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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 10μmol/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HBVSMCs DDR2的表达机制与ERK1/2信号转导途径有关。
Objective To investigate the mechanism of peroxynitrite (ONOO-) induced expression of discoid domain receptor 2 (DDR2) in human cerebrovascular smooth muscle cells (HBVSMCs). Methods HBV SMMCs were cultured in vitro. Morphological and cell survival rates of cells pretreated with PD98059 were determined by inverted phase contrast microscope, acridine orange staining and MTT colorimetric assay. Western blot and Real-time PCR were used to detect the effect of ONOO- The expression of DDR2 / MMP-9 in HBV-infectedMCSs and the expression of DDR2 / MMP-9 in ERK1 / 2 inhibitor PD98059. Results The expression of DDR2 at 10μmol / L ONOO- induced high levels of protein and mRNA (P <0.05). With the increase of ONOO- concentration, DDR2 gradually decreased (P <0.05). There was no obvious morphological change of HBV pre-treated HBV-SMMCs, and no significant effect was found on the survival rate of HBV-infected MCs (P> 0.05). PD98059 significantly inhibited ONOO-induced DDR2 / MMP-9 expression (P <0.05). Conclusion ERK1 / 2 signal pathway inhibitor PD98059 can inhibit ONOO-induced DDR2 / MMP-9 expression. The mechanism of ONOO-induced DDR2 expression in HBV SMMC-memory cells is related to ERK1 / 2 signal transduction pathway.