Department of Orthopedics and Traumatology, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China.
Research Student Academy, Guangxi University of Traditional Chinese Medicine, Nanning, China.
Bioengineered. 2022 Apr;13(4):8657-8666. doi: 10.1080/21655979.2022.2054499.
Hyperoside (Hyp) is a flavonoid active compound deriving from Chinese herbal medicines. Increasing studies have implicated that Hyp may serve as a predominant promoting factor in osteoblast differentiation. This paper investigates whether Hyp could relieve glucocorticoid-induced osteonecrosis of the femoral head (GONFH) via promoting osteoblast survival and differentiation as well as to uncover its potential mechanism. GONFH cell model was induced by treating MC3T3-E1 cells with dexamethasone (DEX). The viability, apoptosis, and osteogenic differentiation of DEX-induced cells with the presence or absence of Hyp were assessed by CCK-8, Tunel, ALP assay, and ARS staining, respectively. The NADPH Oxidase 4 (NOX4) overexpression was performed by transfection with overexpression vector. Besides, western blot was used to determine the levels of apoptosis-, osteogenic differentiation-, and c-Jun N-terminal kinase (JNK) signaling-related proteins. It was noticed that Hyp caused no significant effects on the viability of MC3T3-E1 cells without any treatment but significantly enhanced the viability of DEX-induced cells. Besides, Hyp inhibited the apoptosis in DEX-induced cells but enhanced ALP activity and calcium nodule formation. Additionally, Hyp declined NOX4 expression in DEX-induced cells. However, NOX4 overexpression partially reversed the impacts of Hyp on DEX-exposed MC3T3-E1 cells. Finally, Hyp suppressed the activation of ROS/JNK pathway in DEX-induced cells, which was then counteracted by NOX4 overexpression. In conclusion, Hyp could promote the survival and differentiation of DEX-induced osteoblasts by targeting NOX4 to inhibit the ROS/JNK pathway. These results provide evidence for the application of Hyp in treating GONFH.
高圣草素（Hyp）是一种源自中草药的黄酮类活性化合物。越来越多的研究表明，Hyp 可能是成骨细胞分化的主要促进因素。本文研究 Hyp 是否通过促进成骨细胞存活和分化来缓解糖皮质激素诱导的股骨头坏死（GONFH），并揭示其潜在机制。通过用地塞米松（DEX）处理 MC3T3-E1 细胞来诱导 GONFH 细胞模型。通过 CCK-8、Tunel、ALP 测定和 ARS 染色分别评估 DEX 诱导的细胞在存在或不存在 Hyp 的情况下的活力、凋亡和成骨分化。通过转染过表达载体来进行 NADPH 氧化酶 4（NOX4）过表达。此外，通过 Western blot 测定凋亡、成骨分化和 c-Jun N 末端激酶（JNK）信号相关蛋白的水平。结果表明，Hyp 对未经任何处理的 MC3T3-E1 细胞的活力没有显著影响，但显著增强了 DEX 诱导的细胞的活力。此外，Hyp 抑制了 DEX 诱导的细胞凋亡，但增强了 ALP 活性和钙结节形成。此外，Hyp 降低了 DEX 诱导的细胞中的 NOX4 表达。然而，NOX4 过表达部分逆转了 Hyp 对 DEX 暴露的 MC3T3-E1 细胞的影响。最后，Hyp 抑制了 DEX 诱导的细胞中 ROS/JNK 通路的激活，而过表达 NOX4 则拮抗了这一作用。总之，Hyp 通过靶向 NOX4 抑制 ROS/JNK 通路，促进 DEX 诱导的成骨细胞的存活和分化。这些结果为 Hyp 在治疗 GONFH 中的应用提供了证据。
