Acute dose of melatonin via Nrf2 dependently prevents acute ethanol-induced neurotoxicity in the developing rodent brain.
Acute dose of melatonin via Nrf2 dependently prevents acute ethanol-induced neurotoxicity in the developing rodent brain.
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DOI：
10.1186/s12974-018-1157-x
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发表时间：
2018-04-21
影响因子：
9.3
通讯作者：
Kim MO
中科院分区：
文献类型：
--
作者：
Ali T;Rehman SU;Shah FA;Kim MO
褪黑激素是一种强有力的内源性抗氧化剂，具有显著的神经保护作用。在目前的研究中，我们探讨了核因子红细胞2相关因子2（Nrf 2）基因依赖的抗氧化机制的神经保护作用的急性褪黑激素对急性乙醇诱导的活性氧（ROS）介导的神经炎症和神经退行性病变在发育中的啮齿动物大脑。将体内大鼠幼仔用单剂量的急性乙醇（5 g/kg，皮下（S.C.））和单剂量的急性褪黑激素（20 mg/kg，腹膜内（I. P.））。一次皮下注射后四小时。和腹膜内注射后，处死所有大鼠幼仔用于进一步的生物化学（Western印迹、ROS测定、LPO测定和免疫组织化学）分析。为了证实体内结果，我们使用了体外小鼠海马HT 22和小胶质细胞BV 2细胞，这些细胞用Nrf 2基因的小干扰RNA（siRNA）敲低，并暴露于褪黑激素（100 μM）和乙醇（100 mM），然后进行进一步的生化分析。我们的生化，免疫组化和免疫荧光结果表明，急性褪黑激素显着上调主内源性抗氧化剂Nrf 2和血红素加氧酶-1，从而扭转急性乙醇诱导的ROS和氧化应激升高的发展中啮齿动物的大脑，并在小鼠海马HT 22和小胶质细胞BV 2细胞。此外，急性褪黑激素随后通过降低活化的胶质细胞增生的表达减少活化的MAPK-p-P38-JNK通路并减轻神经炎症，下调p-NF-K-B/p-IKKβ通路并降低发育中啮齿动物脑和BV 2细胞中其他炎症标志物的表达水平。值得注意的是，褪黑激素通过Nrf 2依赖性机制来减弱出生后啮齿动物脑和HT 22细胞中的神经元凋亡。免疫组织荧光结果还表明，褪黑激素防止乙醇诱导的神经退行性病变在发育中的啮齿动物的大脑。体外实验结果表明，褪黑素通过Nrf 2依赖的方式诱导神经保护作用，并减轻乙醇诱导的神经毒性。褪黑激素的多效性和有效的神经保护性抗氧化特性，以及我们在体内和体外的研究结果，假设急性褪黑激素可能有利于预防和对抗急性乙醇诱导的神经毒性作用，如ROS升高，神经炎症和发育中的啮齿动物大脑中的神经变性。
Melatonin is a well-known potent endogenous antioxidant pharmacological agent with significant neuroprotective actions. Here in the current study, we explored the nuclear factor erythroid 2-related factor 2 (Nrf2) gene-dependent antioxidant mechanism underlying the neuroprotective effects of the acute melatonin against acute ethanol-induced elevated reactive oxygen species (ROS)-mediated neuroinflammation and neurodegeneration in the developing rodent brain. In vivo rat pups were co-treated with a single dose of acute ethanol (5 g/kg, subcutaneous (S.C.)) and a single dose of acute melatonin (20 mg/kg, intraperitoneal (I.P.)). Four hours after a single S.C. and I.P. injections, all of the rat pups were sacrificed for further biochemical (Western blotting, ROS- assay, LPO-assay, and immunohistochemical) analyses. In order to corroborate the in vivo results, we used the in vitro murine-hippocampal HT22 and microglial BV2 cells, which were subjected to knockdown with small interfering RNA (siRNA) of Nrf2 genes and exposed with melatonin (100 μM) and ethanol (100 mM) and proceed for further biochemical analyses. Our biochemical, immunohistochemical, and immunofluorescence results demonstrate that acute melatonin significantly upregulated the master endogenous antioxidant Nrf2 and heme oxygenase-1, consequently reversing the acute ethanol-induced elevated ROS and oxidative stress in the developing rodent brain, and in the murine-hippocampal HT22 and microglial BV2 cells. In addition, acute melatonin subsequently reduced the activated MAPK-p-P38-JNK pathways and attenuated neuroinflammation by decreasing the expression of activated gliosis and downregulated the p-NF-K-B/p-IKKβ pathway and decreased the expression levels of other inflammatory markers in the developing rodent brain and BV2 cells. Of note, melatonin acted through the Nrf2-dependent mechanism to attenuate neuronal apoptosis in the postnatal rodent brain and HT22 cells. Immunohistofluorescence results also showed that melatonin prevented ethanol-induced neurodegeneration in the developing rodent brain. The in vitro results indicated that melatonin induced neuroprotection via Nrf2-dependent manner and reduced ethanol-induced neurotoxicity. The pleiotropic and potent neuroprotective antioxidant characteristics of melatonin, together with our in vivo and in vitro findings, suppose that acute melatonin could be beneficial to prevent and combat the acute ethanol-induced neurotoxic effects, such as elevated ROS, neuroinflammation, and neurodegeneration in the developing rodent brain.
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DOI：
10.2174/1871527315666151110130139
发表时间：
2016-01-01
影响因子：
3
作者：
Ahmad, Ashfaq;Shah, Shahid A.;Kim, Myeong O.
通讯作者：
Kim, Myeong O.
影响因子：
5.3
作者：
Baydas, G;Tuzcu, M
通讯作者：
Tuzcu, M
影响因子：
2.4
作者：
Gitto, E;Romeo, C;Zuccarello, B
通讯作者：
Zuccarello, B
影响因子：
10.3
作者：
Fischer, Tobias W.;Kleszczynski, Konrad;Zillikens, Detlef
通讯作者：
Zillikens, Detlef
影响因子：
2.6
作者：
Guizzetti M;Zhang X;Goeke C;Gavin DP
通讯作者：
Gavin DP