Li Zemeng, Ren Dangli, Wang Jingjing, Wang Yatao, Chen Yueyang, Diao Yunfeng, Li Jianwei, Qu Yang, Zheng Maohua, Sun Hongtao
Tianjin Key Laboratory of Neurotrauma Repair, Institute of Traumatic Brain Injury and Neuroscience, Characteristic Medical Center of Chinese People's Armed Police Force, Tianjin, China.
The First Clinical Medical College of Lanzhou University, Lanzhou, China.
PLoS One. 2025 May 30;20(5):e0323259. doi: 10.1371/journal.pone.0323259. eCollection 2025.
As one of the major public health security problems, traumatic brain injury (TBI) is characterized by cerebral dysfunction. The following neuroinflammation is considered as the main secondary injury factor. Targeting the expression of inflammatory cytokines could be effective in alleviating TBI-induced neuroinflammation. The anti-inflammatory role of natural products is increasingly receiving attention. 3-Deoxysappanchalcone (3-DSC) is a bioactive compound from Caesalpinia sappan L.
The present study was designed to investigate the impact of 3-DSC on neuroinflammation in primary microglia and TBI models. To assess cytotoxicity, cell viability tests were conducted with varying concentrations of 3-DSC ranging from 5 to 20 μM. Quantitative PCR (qPCR) and Enzyme-Linked Immunosorbent Assay (ELISA) were utilized to measure the production of inflammatory cytokines in LPS-activated primary microglia treated with or without 3-DSC (at 10 μM). Immune blotting arrays were used to examine the activation of canonical inflammation signaling pathways. To further elucidate the anti-inflammation effect of 3-DSC, RNA-seq was carried out between LPS and LPS + 3-DSC group. In vitro co-culture experiments were carried out to evaluate the protective effect of 3-DSC on neurons against inflammation-mediated apoptosis. Additionally, in vivo experiments were performed to observe the impact of 3-DSC on TBI-induced microglia activation and spatial memory impairment. 3-DSC (160 μg/kg, 320 μg/kg) were administered via the tail vein at day 1 after TBI (n = 6). Behavioral tests were conducted 7 days after traumatic brain injury (TBI) to detect the spatial memory ability of rats.
The cell viability results revealed that within the concentration range of 5-20 μM, 3-DSC did not cause significant cytotoxicity. In the qPCR and ELISA assays, it was found that 3-DSC at 10 μM led to a reduction in the production of inflammatory cytokines. The immune blotting arrays demonstrated that 3-DSC inhibited the activation of NF-kB and MAPK signaling pathways. The results of RNA sequencing revealed the altered signaling pathways and key hub genes. The in vitro co-culture outcomes indicated that 3-DSC could safeguard neurons from apoptosis caused by neuroinflammation. Finally, the in vivo experiments showed that 3-DSC was effective in alleviating TBI-induced microglia activation and spatial memory impairment.
Collectively, these findings suggest that 3-DSC holds promise as a potential compound for the development of therapeutic and preventive agents aimed at treating neuroinflammation-related disorders. It offers a new avenue for further research and potential clinical applications in the context of TBI and neuroinflammation related disorders.
创伤性脑损伤（TBI）作为主要的公共卫生安全问题之一，其特征为脑功能障碍。随后发生的神经炎症被认为是主要的继发性损伤因素。针对炎性细胞因子的表达可能有效减轻TBI诱导的神经炎症。天然产物的抗炎作用日益受到关注。3-去氧苏木查尔酮（3-DSC）是苏木的一种生物活性化合物。
本研究旨在探讨3-DSC对原代小胶质细胞和TBI模型中神经炎症的影响。为评估细胞毒性，使用浓度范围为5至20μM的不同浓度3-DSC进行细胞活力测试。利用定量聚合酶链反应（qPCR）和酶联免疫吸附测定（ELISA）来测量用或不用3-DSC（10μM）处理的脂多糖（LPS）激活的原代小胶质细胞中炎性细胞因子的产生。免疫印迹阵列用于检测经典炎症信号通路的激活。为进一步阐明3-DSC的抗炎作用，在LPS组和LPS + 3-DSC组之间进行了RNA测序。进行体外共培养实验以评估3-DSC对神经元免受炎症介导的凋亡的保护作用。此外，进行体内实验以观察3-DSC对TBI诱导的小胶质细胞激活和空间记忆损伤的影响。TBI后第1天通过尾静脉给予3-DSC（160μg/kg，320μg/kg）（n = 6）。在创伤性脑损伤（TBI）7天后进行行为测试以检测大鼠的空间记忆能力。
细胞活力结果显示，在5-20μM的浓度范围内，3-DSC未引起明显的细胞毒性。在qPCR和ELISA分析中，发现10μM的3-DSC导致炎性细胞因子产生减少。免疫印迹阵列表明3-DSC抑制NF-κB和丝裂原活化蛋白激酶（MAPK）信号通路的激活。RNA测序结果揭示了信号通路和关键枢纽基因的改变。体外共培养结果表明，3-DSC可保护神经元免受神经炎症引起的凋亡。最后，体内实验表明，3-DSC可有效减轻TBI诱导的小胶质细胞激活和空间记忆损伤。
总体而言，这些发现表明，3-DSC有望成为开发治疗和预防神经炎症相关疾病的潜在化合物。它为TBI和神经炎症相关疾病背景下的进一步研究和潜在临床应用提供了新途径。
