白藜芦醇通过激活 AMP 激活的蛋白激酶保护成骨细胞免受地塞米松诱导的细胞毒性。
Resveratrol Protects Osteoblasts Against Dexamethasone-Induced Cytotoxicity Through Activation of AMP-Activated Protein Kinase.
机构信息
Department of Orthopedics, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
Shandong University Spine and Spine Cord Disease Research Center, Shandong University, Jinan, Shandong, People's Republic of China.
出版信息
Drug Des Devel Ther. 2020 Oct 23;14:4451-4463. doi: 10.2147/DDDT.S266502. eCollection 2020.
PURPOSE
Glucocorticoids are used for the treatment of inflammatory diseases, but glucocorticoid treatment is associated with bone damage. Resveratrol is a phytoalexin found in many plants, and we investigated its protective role on dexamethasone-induced dysfunction in MC3T3-E1 cells and primary osteoblasts.
MATERIALS AND METHODS
MC3T3-E1 cells and primary osteoblasts were treated with dexamethasone in the presence/absence of different doses of resveratrol for 24 or 48 h. Then, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium (MTT) and lactate dehydrogenase (LDH) assays were used to evaluate cell viability. Apoptosis was analyzed by a flow cytometry. An alkaline phosphatase (ALP) activity assay and Alizarin Red S staining were used to study osteoblast differentiation. Expression of osteoblast-related genes was measured by real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The AMP-activated protein kinase (AMPK) signaling pathway and mitochondrial expression of superoxide dismutase were evaluated by Western blotting. Intracellular reactive oxygen species (ROS), adenosine triphosphate (ATP) content, mitochondrial-complex activity, and mitochondrial DNA content were measured to evaluate mitochondrial function.
RESULTS
Resveratrol induced the proliferation and inhibited apoptosis of osteoblasts in the presence of dexamethasone. Resveratrol increased the ALP activity and mineralization of osteoblasts. Resveratrol also attenuated dexamethasone-induced inhibition of mRNA expression of osteogenesis maker genes, including bone morphogenetic protein-2, osteoprotegerin, runt-related transcription factor-2, and bone Gla protein. Resveratrol alleviated dexamethasone-induced mitochondrial dysfunction. Resveratrol strongly stimulated expression of peroxisome proliferator-activated receptor-γ coactivator 1α and sirtuin-3 genes, as well as their downstream target gene superoxide dismutase-2. Resveratrol induced phosphorylation of AMPK and acetyl-CoA carboxylase (ACC). Blockade of AMPK signaling using compound C reversed the protective effects of resveratrol against dexamethasone.
CONCLUSION
Resveratrol showed protective effects against dexamethasone-induced dysfunction of osteoblasts by activating AMPK signaling.
目的
糖皮质激素用于治疗炎症性疾病，但糖皮质激素治疗与骨损伤有关。白藜芦醇是一种植物抗毒素，存在于许多植物中，我们研究了它对地塞米松诱导的 MC3T3-E1 细胞和原代成骨细胞功能障碍的保护作用。
材料和方法
用地塞米松处理 MC3T3-E1 细胞和原代成骨细胞，同时存在/不存在不同剂量的白藜芦醇，处理 24 或 48 小时。然后，通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基四氮唑（MTT）和乳酸脱氢酶（LDH）测定评估细胞活力。通过流式细胞术分析细胞凋亡。碱性磷酸酶（ALP）活性测定和茜素红 S 染色用于研究成骨细胞分化。通过实时逆转录定量聚合酶链反应（RT-qPCR）测量成骨细胞相关基因的表达。通过 Western 印迹评估 AMP 激活蛋白激酶（AMPK）信号通路和超氧化物歧化酶的线粒体表达。测量细胞内活性氧（ROS）、三磷酸腺苷（ATP）含量、线粒体复合物活性和线粒体 DNA 含量，以评估线粒体功能。
结果
白藜芦醇在存在地塞米松的情况下诱导成骨细胞增殖并抑制其凋亡。白藜芦醇增加了成骨细胞的 ALP 活性和矿化。白藜芦醇还减轻了地塞米松诱导的成骨基因表达的抑制，包括骨形态发生蛋白-2、骨保护素、 runt 相关转录因子-2 和骨 Gla 蛋白。白藜芦醇减轻了地塞米松诱导的线粒体功能障碍。白藜芦醇强烈刺激过氧化物酶体增殖物激活受体-γ共激活因子 1α 和 Sirtuin-3 基因及其下游靶基因超氧化物歧化酶-2 的表达。白藜芦醇诱导 AMPK 和乙酰辅酶 A 羧化酶（ACC）的磷酸化。使用化合物 C 阻断 AMPK 信号通路会逆转白藜芦醇对地塞米松的保护作用。
结论
白藜芦醇通过激活 AMPK 信号通路对地塞米松诱导的成骨细胞功能障碍表现出保护作用。
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