Rehabilitation Department, Zhongshan Hospital, Xiamen University, Xiamen, 361004, Fujian, China.
Xiamen Humanity Rehabilitation Hospital, No. 3775 Xianyue Rd, Xiamen, 361006, China.
Calcif Tissue Int. 2024 Nov;115(5):673-685. doi: 10.1007/s00223-024-01271-8. Epub 2024 Aug 28.
To investigate the potential mechanism of Morinda officinalis F. C. How polysaccharides (MOPs) in regulating osteoclast differentiation and apoptosis through miR-214-3p and its target protein. Ovariectomy was performed in 8-week female C57BL6 mice to establish the postmenopausal osteoporosis (PMOP) model. Mice were treated immediately with 500 mg/kg of MOPs (prevention group); others were treated 2 weeks after operation (treatment group). Left femur bone mineral density (BMD) was examined. RAW264.7 cells were administered with receptor activator of NF-κB ligand (RANKL) to establish the osteoclast (OC) model and treated with serum containing 1 or 2 g/kg of MOPs. Apoptosis-related indexes, miR-214-3p, and Expressed Developmentally Down-regulated 4-Like (NEDD4L) were detected by western blot, quantitative real-time-reverse transcription polymerase chain reaction (qRT-PCR), and flow cytometry. OC received a miR-214-3p inhibitor or NEDD4L small interfering RNA (siRNA). MOPs reversed the PMOP-induced changes in bones. Compared with the RANKL group, MOPs increased the apoptosis and related markers in OCs. MOPs decreased the femur miR-214-3p of PMOP mice (P < 0.001). Higher concentrations of MOPs reversed the upregulation of miR-214 mRNA in OCs (P < 0.001). miR-214-3p inhibitor increased the expression of Bax and CC3 (P < 0.01) and decreased the expression of Bcl-2 (P < 0.05). NEDD4L is targeted by miR-214. NEDD4L was upregulated in the RANKL + MOPs group (P < 0.01). miR-214-3p inhibitor increased the upregulation of NEDD4L induced by MOPs (P < 0.05). siRNA NEDD4L significantly reversed the inhibition of MOPs on osteoclast differentiation with miR-214-3p inhibitor (P < 0.01). MOPs effectively prevent PMOP by inhibiting osteoclastogenesis and inducing OC apoptosis through the miR-214-3p/NEDD4L pathway.
为了探究桑枝多糖(MOPs)通过 miR-214-3p 及其靶蛋白调控破骨细胞分化和凋亡的潜在机制。对 8 周龄雌性 C57BL6 小鼠进行卵巢切除术以建立绝经后骨质疏松症(PMOP)模型。术后立即给予 500mg/kg MOPs(预防组);其他组在手术后 2 周给予治疗。检测左股骨骨密度(BMD)。用核因子-κB 受体激活剂配体(RANKL)处理 RAW264.7 细胞建立破骨细胞(OC)模型,并以含 1 或 2g/kg MOPs 的血清处理。通过 Western blot、定量实时逆转录聚合酶链反应(qRT-PCR)和流式细胞术检测凋亡相关指标、miR-214-3p 和表达发育下调 4 样(NEDD4L)。OC 接受 miR-214-3p 抑制剂或 NEDD4L 小干扰 RNA(siRNA)。MOPs 逆转了 PMOP 引起的骨骼变化。与 RANKL 组相比,MOPs 增加了 OCs 的凋亡及相关标志物。MOPs 降低了 PMOP 小鼠股骨 miR-214-3p(P<0.001)。较高浓度的 MOPs 逆转了 OCs 中 miR-214mRNA 的上调(P<0.001)。miR-214-3p 抑制剂增加了 Bax 和 CC3 的表达(P<0.01),降低了 Bcl-2 的表达(P<0.05)。NEDD4L 是 miR-214 的靶标。在 RANKL+MOPs 组中上调 NEDD4L(P<0.01)。miR-214-3p 抑制剂增加了 MOPs 诱导的 NEDD4L 上调(P<0.05)。siRNA NEDD4L 显著逆转了 miR-214-3p 抑制剂对 MOPs 诱导的破骨细胞分化的抑制作用(P<0.01)。MOPs 通过 miR-214-3p/NEDD4L 通路有效抑制破骨细胞生成,诱导 OC 凋亡,从而有效预防 PMOP。