PDGFR-β反义寡核苷酸对大鼠血管平滑肌细胞凋亡的影响

Effects of PDGFR-β antisense oligonucleotide on apoptosis of rat vascular smooth muscle cells

目的 :探讨血小板衍化生长因子 β受体 (PDGFR- β)反义寡核苷酸对培养的大鼠血管平滑肌细胞 (VSMC)凋亡的影响。方法 :建立 SD大鼠胸主动脉 VSMC体外增殖模型 ,取 5～ 10代细胞为实验对象 ,按实验目的分为以下几组 :1反义寡核苷酸组 ;2正义寡核苷酸组 ;3错义寡核苷酸 ;4空白对照组。利用透射电镜等观察 VSMC加药前后形态学和超微结构的变化 ,采用原位末端脱氧核苷酸转移酶介导的 d U TP缺口末端标记法 (TU NEL)和流式细胞仪观察和分析 PDGFR- β反义寡核苷酸对 VSMC凋亡的影响。结果 :1加药后 2 4h细胞形态和超微结构较加药前无明显改变 ,48h细胞形态和超微结构较加药前有明显改变。2加药后 2 4h TU NEL 染色各组未发现凋亡细胞 ,48h后反义寡核苷酸组细胞爬片染色有较多凋亡阳性细胞出现。 3加药 2 4h流式细胞仪检测未发现加药组细胞凋亡量与对照组之间有显著差异 ,48h加药组细胞凋亡量明显高于对照组。结论 :PDGFR- β反义寡核苷酸对大鼠VSMC凋亡有诱导作用。

AIM: To study the effects of PDGFR-β antisense oligonucleotides on apoptosis of cultured rat vascular smooth muscle cells(VSMC). METHODS: Cultured rat VSMC proliferation model was established in vitro,cells of 5～10 passages were used for the experiment. VSMC were divided into four groups:①control,② PDGFR-β antisense oligonucleotide,③ PDGFR-β sense oligonucleotide,④ PDGFR-β scrambled oligonucleotide. Phase contrast microscope and electron microscope were used to observe the change of cell morphology and cell ultrastructure. Apoptotic VSMC were detected and assayed by terminal deoxynucleotidyl transferase-mediated dUTP nick-enddd labeled(TUNEL) and flow cytometry. RESULTS: ①No difference of morphology and ultrastructure was observed between control group and other groups at 24 h. ② Compared with control group,cell morphology and ultrastructure of AODN group varied at 48 h;No apoptotic cell was detected by TUNEL at 24 h. Many apoptotic cells were detected at 48 h by TUNEL in the AODN group.③ There were no difference of apoptosis between control group and AODN group at 24 h. Compared with control group, the numble of apoptotic cells of AODN group was obviously higher at 48 h. CONCLUSION: These results suggest that PDGFR-β AODN can induce the apoptosis of cultured VSMC.