PD98059抑制Peroxynitrite诱导人脑血管平滑肌细胞DDR2的表达

目的 探讨过氧亚硝酸盐(ONOO-)诱导人脑血管平滑肌细胞(HBVSMCs)盘状结构域受体2(DDR2)表达的机制.方法 体外培养HBVSMCs,用倒置相差显微镜、吖啶橙染色法和MTT比色法对PD98059预处理的细胞进行细胞形态学及细胞生存率检测,采用Western blot及Real-time PCR技术,检测ONOO-作用下HBVSMCs中DDR2表达及施加ERK1/2抑制剂PD98059后DDR2/MMP-9表达的变化.结果 10 μmol/L ONOO-诱导DDR2在蛋白及mRNA水平上呈现高表达(P＜0.05),随着ONOO-浓度的增加,DDR2逐渐呈现低表达(P＜0.05).PD98059预处理后的HBVSMCs,无明显形态学变化,对其进行生存率检测亦无显著影响(P＞0.05).PD98059显著抑制ONOO-诱导的DDR2/MMP-9表达(P＜0.05).结论 ERK1/2信号转导途径抑制剂PD98059可以抑制ONOO-诱导的DDR2/MMP-9表达,ONOO-诱导HBVSMCsDDR2的表达机制与ERK1/2信号转导途径有关.

PD98059 inhibits Peroxynitrite-induced discoidin domain receptor-2 expression in human brain vascular smooth muscle cells

Objective To explore the possible mechanism underlying peroxynitrite(ONOO-)-induced discoidin domain receptor-2(DDR2) expression in human brain vascular smooth muscle cells(HBVSMCs).Methods HBVSMCs were cultured in vitro in different concentrations of ONOO-,and PD98059,a specific inhibitor of ERK1/2.Morphology and viability of HBVSMCs pretreated with PD98059 were detected by an inverted phase contrast microscope,acridine orange staining and MTT assay.Western blot and Real-time PCR were employed to determine expressions of DDR2/ MMP-9.Results 10 μmol/L of ONOO-significantly induced expression of DDR2 at both protein and mRNA levels(P<0.05).With increase in concentrations of ONOO-,expression of DDR2 was markedly decreased(P<0.05).After pretreatment on HBVSMCs with PD98059,cellular morphology and viability had no significant change by an inverted phase contrast microscope,acridine orange staining and MTT assay(P>0.05).Pretreatment on HBVSMCs with PD98059 significantly inhibited expressions of DDR2/ MMP-9 in HBVSMCs induced by 10 μmol/L of ONOO-(P<0.05).Conclusion PD98059 inhibits ONOO——induced DDR2 expression.The ERK1/2 signal transduction pathway might involve in ONOO——induced DDR2 expression in HBVSMCs.