Zhu Lingling, Lin Lin, Yang Zhenyu, Tang Hongxia, Wu Jinping, Kang Zeyi, Feng Yi, Zheng Binbin, Hu Qirou, Wang Shanshan, Liu Cuimin, Jiang Zhengli, Liu Suzhi, Wu Gang
Department of Pharmacy, Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University, No.150, Ximen Street, Linhai, 317000, Zhejiang Province, China.
School of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Neurochem Res. 2025 Sep 18;50(5):302. doi: 10.1007/s11064-025-04557-x.
Disease-modifying treatments for Parkinson's disease (PD) are urgently needed, with the Nrf2/ARE pathway a promising target. This study aims to explore the effects of sinomenine on PD and Nrf2/ARE activation.
6-OHDA-treated Parkinsonian SH-SY5Y cells and rats were used. Apoptosis and cell viability were measured using flow cytometry and CCK-8 assays. Nrf2 and its downstream proteins were assessed by Western blotting, while ROS levels were detected with fluorescent dyes. Nrf2 silencing via shRNA evaluated sinomenine's dependence on Nrf2 activation. In vivo, behavioral changes, tyrosine hydroxylase (TH) levels and malondialdehyde (MDA) levels were measured. Microglial inflammation was analyzed by measuring TNF-α and IL-1β expression and cytoskeleton analysis. Nrf2 nuclear translocation was verified by Western blotting and molecular docking was performed.
Sinomenine reduced apoptosis and ROS, improved cell viability, upregulated Nrf2 and antioxidant enzyme expression. The protective effects against apoptosis were abolished by Nrf2 silencing. In PD animals, sinomenine improved motor deficits, enhanced Nrf2, GCLC, GCLM, NQO1, and HO-1 expression, decreased MDA levels, increased TH levels in the striatum and maintained count of dopaminergic neurons in substantia nigra. Additionally, it suppressed TNF-α and IL-1β levels in brain tissue and blood, preserving normal microglial morphology and reducing neuroinflammation. Sinomenine promoted Nrf2 nuclear translocation and showed high Keap1 affinity in docking.
Sinomenine activates the Nrf2/ARE pathway, mitigating oxidative stress and inflammation in PD models, possibly through Keap1 binding and Nrf2 nuclear translocation. These findings suggest sinomenine may serve as a potential disease-modifying therapy for Parkinson's disease, pending further clinical validation.
帕金森病(PD)急需改善病情的治疗方法,核因子E2相关因子2/抗氧化反应元件(Nrf2/ARE)通路是一个有前景的靶点。本研究旨在探讨青藤碱对帕金森病及Nrf2/ARE激活的影响。
使用6-羟基多巴胺(6-OHDA)处理的帕金森病SH-SY5Y细胞和大鼠。采用流式细胞术和CCK-8法检测细胞凋亡和细胞活力。通过蛋白质免疫印迹法评估Nrf2及其下游蛋白,用荧光染料检测活性氧(ROS)水平。通过短发夹RNA(shRNA)沉默Nrf2来评估青藤碱对Nrf2激活的依赖性。在体内,测量行为变化、酪氨酸羟化酶(TH)水平和丙二醛(MDA)水平。通过测量肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)表达以及细胞骨架分析来分析小胶质细胞炎症。通过蛋白质免疫印迹法验证Nrf2核转位并进行分子对接。
青藤碱减少细胞凋亡和ROS,提高细胞活力,上调Nrf2和抗氧化酶表达。Nrf2沉默消除了其对细胞凋亡的保护作用。在帕金森病动物中,青藤碱改善运动功能障碍,增强Nrf2、谷氨酸半胱氨酸连接酶催化亚基(GCLC)、谷氨酸半胱氨酸连接酶调节亚基(GCLM)、醌氧化还原酶1(NQO1)和血红素加氧酶-1(HO-1)表达,降低MDA水平,增加纹状体中TH水平并维持黑质中多巴胺能神经元数量。此外,它抑制脑组织和血液中TNF-α和IL-1β水平,保持正常小胶质细胞形态并减轻神经炎症。青藤碱促进Nrf2核转位并在对接中显示出对Keap1的高亲和力。
青藤碱激活Nrf2/ARE通路,减轻帕金森病模型中的氧化应激和炎症,可能是通过与Keap1结合和Nrf2核转位实现的。这些发现表明,在进一步临床验证之前,青藤碱可能作为帕金森病潜在的改善病情疗法。