Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Beijing Institute of Radiation Medicine, Beijing, 100850, China.
Beijing Institute of Radiation Medicine, Beijing, 100850, China.
J Ethnopharmacol. 2021 Apr 24;270:113765. doi: 10.1016/j.jep.2020.113765. Epub 2021 Jan 5.
Aconite is a processed product of seminal root of perennial herbaceous plant Aconitum Carmichaclii Debx. of Ranunculaceae. It has the effects of warming and tonifying heart yang and restoring yang to save from collapse. Aconitine is the main effective constituent of aconite and used to prevent and treat heart disease. However, how aconitine exerts myocardial protection is still poorly understood.
The present study aimed to investigate the effects of aconitine on mitochondrial dysfunction and explore its mechanism of action.
The model of myocardial injury was induced by Angiotensin II (Ang II) (1 × 10 mol L), and H9c2 cells were incubated with different concentrations of aconitine. The effect of aconitine on mitochondrial was determined by flow cytometry, transmission electron microscopy, luciferase, Seahorse technique and Western blot. The effects of aconitine on sirtuin-3 (Sirt3) activity and Cyclophilin D (CypD) acetylation were detected by immunofluorescence, RT-PCR and co-immunoprecipitation.
We demonstrate that aconitine alleviates the energy metabolic dysfunction of H9c2 cells by activating Sirt3 to deacetylate CypD and inhibiting mitochondrial permeability transition pore (mPTP) opening. In cardiomyocytes, aconitine significantly reduced mitochondrial fragmentation, inhibited acetylation of CypD, suppressed the mPTP opening, mitigated mitochondrial OXPHOS disorders, and improved the synthesis ability of ATP. In contrast, Sirt3 deficiency abolished the effects of aconitine on mPTP and OXPHOS, indicating that aconitine improves mitochondrial function by activating Sirt3.
These results showed that aconitine attenuated the energy metabolism disorder by promoting Sirt3 expression and reducing CypD-mediated mPTP excess openness, rescuing mitochondrial function. Improve mitochondrial function may be a therapeutic approach for treating heart disease, which will generate fresh insight into the cardioprotective of aconitine.
乌头是毛茛科多年生草本植物乌头的块根经炮制而成的制品。它具有温阳强心、回阳救逆的功效。乌头碱是乌头的主要有效成分，用于预防和治疗心脏病。然而，乌头碱如何发挥心肌保护作用还知之甚少。
本研究旨在探讨乌头碱对线粒体功能障碍的影响，并探讨其作用机制。
用血管紧张素 II（Ang II）（1×10-5mol/L）诱导心肌损伤模型，用不同浓度的乌头碱孵育 H9c2 细胞。用流式细胞术、透射电镜、荧光素酶、 Seahorse 技术和 Western blot 检测乌头碱对线粒体的影响。用免疫荧光、RT-PCR 和免疫共沉淀检测乌头碱对 Sirtuin-3（Sirt3）活性和环孢菌素 D（CypD）乙酰化的影响。
我们证明乌头碱通过激活 Sirt3 使 CypD 去乙酰化并抑制线粒体通透性转换孔（mPTP）开放，从而缓解 H9c2 细胞的能量代谢功能障碍。在心肌细胞中，乌头碱显著减少线粒体片段化，抑制 CypD 乙酰化，抑制 mPTP 开放，减轻线粒体 OXPHOS 障碍，提高 ATP 的合成能力。相反，Sirt3 缺陷消除了乌头碱对 mPTP 和 OXPHOS 的作用，表明乌头碱通过激活 Sirt3 改善线粒体功能。
这些结果表明，乌头碱通过促进 Sirt3 表达和减少 CypD 介导的 mPTP 过度开放来减轻能量代谢障碍，从而挽救线粒体功能。改善线粒体功能可能是治疗心脏病的一种治疗方法，这将为乌头碱的心肌保护作用提供新的见解。
