NO影响CAP化疗损伤效应及其机制研究

目的 探讨一氧化氮是否提高CAP对L1210细胞损伤效应及其机制研究。方法 将L1210细胞加入隔离培养器内与3T3细胞共培养，收集各组L1210细胞12、24、48、72h的标本，台盼蓝染色观察直接损伤的作用，TUNNEL法观察诱导细胞凋亡的情况，流式细胞检测细胞周期观察细胞的增殖状态。结果 质粒转染组L1210细胞较空载体组台盼蓝拒染率下降更快，12h开始两组差异有统计学意义（P〈0．05）。DEVD-CHO在24h后能够提高台盼蓝拒染率（P〈0．05）。质粒转染组的L1210细胞凋亡率升高更快，与空载体转染组各时相点有显著差异（P〈0．05）。DEVD-CHO能够使细胞凋亡率下降（P〈0．05）。CAP作用4h后各组L1210细胞周期G1期显著升高，S期迅速下降，质粒转染组比空载体转染组G1期升高快，加入DEVD-CHO4h后G1期上升较单纯质粒转染组慢（P〈0．05）。结论 NO可增强CAP对L1210细胞的直接损伤作用：促进细胞凋亡，使CAP对细胞G1期的阻滞作用更敏感、持续时间更长。上述作用也与Caspase-3的活化有关。

Damaging effect of No on cyclophosphamide and its mechanism

Objective To study whether NO could enhance the damaging effect of cyclophosphamide(CAP) on L1210 cells were in separated culture condition.Metheds L1210 cells were co-cultured with different transfected 3T3 cells.The direct effects of NO and NO-induced apoptosis of L1210 cells in different groups at 12,24,48,72h were detected by trypan blue rejecting dyeing(TRD) and TUNNEL,respectively.The cell cycles were detected by flow cytometry.Results The rate of TRD of L1210 cells was significantly lower in iPT group than that in PT group during 12-72 h after CAP treatment(P<0.05).The apoptosis of L1210 cells in iPT group after CAP treatment was significantly higher than that in PT group during 12-72 h(P<0.05).There was also significant difference between iPT group and iPT plus DEVD-CHO group(P<0.05).L1210 cells at G1 phase increased more quickly at 4h after CAP treatment in iPT group.L1210 cells at S phase declined more quickly at 4h in iPT group after CAP treatment.In iPT plus DEVD-CHO group,L1210 cells at G1 phase increased more slowly at 4h.Conclusion NO can enhance the damaging effect of CAP on L1210 cells through direct damage to L1210 cells,enhancement of apoptosis,and blockage of G1 phase,which are dependent on Caspase-3 mechanism.