阿伦膦酸盐促进由 SP-1/SOX-9 介导的细胞外基质基因表达。

Alendronate promotes the gene expression of extracellular matrix mediated by SP-1/SOX-9.

机构信息

Department of Surgery, 481863Shandong Medical College, Linyi, Shandong, China.

Department of Trauma Surgery, 529858Linyi People's Hospital, Linyi, Shandong, China.

出版信息

Hum Exp Toxicol. 2021 Jul;40(7):1173-1182. doi: 10.1177/0960327120988875. Epub 2021 Feb 1.

DOI:10.1177/0960327120988875

PMID:33522294

Abstract

BACKGROUND AND PURPOSE

Osteoarthritis (OA) is a disease with significant degenerative changes of articular cartilage, which is reported to be closely related to the integrity of chondrocytes extracellular matrix (ECM). Alendronate belongs to the family of bisphosphonates with promising cartilage repair function. In the present study, the effects of Alendronate on the gene expression of chondrocytes ECM and the potential mechanism will be investigated to explore the potential therapeutic property of Alendronate on OA.

METHODS

Human SW1353 chondrocytes were stimulated with 1 and 2 μM Alendronate for 12 h. The gene expression of , and in the treated chondrocytes was determined by qRT-PCR. QRT-PCR and western blot analysis were used to evaluate the expression level of SOX-9 in the treated chondrocytes. The expression level of SP-1 was checked by qRT-PCR and immunostaining. SiRNA against SP-1 was transfected into chondrocytes to knockdown the expression of SP-1. The levels of p-ERK1/2 and total ERK1/2 were examined using western blot analysis. TNF-α was used to induce an OA-like model in the chondrocytes for therapeutic evaluations.

RESULTS

Treatment with Alendronate increased the levels of ECM related genes (, and ) in a dose-dependent manner through increasing the expression of SOX-9, a central regulator of ECM genes. Additionally, our findings demonstrate that the effects of Alendronate in the expression of SOX-9 are mediated by SP-1 as silencing of SP-1 abolished these effects. Notably, Alendronate increased the phosphorylation of ERK1/2 and inhibition of ERK1/2 using its specific inhibitor U0126 blocked the expression of SP-1. Finally, we found that treatment with Alendronate could rescue TNF-α-induced reduction of , and SOX-9.

CONCLUSION

Our data indicated that Alendronate might promote the gene expression of extracellular matrix through SOX-9 mediated by the ERK1/2/SP1 signaling pathway.

摘要

背景与目的

骨关节炎（OA）是一种关节软骨发生显著退行性改变的疾病，据报道与软骨细胞细胞外基质（ECM）的完整性密切相关。阿仑膦酸钠属于双膦酸盐家族，具有有前景的软骨修复功能。在本研究中，我们将研究阿仑膦酸钠对软骨细胞 ECM 基因表达的影响及其潜在机制，以探索阿仑膦酸钠治疗 OA 的潜在治疗特性。

方法

用 1 和 2 μM 阿仑膦酸钠刺激人 SW1353 软骨细胞 12 小时。用 qRT-PCR 测定处理后软骨细胞中、和的基因表达。用 qRT-PCR 和 Western blot 分析评估处理后软骨细胞中 SOX-9 的表达水平。用 qRT-PCR 和免疫染色检查 SP-1 的表达水平。用 SP-1 siRNA 转染软骨细胞以敲低 SP-1 的表达。用 Western blot 分析检测 p-ERK1/2 和总 ERK1/2 的水平。用 TNF-α诱导软骨细胞 OA 样模型进行治疗评估。

结果

阿仑膦酸钠呈剂量依赖性增加 ECM 相关基因（、和）的水平，通过增加 ECM 基因的中央调节因子 SOX-9 的表达。此外，我们的研究结果表明，阿仑膦酸钠在 SOX-9 表达中的作用是由 SP-1 介导的，因为沉默 SP-1 可消除这些作用。值得注意的是，阿仑膦酸钠增加 ERK1/2 的磷酸化，用其特异性抑制剂 U0126 抑制 ERK1/2 可阻断 SP-1 的表达。最后，我们发现阿仑膦酸钠治疗可挽救 TNF-α诱导的、和 SOX-9 表达减少。