Histological, cellular and behavioural analyses of effects of chemotherapeutic agent cyclophosphamide in the developing cerebellum
Histological, cellular and behavioural analyses of effects of chemotherapeutic agent cyclophosphamide in the developing cerebellum
复制标题
化疗药物环磷酰胺对小脑发育影响的组织学、细胞和行为分析
DOI：
10.1111/cpr.12608
复制
发表时间：
2019
影响因子：
8.5
通讯作者：
Gao Wei-Qiang
中科院分区：
文献类型：
--
作者：
Zhang Yu;Li Yongfang;Luo Wenqin;Tang Yaohui;Wang Jia;Yang Ru;Gao Wei-Qiang
本研究采用C57 BL/6小鼠和Math 1依赖性GFP表达转基因小鼠，通过组织学、细胞学和行为学研究，探讨环磷酰胺（cyclophosphamide，CTX）对儿童小脑发育的影响，为CTX在儿童小脑发育中的临床应用提供参考。H&E染色分析CTX对小脑的组织学影响。EdU染色和TUNEL法检测细胞增殖和凋亡。采用转棒试验和悬丝试验评价大鼠的行为功能。免疫荧光染色鉴定细胞类型。通过定量真实的时间PCR或immunoblotting.ResultsWe发现，虽然CTX诱导细胞增殖显着减少，并增加在EGL的凋亡在48小时内，行为功能和小脑的多层层状结构，在很大程度上恢复时，小鼠生长到成年人的分化标志物和基因的测定。机制上，颗粒神经元祖细胞在SHH信号的驱动下，在CTX给药停止后迅速增强增殖能力，这使得发育中的小脑能够赶上并逐渐补充损伤。但如果在使用后不久停止使用该药物，小脑多层层状结构和运动功能可以在很大程度上恢复。
ObjectivesWe performed histological, cellular and behavioural analyses of the effects of cyclophosphamide (CTX), a chemotherapeutic drug, in the developing cerebellum and aimed to provide valuable insights into clinical application of CTX in children.Materials and methodsC57BL/6 mice and Math1‐dependent GFP expression transgenic mice were used in the research. H&E staining was performed to analyse histological effects of CTX in the cerebellum. Staining for EdU and TUNEL was used to estimate the cell proliferation and apoptosis. Rotarod test and hanging wire test were used to evaluate the behavioural functions. Immunofluorescent staining was used to identify the cell types. The differentiation markers and genes related to Sonic Hedgehog (SHH) signalling were measured via quantitative real‐time PCR or immunoblotting.ResultsWe found that while CTX induced a significant reduction in cell proliferation and increased apoptosis in the EGL in 48 hours, the behavioural functions and the multilayer laminar structure of cerebella were largely restored when the mice grew to adults. Mechanistically, granule neuron progenitors, driven by the SHH signalling, enhanced the capability of proliferation quickly after CTX administration was stopped, which allowed the developing cerebellum to catch up and to gradually replenish the injury.ConclusionThe chemotherapeutic agent CTX induces an immediate damage to the developing cerebellum, but the cerebellar multilayer laminar structure and motor function can be largely restored if the agent is stopped shortly after use.