The dysregulation of proliferation and apoptosis plays a significant role in the pathogenesis of postmenopausal osteoporosis (PO). MicroRNAs play an important role in regulating apoptosis of MC3T3-E1 cells. However, the role and potential mechanism of miR-708 for regulating H2O2-induced apoptosis is unknown. This study aimed to investigate the protective function of miR-708 in H2O2-induced apoptosis of MC3T3-E1 osteoblasts. MC3T3-E1 was co-cultured with H2O2 for 8 h, then, flow cytometry, malondialdehyde (MDA), and glutathione peroxidase (Gpx) levels were measured to establish the oxidative model. MiRNA microarray was performed to assess differentially expressed miRNAs between control and H2O2-treated MC3T3-E1 cells. We then performed RT-PCR to identify the relative expression of miR-708 and PTEN. After transfected MC3T3-E1 with miR-708 mimics, flow cytometry, MDA, and Gpx level were performed to identify the apoptosis rate and oxidative stress in these groups. Furthermore, we small interfering RNA of PTEN to identify the role of PTEN in H2O2-induced apoptosis of MC3T3-E1 cells. H2O2 (100 nM) could significantly induce the apoptosis of MC3T3-E1 cells. Moreover, H2O2 could significantly increase the MDA level and downregulated Gpx level. RT-PCR found that H2O2 significantly decrease the level of miR-708. Compared with H2O2 group, H2O2 + miR-708 mimic significantly decreased the apoptosis rate. miR-708 plays a protective role in H2O2-induced MC3T3-E1 osteoblasts apoptosis and its protective effect is proceeded by regulating ROS level and PTEN expression level.

中文翻译：

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MiR-708通过靶向PTEN抑制MC3T3-E1细胞抵抗H2O2诱导的凋亡。

增殖和凋亡的失调在绝经后骨质疏松症（PO）的发病机理中起着重要作用。MicroRNA在调节MC3T3-E1细胞凋亡中起重要作用。但是，miR-708在调节H2O2诱导的细胞凋亡中的作用和潜在机制尚不清楚。本研究旨在探讨miR-708在H2O2诱导的MC3T3-E1成骨细胞凋亡中的保护作用。将MC3T3-E1与H2O2共培养8 h，然后测量流式细胞仪，丙二醛（MDA）和谷胱甘肽过氧化物酶（Gpx）的水平以建立氧化模型。进行MiRNA微阵列分析，以评估对照和H2O2处理的MC3T3-E1细胞之间差异表达的miRNA。然后，我们进行了RT-PCR，以鉴定miR-708和PTEN的相对表达。用miR-708模拟物转染MC3T3-E1后，进行流式细胞仪，MDA和Gpx水平检测，以鉴定这些组的细胞凋亡率和氧化应激。此外，我们小干扰PTEN的RNA，以鉴定PTEN在H2O2诱导的MC3T3-E1细胞凋亡中的作用。H2O2（100 nM）可以明显诱导MC3T3-E1细胞凋亡。此外，H2O2可以显着提高MDA水平和下调Gpx水平。逆转录-聚合酶链反应发现过氧化氢显着降低了miR-708的水平。与H2O2组相比，H2O2 + miR-708模拟物显着降低了细胞凋亡率。miR-708在H2O2诱导的MC3T3-E1成骨细胞凋亡中起保护作用，其保护作用是通过调节ROS水平和PTEN表达水平来实现的。我们用PTEN的小干扰RNA来鉴定PTEN在H2O2诱导的MC3T3-E1细胞凋亡中的作用。H2O2（100 nM）可以明显诱导MC3T3-E1细胞凋亡。此外，H2O2可以显着提高MDA水平和下调Gpx水平。RT-PCR发现过氧化氢显着降低了miR-708的水平。与H2O2组相比，H2O2 + miR-708模拟物显着降低了细胞凋亡率。miR-708在H2O2诱导的MC3T3-E1成骨细胞凋亡中起保护作用，其保护作用是通过调节ROS水平和PTEN表达水平来实现的。我们用PTEN的小干扰RNA来鉴定PTEN在H2O2诱导的MC3T3-E1细胞凋亡中的作用。H2O2（100 nM）可以明显诱导MC3T3-E1细胞凋亡。此外，H2O2可以显着提高MDA水平和下调Gpx水平。RT-PCR发现过氧化氢显着降低了miR-708的水平。与H2O2组相比，H2O2 + miR-708模拟物显着降低了细胞凋亡率。miR-708在H2O2诱导的MC3T3-E1成骨细胞凋亡中起保护作用，其保护作用是通过调节ROS水平和PTEN表达水平来实现的。逆转录-聚合酶链反应发现过氧化氢显着降低了miR-708的水平。与H2O2组相比，H2O2 + miR-708模拟物显着降低了细胞凋亡率。miR-708在H2O2诱导的MC3T3-E1成骨细胞凋亡中起保护作用，其保护作用是通过调节ROS水平和PTEN表达水平来实现的。RT-PCR发现过氧化氢显着降低了miR-708的水平。与H2O2组相比，H2O2 + miR-708模拟物显着降低了细胞凋亡率。miR-708在H2O2诱导的MC3T3-E1成骨细胞凋亡中起保护作用，其保护作用是通过调节ROS水平和PTEN表达水平来实现的。