Department of Biochemistry, Faculty of Medicine, Pusat Perubatan Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Kuala Lumpur 56000, Malaysia.
Clinical Laboratory Section, Institute of Medical Science Technology, Universiti Kuala Lumpur, A1-1, Jalan TKS 1, Taman Kajang Sentral, Kajang 43000, Selangor, Malaysia.
Int J Mol Sci. 2022 Jul 28;23(15):8331. doi: 10.3390/ijms23158331.
Changes in mitochondrial bioenergetics are believed to take place during osteoclastogenesis. This study aims to assess changes in mitochondrial bioenergetics and reactive oxygen species (ROS) levels during polyethylene (PE)-induced osteoclastogenesis in vitro. For this purpose, RAW264.7 cells were cultured for nine days and allowed to differentiate into osteoclasts in the presence of PE and RANKL. The total TRAP-positive cells, resorption activity, expression of osteoclast marker genes, ROS level, mitochondrial bioenergetics, glycolysis, and substrate utilization were measured. The effect of tocotrienols-rich fraction (TRF) treatment (50 ng/mL) on those parameters during PE-induced osteoclastogenesis was also studied. During PE-induced osteoclastogenesis, as depicted by an increase in TRAP-positive cells and gene expression of osteoclast-related markers, higher proton leak, higher extracellular acidification rate (ECAR), as well as higher levels of ROS and NADPH oxidases (NOXs) were observed in the differentiated cells. The oxidation level of some substrates in the differentiated group was higher than in other groups. TRF treatment significantly reduced the number of TRAP-positive osteoclasts, bone resorption activity, and ROS levels, as well as modulating the gene expression of antioxidant-related genes and mitochondrial function. In conclusion, changes in mitochondrial bioenergetics and substrate utilization were observed during PE-induced osteoclastogenesis, while TRF treatment modulated these changes.
线粒体生物能学的变化被认为发生在破骨细胞形成过程中。本研究旨在评估体外聚乙烯(PE)诱导的破骨细胞形成过程中线粒体生物能学和活性氧(ROS)水平的变化。为此,将 RAW264.7 细胞培养 9 天,并在 PE 和 RANKL 的存在下使其分化为破骨细胞。测量总 TRAP 阳性细胞、吸收活性、破骨细胞标记基因的表达、ROS 水平、线粒体生物能学、糖酵解和底物利用。还研究了生育三烯酚丰富馏分(TRF)处理(50ng/mL)对 PE 诱导的破骨细胞形成过程中这些参数的影响。在 PE 诱导的破骨细胞形成过程中,TRAP 阳性细胞和破骨细胞相关标记基因的表达增加,分化细胞中观察到质子泄漏增加、细胞外酸化率(ECAR)升高以及 ROS 和 NADPH 氧化酶(NOXs)水平升高。分化组中一些底物的氧化水平高于其他组。TRF 处理显著减少了 TRAP 阳性破骨细胞的数量、骨吸收活性和 ROS 水平,并调节了抗氧化相关基因和线粒体功能的基因表达。总之,在 PE 诱导的破骨细胞形成过程中观察到线粒体生物能学和底物利用的变化,而 TRF 处理调节了这些变化。