Song Woo-Ju, Yun Jang-Hyuk, Jeong Myeong-Seon, Kim Kil-Nam, Shin Taekyun, Kim Hyoung-Chun, Wie Myung-Bok
College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon 24341, Korea.
Chuncheon Center, Korea Basic Science Institute, Chuncheon 24341, Korea.
Brain Sci. 2021 Aug 24;11(9):1116. doi: 10.3390/brainsci11091116.
Trimethyltin (TMT) is an environmental neurotoxin that mediates dopaminergic neuronal injury in the brain. In this study, we characterized the toxic mechanism and possible protective compounds against TMT-induced neurotoxicity in human dopaminergic neuroblastoma SH-SY5Y cells. Antioxidants such as melatonin, N-acetylcysteine (NAC), α-tocopherol, and allopurinol alleviated TMT toxicity. Apoptosis induced by TMT was identified by altered expression of cleaved caspase-3, Bax, Bcl-2, and Bcl-xL through Western blot analysis. The iron chelator deferoxamine ameliorated the alteration of apoptosis-related proteins through TMT exposure. TMT also induced delayed ultrastructural necrotic features such as mitochondrial swelling and cytoplasmic membrane rupture; NAC reduced these necrotic injuries. Esculetin, meloxicam, celecoxib, and phenidone decreased TMT toxicity. Elevation of the pro-inflammatory cytokines IL-1β, TNF-α, and NF-ĸB and reduction of the antioxidant enzymes catalase and glutathione peroxidase-1 (GPx-1) were induced by TMT and ameliorated by inhibitors of LOX and COX-2 enzymes. Both NMDA and non-NMDA antagonists attenuated TMT toxicity. The free calcium ion modulators nimodipine and BAPTA/AM contributed to neuronal survival against TMT toxicity. Inhibitors of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway, an autophagy regulator, decreased TMT toxicity. These results imply that TMT neurotoxicity is the chief participant in LOX- and COX-2-mediated apoptosis, partly via necrosis and autophagy in SH-SY5Y cells.
三甲基锡(TMT)是一种环境神经毒素,可介导大脑中多巴胺能神经元损伤。在本研究中,我们对人多巴胺能神经母细胞瘤SH-SY5Y细胞中TMT诱导的神经毒性的作用机制及可能的保护化合物进行了表征。褪黑素、N-乙酰半胱氨酸(NAC)、α-生育酚和别嘌呤醇等抗氧化剂可减轻TMT毒性。通过蛋白质免疫印迹分析检测裂解的半胱天冬酶-3、Bax、Bcl-2和Bcl-xL表达的改变,从而确定TMT诱导的细胞凋亡。铁螯合剂去铁胺可改善TMT暴露引起的凋亡相关蛋白的改变。TMT还诱导了延迟的超微结构坏死特征,如线粒体肿胀和细胞质膜破裂;NAC减少了这些坏死损伤。七叶亭、美洛昔康、塞来昔布和非那吡啶可降低TMT毒性。TMT可诱导促炎细胞因子IL-1β、TNF-α和NF-κB升高,以及抗氧化酶过氧化氢酶和谷胱甘肽过氧化物酶-1(GPx-1)降低,而脂氧合酶(LOX)和环氧化酶-2(COX-2)抑制剂可改善这种情况。NMDA和非NMDA拮抗剂均可减弱TMT毒性。游离钙离子调节剂尼莫地平和BAPTA/AM有助于神经元抵抗TMT毒性存活。自噬调节剂磷酸肌醇3-激酶/蛋白激酶B/雷帕霉素哺乳动物靶标通路抑制剂可降低TMT毒性。这些结果表明,TMT神经毒性是LOX和COX-2介导的细胞凋亡的主要参与者,部分是通过SH-SY5Y细胞中的坏死和自噬实现的。