Department of Endocrinology, The Second Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, China.
The Operative Surgery Laboratory, Bengbu Medical College, Bengbu, Anhui, China.
J Biochem Mol Toxicol. 2021 Jul;35(7):e22778. doi: 10.1002/jbt.22778. Epub 2021 Mar 23.
Coenzyme Q10 (CoQ10) has been reported to improve bone density and the number of trabeculae in postmenopausal osteoporosis, but the mechanism remains to be elucidated. We aimed to investigate the effects of CoQ10 on receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and the underlying molecular mechanisms. RAW264.7 cells were treated with different concentrations of RANKL to differentiate into osteoclasts, and then these cells were treated with different concentrations of CoQ10 with or without H O . Tartrate-resistant acid phosphatase staining was performed to detect osteoclasts. Cell viability was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell apoptosis was examined by flow cytometry, and the effects of CoQ10 on protein and messenger RNA expression of mitochondrial apoptosis-associated proteins and osteoclast marker proteins were measured by quantitative reverse transcription polymerase chain reaction and western blot, respectively. Furthermore, enzyme-linked immunosorbent assay was conducted to analyze the activities of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT). RANKL significantly induced osteoclastogenesis in RAW264.7 cells, with the greatest efficiency at 50 ng/ml. CoQ10 had no significant effects on cell viability but it significantly increased the percentages of cell apoptosis. Mechanically, CoQ10 statistically decreased the levels of Bcl-2 and cytochrome C in mitochondria and upregulated the levels of Bax, cleaved caspase 3, and cytochrome C in the cytoplasm. Moreover, CoQ10 significantly decreased RANKL-induced osteoclastogenesis regardless of exposure to H O . In addition, CoQ10 statistically reduced MDA activity and elevated the activities of SOD and CAT, as well as the expression of oxidative stress-related proteins. CoQ10 may inhibit RANKL-induced osteoclastogenesis by regulation of mitochondrial apoptosis and oxidative stress in RAW264.7 cells.
辅酶 Q10（CoQ10）已被报道可改善绝经后骨质疏松症的骨密度和小梁数量，但作用机制尚不清楚。我们旨在研究 CoQ10 对核因子-κB 受体激活配体（RANKL）诱导的破骨细胞形成的影响及其潜在的分子机制。用不同浓度的 RANKL 处理 RAW264.7 细胞以分化为破骨细胞，然后用不同浓度的 CoQ10 处理这些细胞，有或无 H2O2。用抗酒石酸酸性磷酸酶染色检测破骨细胞。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测细胞活力，通过流式细胞术检测细胞凋亡，通过定量逆转录聚合酶链反应和 Western blot 分别检测 CoQ10 对线粒体凋亡相关蛋白和破骨细胞标志物蛋白的蛋白质和信使 RNA 表达的影响。此外，通过酶联免疫吸附试验分析丙二醛（MDA）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。RANKL 显著诱导 RAW264.7 细胞中的破骨细胞形成，在 50ng/ml 时效率最高。CoQ10 对细胞活力没有显著影响，但显著增加了细胞凋亡的百分比。机制上，CoQ10 统计学上降低了线粒体中 Bcl-2 和细胞色素 C 的水平，并上调了细胞质中 Bax、裂解的 caspase 3 和细胞色素 C 的水平。此外，CoQ10 显著降低了 RANKL 诱导的破骨细胞形成，无论是否暴露于 H2O2。此外，CoQ10 统计学上降低了 MDA 活性，提高了 SOD 和 CAT 的活性以及氧化应激相关蛋白的表达。CoQ10 可能通过调节 RAW264.7 细胞中线粒体凋亡和氧化应激来抑制 RANKL 诱导的破骨细胞形成。
