Institut für Pharmakologie und Klinische Pharmazie, Biochemisch-Pharmakologisches Centrum Marburg, Philipps-Universität Marburg, Karl-von-Frisch-Straße 1, Marburg 35032, Germany; Center for Mind Brain and Behavior, Marburg 35032, Germany.
Preclinical Research, Max Zeller Soehne AG, Romanshorn, Switzerland.
Phytomedicine. 2019 Jan;52:107-116. doi: 10.1016/j.phymed.2018.09.177. Epub 2018 Sep 17.
Cimicifuga racemosa extract is a well-established therapy for menopausal symptoms. The mechanisms underlying the multiple therapeutic effects of Cimicifuga extract, e.g. reducing hot flushes and profuse sweating are not well defined. Recent studies revealed pronounced effects of Ze 450, a Cimicifuga racemosa extract that was produced by a standardized procedure, on energy metabolism through activation of AMP-activated protein kinase in vitro and beneficial anti-diabetic effects in vivo.
The aim of the study was to investigate the effects of Ze 450 on energy metabolism. Since mitochondria are the key regulators of cellular energy homeostasis, we wanted to elucidate whether Ze 450 affects mitochondrial resilience and can provide protection against oxidative damage in neuronal and liver cells.
METHODS/STUDY DESIGN: In this study, we investigated the effects of Ze 450 (1-200 µg/ml) on mitochondrial integrity and function, and cell viability in models of oxidative stress induced by erastin and RSL-3 in neuronal and liver cells. The effects of Ze 450 in control conditions and after induction of oxidative stress were analyzed using FACS for detecting lipid peroxidation (BODIPY), mitochondrial ROS formation (MitoSOX), mitochondrial membrane potential (TMRE) and cell death (AnnexinV/PI staining). Furthermore, we determined metabolic activity (MTT assay), ATP levels and mitochondrial respiration and glycolysis (oxygen consumption rates, extracellular acidification rates; Seahorse).
Ze 450 preserved mitochondrial integrity and ATP levels, and prevented mitochondrial ROS formation, loss of mitochondrial membrane potential and cell death. Notably, Cimicifuga racemosa extract alone did not alter mitochondrial ROS levels, and subtle inhibitory effects on cell proliferation were reversed after withdrawal of the extract. In addition, Ze 450 did not exert toxic effects to liver cells, but rather protected these from the oxidative challenge. Further analysis of the mitochondrial oxygen consumption rate and the extracellular acidification rate revealed that Ze 450 mediated a switch from mitochondrial respiration to glycolysis, and this metabolic shift was a prerequisite for the protective effects against oxidative damage.
In conclusion, the bioenergetic shift induced by Ze 450 exerted protective effects in different cell types, and offers promising therapeutic potential in age related diseases involving oxidative stress and mitochondrial damage.
黑升麻提取物是一种被广泛认可的治疗更年期症状的药物。其多种治疗效果的机制，如减少热潮红和大量出汗，尚未得到明确界定。最近的研究表明，由标准化程序生产的黑升麻提取物 Ze 450 对能量代谢具有显著影响，通过体外激活 AMP 激活的蛋白激酶，以及体内的有益的抗糖尿病作用。
本研究旨在探讨 Ze 450 对能量代谢的影响。由于线粒体是细胞能量平衡的关键调节剂，我们想阐明 Ze 450 是否影响线粒体的弹性，并能为神经元和肝细胞提供抵抗氧化损伤的保护。
方法/研究设计：在这项研究中，我们研究了 Ze 450（1-200μg/ml）对神经元和肝细胞中由 erastin 和 RSL-3 诱导的氧化应激模型中线粒体完整性和功能以及细胞活力的影响。使用流式细胞术检测脂质过氧化（BODIPY）、线粒体 ROS 形成（MitoSOX）、线粒体膜电位（TMRE）和细胞死亡（AnnexinV/PI 染色），分析 Ze 450 在对照条件下和诱导氧化应激后的作用。此外，我们还测定了代谢活性（MTT 测定）、ATP 水平以及线粒体呼吸和糖酵解（耗氧量、细胞外酸化率； Seahorse）。
Ze 450 保持了线粒体的完整性和 ATP 水平，并防止了线粒体 ROS 的形成、线粒体膜电位的丧失和细胞死亡。值得注意的是，黑升麻提取物本身并没有改变线粒体 ROS 水平，并且在提取物被撤回后，对细胞增殖的轻微抑制作用得到了逆转。此外，Ze 450 对肝细胞没有毒性作用，反而能保护这些细胞免受氧化应激的影响。进一步分析线粒体耗氧量和细胞外酸化率表明，Ze 450 介导了从线粒体呼吸到糖酵解的代谢转变，这种代谢转变是对氧化损伤产生保护作用的前提。
总之，Ze 450 诱导的生物能量转移在不同的细胞类型中发挥了保护作用，并为涉及氧化应激和线粒体损伤的与年龄相关的疾病提供了有前景的治疗潜力。
