DDR2在Peroxynitrite诱导人中枢血管平滑肌细胞凋亡中的表达

Expression of DDR2 in human cerebral vascular smooth muscle cells apoptosis subjected to peroxynitrite

目的探究过氧亚硝酸盐(peroxynitrite,ONOO-)诱导人中枢血管平滑肌细胞(HCVSMCs)凋亡过程中盘状结构域受体(DDR2)基因表达的变化。方法以不同浓度的ONOO-作用于体外培养的HCVSMCs,采用噻唑蓝(MTT)比色法测定细胞生存率,流式细胞仪测定细胞凋亡率,吖啶橙染色进行形态学观察。同时,采用逆转录-聚合酶链反应(RT-PCR)及Western blot检测DDR2mRNA及蛋白表达。结果与对照组相比,以ONOO-作用于HCVSMCs24h可显著抑制该系细胞增生,诱导细胞凋亡(P<0.05),并引起DDR2mRNA及蛋白水平总体呈现下调趋势(P<0.05)。结论 DDR2基因可能在ONOO-诱导HCVSMCs凋亡的信号转导调节通路中起关键作用。

Objective To investigate expression of the DDR2 gene in human cerebral vascular smooth muscle cells （HCVSMCs） exposed to peroxynitrite （ONOO-）. Methods HCVSMCs cultured in vitro were subjected to different concentrations of ONOO-. Cell viability was measured by MTT assay. The rate of cell apoptosis was measured by a flow cytometer. The morphological changes were assessed by acridine orange staining to determine whether cells experienced apoptosis under the stress of ONOO-. Expression of the DDR2 gene at both mRNA and protein levels was simultaneously examined by the means of RT-PCR and Western-blot. Results Direct exposure of cells to ONOO-was able to block cell proliferation in a concentration-dependent manner via the apoptotic pathway. Under such stress,expression of the DDR2 gene was finally decreased at both mRNA and protein levels. Conclusion The DDR2 gene may play a pivotal role in ONOO--induced apoptosis in HCVSMCs.