Melatonin attenuates nicotine-induced autophagy and neurological changes by decreasing the production of reactive oxygen species
Melatonin attenuates nicotine-induced autophagy and neurological changes by decreasing the production of reactive oxygen species
复制标题
褪黑激素通过减少活性氧的产生来减弱尼古丁诱导的自噬和神经系统变化
DOI：
10.1080/00207454.2019.1692833
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发表时间：
2019
影响因子：
2.2
通讯作者：
Yue
中科院分区：
文献类型：
--
作者：
Yue;Fang Xu;Songmei Wang;Shao;Xuehui Zhang;Yan;Ya;Du;Jianzhong Yin;Yue
摘要目的：本研究的目的是探讨尼古丁毒性的神经学变化机制。材料和方法：采用大鼠嗜铬细胞瘤12（PC 12）细胞和人神经胶质细胞（HM）细胞。用FACScan检测细胞内ROS水平。通过串联单体RFP-GFP标记的LC 3慢病毒监测自噬通量。Western blot检测自噬蛋白LC 3、SQSTM 1/p62和Beclin 1的表达。为探讨尼古丁和褪黑素对神经元形态学的影响，取原代皮层神经元，进行TUBB 3微管蛋白免疫细胞化学染色。结果：尼古丁浓度依赖性地增加PC 12和HM细胞活性氧（ROS）水平。显微镜检查显示尼古丁处理的PC 12细胞自噬通量增加。随后的蛋白质印迹结果显示，尼古丁诱导LC 3B-II和Beclin 1水平升高，并以浓度依赖性方式降低SQSTM 1/p62。最后，尼古丁处理减少了TUBB 3阳性轴突和树突的长度。褪黑素是一种靶向抗氧化剂，可降低ROS水平，阻断尼古丁诱导的自噬激活和形态结构变化。结论：我们的研究结果表明，尼古丁在神经细胞毒性中的作用可能是通过诱导ROS和随后的自噬激活。这些作用可以通过褪黑激素恢复。
Abstract Purpose: The aim of this study was to explore the mechanism of neurological changes underlying the toxicity of nicotine. Materials and methods: Rat pheochromocytoma 12 (PC12) cells and human neuroglia (HM) cells were used. The ROS levels of the cells were detected by the FACScan. Autophagy flux was monitored by a tandem monomeric RFP-GFP-tagged LC3 lentivirus. The autophagic proteins LC3, SQSTM1/p62 and Beclin1 were detected by western blot assay. In order to evaluate the effects of nicotine and melatonin on the morphological changes of neurons, primary cortical neurons were obtained and immunocytochemistry of TUBB3 tubulin were conducted. Results: Nicotine increased the levels of reactive oxygen species (ROS) in PC12 and HM cells in a concentration-dependent manner. Microscopy showed increased autophagic flux in nicotine-treated PC12 cells. Subsequent western blotting results showed that nicotine induced increase in the levels of LC3B-II and Beclin1, and decreased SQSTM1/p62 in a concentration-dependent manner. Finally, nicotine treatment reduced the length of TUBB3-positive axons and dendrites. Melatonin, a mitochondrially targeted antioxidant, reduced the ROS level, and blocked autophagy activation and the morphologic structural changes induced by nicotine. Conclusions: Our results suggested that the role of nicotine in neuronal toxicity maybe through the induction of ROS and the subsequent activation of autophagy. These effects could be restored by melatonin.