Konaka Ken, Moriyama Kota, Sakurada Takumi, Okada Naoto, Imanishi Masaki, Zamami Yoshito, Kawazoe Kazuyoshi, Fushitani Shuji, Ishizawa Keisuke
Department of Pharmacy, Tokushima Municipal Hospital, 2-34 Kitajyosanjima, Tokushima, 770-0812 Japan.
Department of Pharmacy, Shimane University Hospital, 89-1 Enya, Izumo, 693-0021 Japan.
J Pharm Health Care Sci. 2017 Jul 21;3:20. doi: 10.1186/s40780-017-0090-y. eCollection 2017.
In chemotherapy, the full round of treatment must be completed as scheduled to achieve the strongest therapeutic effect. However, peripheral neuropathy, a severe side effect of the chemotherapeutic agent paclitaxel, can force the premature discontinuation of treatment. As some kampo practitioners have suggested that it may be possible to counteract such side effects, we analyzed the effects of Kamishoyosan, Shakuyakukanzoto, and Goshajinkigan in an in vitro model of paclitaxel-induced peripheral neuropathy.
Paclitaxel-treated PC12 cells were assessed for neurite length and performed Western blot analysis for growth-associated protein-43 (GAP-43) and light neurofilament protein (NF-L) levels in the presence of nerve growth factor (NGF); they were re-assessed, with additional testing for acetylcholinesterase levels, after application of one of the kampo. We also compared phosphorylation of extracellular signal-regulated kinase (Erk)1/2 and Akt via Western blot analysis. About effect of kampo to anticancer efficacy, we confirmed cell cytotoxicity in A549 cells using MTT assay.
Addition of Kamishoyosan or Shakuyakukanzoto, but not Goshajinkigan, significantly improved neurite length and GAP-43 and NF-L levels from paclitaxel-treated PC12 cells, relative to those of only NGF-treated PC12 cells. The promoting effect of Kamishoyosan and Shakuyakukanzoto in neurite outgrowth is confirmed when NGF promoted neurite outgrowth, and it was inhibited partially when Erk1/2 and Akt were blocked by Erk1/2 inhibitor or Akt inhibitor alone. Furthermore, neurite outgrowth induced by TJ24 and TJ68 was inhibited more strongly when Erk1/2 inhibitor and Akt inhibitor were treated at the same time. NGF with Kamishoyosan or Shakuyakukanzoto promoted the proportion of phosphorylated Erk1/2 and phosphorylated Akt compare with NGF only. On the other hand, Kamishoyosan or Shakuyakukanzoto didn't influence cytotoxicity of paclitaxel in A549 cells.
Kamishoyosan or Shakuyakukanzoto promotes neurite outgrowth with NGF via increasing the proportion of phosphorylated Erk1/2 and phosphorylated Akt in PC12 cells. The effect applies to recovery from paclitaxel-induced axonal involvement and might promote recovery from paclitaxel-induced neuropathy without influence of anticancer effect of paclitaxel.
在化疗中，必须按计划完成整个疗程以达到最强的治疗效果。然而，化疗药物紫杉醇的严重副作用——周围神经病变，可能会迫使治疗提前终止。由于一些汉方从业者认为有可能抵消此类副作用，我们在紫杉醇诱导的周围神经病变的体外模型中分析了加味逍遥散、芍药甘草汤和五积散的效果。
在存在神经生长因子（NGF）的情况下，评估紫杉醇处理的PC12细胞的神经突长度，并对生长相关蛋白43（GAP-43）和轻神经丝蛋白（NF-L）水平进行蛋白质印迹分析；在应用其中一种汉方药物后，再次评估细胞，并额外检测乙酰胆碱酯酶水平。我们还通过蛋白质印迹分析比较了细胞外信号调节激酶（Erk）1/2和Akt的磷酸化情况。关于汉方药物对抗癌疗效的影响，我们使用MTT法在A549细胞中确认了细胞毒性。
与仅用NGF处理的PC12细胞相比，添加加味逍遥散或芍药甘草汤，但不添加五积散，可显著改善紫杉醇处理的PC12细胞的神经突长度以及GAP-43和NF-L水平。当NGF促进神经突生长时，加味逍遥散和芍药甘草汤对神经突生长的促进作用得到证实，而当Erk1/2和Akt单独被Erk1/2抑制剂或Akt抑制剂阻断时，这种促进作用会部分受到抑制。此外，当同时用Erk1/2抑制剂和Akt抑制剂处理时，TJ24和TJ68诱导的神经突生长受到的抑制更强。与仅用NGF相比，NGF与加味逍遥散或芍药甘草汤一起可促进磷酸化Erk1/2和磷酸化Akt的比例。另一方面，加味逍遥散或芍药甘草汤不影响紫杉醇对A549细胞的细胞毒性。
加味逍遥散或芍药甘草汤通过增加PC12细胞中磷酸化Erk1/2和磷酸化Akt的比例，与NGF一起促进神经突生长。这种作用适用于从紫杉醇诱导的轴突损伤中恢复，并且可能促进从紫杉醇诱导的神经病变中恢复，而不影响紫杉醇的抗癌效果。
