Konings P N, Makkink W K, van Delft A M, Ruigt G S
Department of Neuropharmacology, Organon International BV, Oss, The Netherlands.
Brain Res. 1994 Mar 21;640(1-2):195-204. doi: 10.1016/0006-8993(94)91873-2.
Cytostatic drugs, like cisplatin, vincristine and taxol, when given to cancer patients may cause peripheral neuropathies. We were interested in the potential neuroprotective effects of neurotrophic factors against such neuropathies. To this aim we studied the effects of these cytostatic agents on sensory fibers located in the dorsal root ganglia (DRG) in vitro and studied whether nerve growth factor (NGF) could reverse the cytostatic induced morphological changes on intact DRG (1 DRG/well, n = 10 per dose). Neuritogenesis from DRG was measured with an image analysis system following exposure to different concentrations of cytostatic drugs in the presence of 3 ng NGF/ml and cytosine arabinoside (Ara-C, 10(-6) M). Relative neurite outgrowth in intact DRG in culture was reduced dose-dependently, (a) by vincristine starting at a dose of 0.4 ng/ml for 2 days (-33% as compared to control; P < 0.001, Student's t-test); (b) by taxol 10 ng/ml (-60%; P < 0.001), and (c) by cisplatin 3 micrograms/ml (-47%, P < 0.001). Cisplatin also prevented the migration of satellite cells away from the intact DRG along the extending neurites into the well in contrast to control, vincristine, or taxol. To evaluate the neuroprotective potential of NGF in this in vitro cytostatic neuropathy model, we incubated intact DRG with cytostatic agents in combination with increasing amounts of NGF. Neurite outgrowth from DRG treated with vincristine (0.5 ng/ml)+NGF (3 ng/ml) for 2 days was significantly higher (+87%) than after treatment with vincristine + 1 ng NGF/ml (P < 0.001). Neurite outgrowth from DRG treated with taxol (20 ng/ml)+NGF (3 ng/ml) for 2 days was significantly higher (+228%) than after taxol + 1 ng NGF/ml (P < 0.05). Neuritogenesis from DRG treated with cisplatin (2.5 micrograms/ml)+NGF (3 ng/ml) for 2 days was significantly increased (+105%) compared to treatment with cisplatin + 1 ng NGF/ml (P < 0.001). DRG thus appear to be a very suitable model for studying cytostatic drug-induced neuropathies in vitro and NGF has a clear neuroprotective effect on the vincristine-, taxol-, and cisplatin-induced neuropathies in this in vitro model.
细胞毒性药物，如顺铂、长春新碱和紫杉醇，在给予癌症患者时可能会导致周围神经病变。我们对神经营养因子针对此类神经病变的潜在神经保护作用感兴趣。为此，我们在体外研究了这些细胞毒性药物对背根神经节（DRG）中感觉纤维的影响，并研究了神经生长因子（NGF）是否能逆转细胞毒性药物对完整DRG（每孔1个DRG，每个剂量n = 10）诱导的形态学变化。在存在3 ng NGF/ml和阿糖胞苷（Ara-C，10(-6) M）的情况下，将DRG暴露于不同浓度的细胞毒性药物后，用图像分析系统测量DRG的神经突生成。培养的完整DRG中的相对神经突生长呈剂量依赖性降低，（a）长春新碱从0.4 ng/ml的剂量开始作用2天（与对照组相比降低33%；P < 0.001，学生t检验）；（b）紫杉醇10 ng/ml（降低60%；P < 0.001），以及（c）顺铂3微克/ml（降低47%，P < 0.001）。与对照组、长春新碱或紫杉醇相比，顺铂还阻止了卫星细胞沿延伸的神经突从完整的DRG迁移到孔中。为了评估在这个体外细胞毒性神经病变模型中NGF的神经保护潜力，我们将完整的DRG与细胞毒性药物以及增加量的NGF一起孵育。用长春新碱（0.5 ng/ml）+ NGF（3 ng/ml）处理2天的DRG的神经突生长明显高于用长春新碱 + 1 ng NGF/ml处理后（增加87%；P < 0.001）。用紫杉醇（20 ng/ml）+ NGF（3 ng/ml）处理2天的DRG的神经突生长明显高于用紫杉醇 + 1 ng NGF/ml处理后（增加228%；P < 0.05）。用顺铂（2.5微克/ml）+ NGF（3 ng/ml）处理2天的DRG的神经突生成与用顺铂 + 1 ng NGF/ml处理相比明显增加（增加105%；P < 0.001）。因此，DRG似乎是体外研究细胞毒性药物诱导的神经病变非常合适的模型，并且在这个体外模型中，NGF对长春新碱、紫杉醇和顺铂诱导的神经病变具有明显的神经保护作用。
