Apelin 通过 TFEB 增强自噬通量促进氧化应激下人退变 NP 细胞的 ECM 合成。
Apelin Promotes ECM Synthesis by Enhancing Autophagy Flux via TFEB in Human Degenerative NP Cells under Oxidative Stress.
机构信息
Department of Orthopedics, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Department of Obstetrics, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
出版信息
Biomed Res Int. 2020 Feb 14;2020:4897170. doi: 10.1155/2020/4897170. eCollection 2020.
BACKGROUND
Apelin alleviates oxidative stress which contributes to the development of aging. IVDD is a disease closely correlated to aging and oxidative stress which is known to be harmful to NP cells' matrix synthesis. The purpose of the present study was to investigate the role and underlying mechanism of Apelin in NP cells' matrix degradation under oxidative stress.
METHODS
First, the mRNA and protein expressions of Apelin were checked by RT-PCR and Western blot in NP from normal and degenerative IVD to explore the relationship between Apelin and IVDD preliminarily. Then, HO was used to mimic oxidative stress of NP cells. After treated with Apelin 13 and CQ, the GAG content was assessed by DMMB and the mRNA/protein expressions of NP matrix macromolecules (Collagen II and Aggrecan) and autophagy-related markers (LC3 and p62) were assessed by RT-PCR/Western blot. Finally, TFEB was knocked down by esiRNA-TFEB transfection and the nucleoprotein expression of TFEB and autophagy-related markers (LC3 and p62) were assessed by Western blot to discuss whether TFEB is involved in Apelin regulating autophagy flux in NP cells under oxidative stress.
RESULTS
Our data first confirmed that the mRNA and protein expressions of Apelin were decreased with IVDD. Furthermore, Apelin increased GAG content of NP cells and mRNA/protein expressions of NP matrix macromolecules (Collagen II and Aggrecan) and promoted autophagic flux (LC3II/I increased and p62 decreased) under oxidative stress. Finally, after transfected with esiRNA-TFEB, Apelin cannot promote autophagic flux any more in human degenerative NP cells.
CONCLUSION
Our data indicated that Apelin promotes ECM synthesis by enhancing autophagy flux via TFEB in human degenerative NP cells under oxidative stress. This viewpoint may provide a new therapeutic idea for IVDD.
背景
Apelin 可减轻氧化应激，这有助于衰老的发展。IVDD 是一种与衰老和氧化应激密切相关的疾病，已知对 NP 细胞基质合成有害。本研究旨在探讨 Apelin 在 NP 细胞氧化应激下基质降解中的作用及其潜在机制。
方法
首先，通过 RT-PCR 和 Western blot 检测正常和退变椎间盘 NP 中的 Apelin mRNA 和蛋白表达，初步探讨 Apelin 与 IVDD 的关系。然后，用 HO 模拟 NP 细胞的氧化应激。用 Apelin 13 和 CQ 处理后，用 DMMB 测定 GAG 含量，用 RT-PCR/Western blot 测定 NP 基质大分子（Collagen II 和 Aggrecan）和自噬相关标志物（LC3 和 p62）的 mRNA/蛋白表达。最后，用 esiRNA-TFEB 转染敲低 TFEB，用 Western blot 检测 TFEB 和自噬相关标志物（LC3 和 p62）的核蛋白表达，探讨 TFEB 是否参与 Apelin 调节 NP 细胞氧化应激下的自噬流。
结果
我们的数据首次证实，Apelin 的 mRNA 和蛋白表达随 IVDD 而降低。此外，Apelin 增加了 NP 细胞的 GAG 含量和 NP 基质大分子（Collagen II 和 Aggrecan）的 mRNA/蛋白表达，并促进了自噬流（LC3II/I 增加，p62 减少）。最后，转染 esiRNA-TFEB 后，Apelin 不能再促进人退变 NP 细胞的自噬流。
结论
我们的数据表明，Apelin 通过 TFEB 增强自噬流促进 ECM 合成，在氧化应激下人退变 NP 细胞。这一观点可能为 IVDD 提供新的治疗思路。
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