Roy Abhishek, Sen Santimoy, Das Rudradip, Shard Amit, Kumar Hemant
Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research (NIPER)-Ahmedabad, Gandhinagar, India.
Department of Medicinal Chemistry, National Institute of Pharmaceutical Education and Research (NIPER)- Ahmedabad, Gandhinagar, India.
Neurospine. 2024 Sep;21(3):878-889. doi: 10.14245/ns.2448546.273. Epub 2024 Sep 30.
Spinal cord injury (SCI), one of the major disabilities concerning central nervous system injury, results in permanent tissue loss and neurological impairment. The existing therapeutic options for SCI are limited and predominantly consist of chemical compounds. In this study, we delved into the neuroprotective effects of myricetin, a natural flavonoid compound, and the underlying mechanisms, specifically in the context of SCI, utilizing an in vivo model. Previously, our investigations revealed an elevation in the phosphorylated form of Lin-11, Isl-1, and Mec-3 kinase1 (LIMK1) at chronic time points postinjury, coinciding with neuronal loss and scar formation. Our primary objective here was to assess the potential neuroprotective properties of myricetin in SCI and to ascertain if these effects were linked to LIMK inhibition, a hitherto unexamined pathway to date.
Computational docking and molecular dynamics simulation studies were performed to assess myricetin's potential to bind with LIMK. Then, using a rat contusion model, SCI was induced and different molecular techniques (Western blot, Evans Blue assay, quantitative reverse transcription polymerase chain reaction and immunohistochemistry) were performed to determine the effects of myricetin.
Remarkably, computational docking models identified myricetin as having a better interaction profile with LIMK than standard. Subsequent to myricetin treatment, a significant downregulation in phosphorylated LIMK expression was observed at chronic time points. This reduction correlated with a notable decrease in glial and fibrotic scar formation, and enhanced neuroprotection indicating a positive outcome in vivo.
In summary, our findings underscore myricetin's potential as a bioactive compound capable of attenuating SCI-induced injury cascades by targeting the LIMK pathway.
脊髓损伤（SCI）是中枢神经系统损伤导致的主要残疾之一，会造成永久性组织损失和神经功能障碍。现有的SCI治疗选择有限，主要由化合物组成。在本研究中，我们利用体内模型深入探讨了天然黄酮类化合物杨梅素的神经保护作用及其潜在机制，特别是在SCI背景下。此前，我们的研究发现，在损伤后的慢性时间点，Lin-11、Isl-1和Mec-3激酶1（LIMK1）的磷酸化形式升高，同时伴有神经元损失和瘢痕形成。我们在此的主要目的是评估杨梅素在SCI中的潜在神经保护特性，并确定这些作用是否与LIMK抑制有关，这是迄今为止尚未研究的途径。
进行了计算对接和分子动力学模拟研究，以评估杨梅素与LIMK结合的潜力。然后，使用大鼠挫伤模型诱导SCI，并采用不同的分子技术（蛋白质免疫印迹法、伊文思蓝测定法、定量逆转录聚合酶链反应和免疫组织化学）来确定杨梅素的作用。
值得注意的是，计算对接模型确定杨梅素与LIMK的相互作用比标准物更好。杨梅素治疗后，在慢性时间点观察到磷酸化LIMK表达显著下调。这种减少与胶质和纤维化瘢痕形成的显著减少相关，并增强了神经保护作用，表明在体内有积极的结果。
总之，我们的研究结果强调了杨梅素作为一种生物活性化合物的潜力，它能够通过靶向LIMK途径减轻SCI诱导的损伤级联反应。
