to intervene young cells)and low dose,middle dose and high dose drug intervention groups(200,300,400 mg·L-1). Senescence relatedβ-galactosidase(SA-β-gal)staining and cell cycle propidium iodide(PI)staining were used to determine cell senescence. Cell counting kit-8(CCK-8)assay was used to screen the drug concentration. EMPs were extracted by two-step centrifugation,EMPs labeled with phycoerythrin(PE)anti-human CD31 antibody or fluorescein isothiocyanate(FITC)annexin V were detected by flow cytometry,intracellular reactive oxygen species(ROS)were detected by 2’,7’-dichlorofluorescein diacetate(DCFDA)staining. Result:After treatment with the drug,SA-β-gal activity of the aged cells significantly decreased(P<0.01),the S phase arrest was restored(P<0.01),and the number of CD31+EMPs and annexin V+EMPs secreted by aged cells decreased(P<0.05). Compared with the young group,only EMPs intervention group could induce increased SA-β-gal activity and S phase arrest in young cells(P<0.05,P<0.01). However,after intervention of EMPs and the drug,EMPs-mediated increase of SA-β-gal activity was significantly inhibited and S phase arrest was restored(P<0.05). The increase of intracellular ROS induced by EMPs was also significantly inhibited by the drug(P<0.05,P<0.01). Conclusion:Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract can delay the senescence of vascular endothelial cells by influencing EMPs, and the mechanism may be related to the inhibition of increased intracellular ROS induced by EMPs.