【摘要】 目的 :研究反义c myc寡核苷酸抑制大鼠气道平滑肌增殖与细胞凋亡的关系 .方法 :大鼠气道平滑肌细胞原代培养 ,4～ 12代用于实验 .利用Lipofectin将正义、反义及错配c myc寡核苷酸导入大鼠气道平滑肌细胞 ,姬姆萨染色查找凋亡小体 ,流式细胞技术观察有无凋亡峰出现 ,琼脂糖电泳观察凋亡细胞特有的梯状图谱 ,免疫组织化学染色检测凋亡相关基因bax和bcl 2 .结果 :反义c myc寡核苷酸处理大鼠气道平滑肌细胞 2 4h后 ,细胞生长明显受到抑制 (0 .12± 0 .0 3vs 0 .19± 0 .0 5 ,P <0 .0 1) ,但无凋亡小体发现 ,流式细胞技术没有检测到凋亡峰 ,细胞总DNA琼脂糖电泳没有观察到典型的梯状凋亡图谱 ,免疫组织化学染色显示凋亡相关基因产物Bax及Bcl 2在各组表达无明显差异 .结论 :反义c myc寡核苷酸抑制大鼠气道平滑肌增殖与细胞凋亡无关更多还原

【Abstract】 AIM: To study the relationship between antiproliferative effect of c myc antisense oligonucleotide in inhibiting rat airway smooth muscle cells proliferation and cell apoptosis. METHODS: Rat airway smooth muscle cells were primary cultured. 4 12 passages were used in experiments. Lipofectin was used to transfect antisense oligonucleotide, sense oligonucleotide and mismatched oligonucleotide for c myc to rat airway smooth muscle cells. Apoptosis bodies were examined after Giemsa’s staining. Flow cytometry technique was used to detect apoptosis peak. DNA ladder was observed after total DNA agarose electrophoresis. The expression of apoptosis associated gene productions bax and bcl 2 were detected by immunohistochemistry staining. RESULTS: Antisense oligonucleotide for c myc could significantly inhibit rat airway smooth muscle cells proliferation 24 h after the treatment(0.12±0.03 vs 0.19±0.05, P < 0.01). No apoptosis body was found. No apoptosis peak was detected by flow cytometry. DNA ladder was not found after total DNA agarose electrophoresis. There was no difference in the expressions of apoptosis associated gene products Bax and Bcl 2 between different groups. CONCLUSION: There is no relationship between antiproliferative effect of c myc antisense oligonucleotide in rat airway smooth muscle cells and cell apoptosis.更多还原