Department of Medical Elementology and Toxicology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, India.
Division of Basic Medical Sciences, Indian Council of Medical Research, Government of India, V. Ramalingaswamy Bhawan, New Delhi 110029, India.
Biochem Pharmacol. 2024 Jun;224:116244. doi: 10.1016/j.bcp.2024.116244. Epub 2024 Apr 27.
Traumatic brain injury (TBI) triggers a bevy of changes including mitochondrial dysfunction, apoptosis, oxidative stress, neurobehavioural impairment, and neuroinflammation, among others. Dantrolene (DNT), a muscle relaxant which inhibits intracellular Ca signaling from the ER, has been repurposed as a potential neuroprotective agent in various neurological diseases. However, there have been limited studies on whether it can mitigate TBI-induced deficits and restore impaired mitochondrial dynamics. This study sought to evaluate whether Dantrolene can potentially provide neuroprotection in an in vivo model of TBI. Male wistar rats subjected to TBI were treated with DNT (10 mg/kg) 1 h and 12 h post surgery. Animals were assessed 24 h post-TBI to evaluate neurobehavioural deficits and cerebral edema. We evaluated the protein expressions of apoptotic, autophagic, and neuroinflammatory markers by immunoblotting, as well as Mitochondrial Membrane Potential (MMP) and Reactive Oxygen Species (ROS) via Flow Cytometry to ascertain the effects of DNT on TBI. We further analysed immunofluorescence staining with Glial Fibrillary Acidic Protein (GFAP) and immunohistochemistry with NF-κβ to investigate neuroinflammation. H&E staining was also performed post-TBI. Our findings revealed DNT administration inhibits mitochondria-mediated apoptotis and reduces heightened oxidative stress. DNT treatment was also found to reverse neurobehavioural impairments and offer neuroprotection by preserving neuronal architechture. We also demonstrated that DNT inhibits neuronal autophagy and alleviates neuroinflammation following TBI by modulating the NF-κβ/Akt signaling pathway. Thus, our results suggest a novel application of DNT in ameliorating the multitude of deficits induced by TBI, thereby conferring neuroprotection.
创伤性脑损伤（TBI）引发多种变化，包括线粒体功能障碍、细胞凋亡、氧化应激、神经行为损伤和神经炎症等。丹曲林（Dantrolene，DNT）是一种肌肉松弛剂，可抑制内质网的细胞内 Ca 信号传导，已被重新用作各种神经疾病的潜在神经保护剂。然而，关于它是否能减轻 TBI 引起的缺陷并恢复受损的线粒体动力学，研究有限。本研究旨在评估丹曲林是否能在 TBI 的体内模型中提供神经保护。雄性 Wistar 大鼠接受 TBI 手术后 1 小时和 12 小时给予 DNT（10mg/kg）治疗。在 TBI 后 24 小时评估动物以评估神经行为缺陷和脑水肿。我们通过免疫印迹评估凋亡、自噬和神经炎症标志物的蛋白表达，通过流式细胞术评估线粒体膜电位（MMP）和活性氧（ROS），以确定 DNT 对 TBI 的影响。我们还进一步分析了胶质纤维酸性蛋白（GFAP）的免疫荧光染色和 NF-κβ的免疫组织化学，以研究神经炎症。TBI 后还进行了 H&E 染色。我们的研究结果表明，DNT 给药抑制线粒体介导的细胞凋亡并降低氧化应激水平。还发现 DNT 治疗可通过保护神经元结构来逆转神经行为损伤并提供神经保护。我们还证明，DNT 通过调节 NF-κβ/Akt 信号通路抑制 TBI 后的神经元自噬并减轻神经炎症。因此，我们的结果表明 DNT 在改善 TBI 引起的多种缺陷方面具有新的应用，从而提供神经保护。
