摘要
目的探讨人参皂苷Re通过调控Ras同源基因家族成员A(Ras homologous gene family member A,RhoA)/丝裂原活化蛋白激酶(Mitogen activated protein kinase,MAPK)通路对血管紧张素Ⅱ(AngiotensinⅡ,AngⅡ)诱导的血管平滑肌细胞(Vascular smooth muscle cells,VSMCs)增殖、迁移和表型转化的影响。方法MOVAS细胞分为对照组、AngⅡ组、人参皂苷Re低剂量组、人参皂苷Re中剂量组、人参皂苷Re高剂量组、人参皂苷Re高剂量+RhoA激活剂(U46619)组。采用CCK-8检测MOVAS细胞增殖;划痕实验检测MOVAS细胞迁移;免疫细胞化学法检测MOVAS细胞中α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、骨桥蛋白(Osteopontin,OPN)蛋白阳性表达;qRT-PCR检测MOVAS细胞PCNA、MMP-9、MMP-2 mRNA表达;Western blot检测MOVAS细胞中RhoA、磷酸化细胞外信号调节激酶1/2(phosphorylated extracellularsignalregulatedkinase1/2,p-ERK1/2)、磷酸化应激活化蛋白激酶1(phosphorylatedstress activated protein kinase 1,p-JNK1)、p-P38蛋白。结果与对照组比较,AngⅡ组MOVAS细胞OD450值、划痕愈合率、OPN蛋白阳性表达、PCNA、MMP-9、MMP-2 mRNA表达及RhoA、p-ERK1/2、p-JNK1、p-P38蛋白升高,α-SMA蛋白阳性表达显著降低(P<0.05);与AngⅡ组比较,人参皂苷Re低、中、高剂量组MOVAS细胞OD450值、划痕愈合率、OPN蛋白阳性表达、PCNA、MMP-9、MMP-2 mRNA表达及RhoA、p-ERK1/2、p-JNK1、p-P38蛋白降低,α-SMA蛋白阳性表达升高,且人参皂苷Re高剂量组趋势最显著(P<0.05);与人参皂苷Re高剂量组比较,人参皂苷Re高剂量+U46619组MOVAS细胞OD450值、划痕愈合率、OPN蛋白阳性表达、PCNA、MMP-9、MMP-2 mRNA表达及RhoA、p-ERK1/2、p-JNK1、p-P38蛋白升高,α-SMA蛋白阳性表达显著降低(P<0.05)。结论人参皂苷Re抑制AngⅡ的MOVAS细胞增殖、迁移和表型转化可能与抑制RhoA/MAPK通路有关。
Objective To investigate the effect of ginsenoside Re on angiotensinⅡ(AngⅡ)-induced proliferation,migration,and phenotype transformation of vascular smooth muscle cells(VSMCs)by regulating the Ras homologous gene family member A(RhoA)/mitogen activated protein kinase(MAPK)pathway.Methods MOVAS cells were divided into control group,AngⅡgroup,ginsenoside Re low dose group,ginsenoside Re medium dose group,ginsenoside Re high dose group,andginsenoside Re high dose+RhoA activator(U46619)group.MOVAS cell proliferation was detected by CCK-8.MOVAS cell migration was detected by scratch assay.Immunocytochemistry was used to detect the positive expression ofα-smooth muscle actin(α-SMA)and osteopontin(OPN)proteins in MOVAS cells.qRT-PCR was used to detect the mRNA expression of PCNA,MMP-9,and MMP-2 in MOVAS cells.Western blot was used to detect RhoA,phosphorylated extracellular signal regulated kinase 1/2(p-ERK1/2),phosphorylated stress activated protein kinase 1(p-JNK1),and p-P38 protein in MOVAS cells.Results Compared with the control group,the OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression,RhoA,p-ERK1/2,p-JNK1,p-P38 protein expression of MOVAS cells in the Ang II group increased,while the positive expression ofα-SMA protein decreased significantly(P<0.05).Compared with the AngⅡgroup,the OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression,and RhoA,p-ERK1/2,p-JNK1,and p-P38 proteins of MOVAS cells in the low,medium,and high dose groups of ginsenoside Re decreased,while the positive expression ofα-SMA protein increased.The trend was most significant in the high dose group of ginsenoside Re(P<0.05).Compared with ginsenoside Re high-dose group,OD450 value,scratch healing rate,OPN protein positive expression,PCNA,MMP-9,MMP-2 mRNA expression and RhoA,p-ERK1/2,p-JNK1,p-P38 protein in ginsenoside Re high-dose+U46619 group increased,the positive expression ofα-SMA protein decreased significantly(P<0.05).Conclusion Ginsenoside Re may inhibit the proliferation,migration,and phenotype transformation of AngⅡin MOVAS cells by suppressing the RhoA/MAPK pathway.
作者
胡威威
王晓玲
李晓荣
田春辉
刘智芬
HU Weiwei;WANG Xiaoling;LI Xiaorong;TIAN Chunhui;LIU Zhifen(Hengshui people’s hospital,Hengshui 053000,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第5期1417-1425,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
衡水市科学技术局科学技术研究与发展计划项目(2021014089Z):替米沙坦联合羟苯磺酸钙对高血压并糖尿病血压节律研究,负责人:胡威威。
关键词
人参皂苷RE
血管紧张素Ⅱ
血管平滑肌细胞
高血压
增殖
表型转化
Ginsenoside Re
AngiotensinⅡ
Vascular smooth muscle cells
Hypertension
Proliferation
Phenotypic transformation
