论文部分内容阅读
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 the changes of DDR2 gene expression during peroxynitrite (ONOO-) induced apoptosis in human central vascular smooth muscle cells (HCVSMCs). Methods Different concentrations of ONOO- were used to in vitro cultured HCVSMCs. MTT assay was used to determine the cell viability. Flow cytometry was used to determine the apoptosis rate. Acridine orange staining was used to observe the morphological changes. At the same time, DDR2 mRNA and protein expression were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Results Compared with the control group, ONOO- induced a significant decrease in cell proliferation and induction of apoptosis (P <0.05) and caused a general downward trend in the expression of DDR2 mRNA and protein (P <0.05). Conclusion The DDR2 gene may play a key role in signal transduction pathway of ONOO-induced apoptosis in HCVSMCs.