Oshima Y, Sakamoto T, Kawano Y, Hata Y, Yoshikawa H, Sonoda K H, Ishibashi T, Inomata H
Department of Ophthalmology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Graefes Arch Clin Exp Ophthalmol. 1998 Jan;236(1):52-60. doi: 10.1007/s004170050042.
The combined effect of electric pulses (EP) and antiproliferative agents on the proliferation of rabbit Tenon's capsule fibroblasts was investigated.
Rabbit Tenon's capsule fibroblasts were cultured. Some of these cells were exposed to various intensities of EP alone (500-2500 V/cm). Other cells were then exposed for 30 min to an antiproliferative agent: bleomycin (BLM; 0.0005-50 mumol/l), mitomycin C (MMC; 0.0005-50 mumol/l), 5-fluorouracil (5-FU; 0.05-5000 mumol/l), or streptomycin (SM; 0.0005-50 mumol/l) with or without EP (2000 V/cm, 99 mus, eight pulses). Cell proliferation was assessed by cell counting on day 3 and by a 3H-thymidine uptake assay. DNA fragmentation was assessed by flow-cytometric analysis and agarose gel electrophoresis.
A significant reduction in the cell number was observed only at 2500 V/cm (P < 0.05). BLM, MMC and 5-FU treatment inhibited cell proliferation in a dose-dependent manner either with or without EP (ID50: BLM alone, 0.029 mumol/l; BLM and EP, 0.00022 mumol/l; MMC alone, 41.6 mumol/l; MMC and EP, 27.5 mumol/l; 5-FU alone, 1045 mumol/l; 5-FU and EP, 690.2 mumol/l; P < 0.05). EP treatment induced an inhibitory effect of BLM on cell proliferation which was 100 times more prominent than BLM alone (0.0005 mumol/l of BLM alone 103.4 +/- 4.4%, 0.0005 mumol/l of BLM and EP 26.0 +/- 4.4%; P = 0.021). BLM treatment with EP also augmented the apoptotic-like DNA fragmentation in both a flow-cytometric DNA histogram and agarose gel-electrophoresis.
EP treatment enhanced the inhibitory effect of BLM on the cell proliferation of Tenon's capsule fibroblasts of rabbits. The combination of electric pulses and antiproliferative drug treatments may therefore reduce the necessary dose of antiproliferative agents in filtering surgery.
研究电脉冲（EP）与抗增殖剂联合作用对兔眼Tenon囊成纤维细胞增殖的影响。
培养兔眼Tenon囊成纤维细胞。部分细胞单独暴露于不同强度的电脉冲（500 - 2500 V/cm）。其他细胞随后暴露于抗增殖剂30分钟：博来霉素（BLM；0.0005 - 50 μmol/l）、丝裂霉素C（MMC；0.0005 - 50 μmol/l）、5 - 氟尿嘧啶（5 - FU；0.05 - 5000 μmol/l）或链霉素（SM；0.0005 - 50 μmol/l），有或无电脉冲（2000 V/cm，99 μs，8个脉冲）。在第3天通过细胞计数和3H - 胸腺嘧啶核苷摄取试验评估细胞增殖。通过流式细胞术分析和琼脂糖凝胶电泳评估DNA片段化。
仅在2500 V/cm时观察到细胞数量显著减少（P < 0.05）。BLM、MMC和5 - FU处理无论有无电脉冲均以剂量依赖性方式抑制细胞增殖（半数抑制浓度：单独BLM为0.029 μmol/l；BLM与电脉冲联合为0.00022 μmol/l；单独MMC为41.6 μmol/l；MMC与电脉冲联合为27.5 μmol/l；单独5 - FU为1045 μmol/l；5 - FU与电脉冲联合为690.2 μmol/l；P < 0.05）。电脉冲处理增强了BLM对细胞增殖的抑制作用，比单独使用BLM时显著突出100倍（单独0.0005 μmol/l的BLM为103.4 ± 4.4%，0.0005 μmol/l的BLM与电脉冲联合为26.0 ± 4.4%；P = 0.021）。BLM与电脉冲联合处理在流式细胞术DNA直方图和琼脂糖凝胶电泳中也增强了凋亡样DNA片段化。
电脉冲处理增强了BLM对兔眼Tenon囊成纤维细胞增殖的抑制作用。因此，电脉冲与抗增殖药物联合处理可能会减少滤过性手术中抗增殖剂的必要剂量。
