IRE1α inhibits osteogenic differentiation of mouse embryonic fibroblasts by limiting Shh signaling.

IF 2.9 3区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Oral diseases Pub Date : 2024-10-01 Epub Date: 2024-03-04 DOI:10.1111/odi.14919

Zhixiang Zhang, Xuan Zhang, Xiangzhen Wei, Chengbo Yu, Li Xiao, Jianmiao Liu, Yong Liu, Yingguang Cao, Ke Song

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引用次数: 0

Abstract

Objectives: This study aimed to investigate the effect of endoplasmic reticulum (ER) stress sensor inositol-requiring enzyme 1α (IRE1α) on the sonic hedgehog N-terminus (N-Shh)-enhanced-osteogenic differentiation process in mouse embryonic fibroblasts (MEFs).

Materials and methods: Osteogenesis of MEFs was observed by alkaline phosphatase (ALP) staining, alizarin red staining, and Von Kossa staining assays. Activation of unfolded protein response and Shh signaling were examined using real-time quantitative PCR and western blot assays. IRE1α-deficient MEFs were used to explore the effect of IRE1α on N-Shh-driven osteogenesis.

Results: N-Shh increased ALP activity, matrix mineralization, and the expression of Alp and Col-I in MEFs under osteogenic conditions; notably, this was reversed when combined with the ER stress activator Tm treatment. Interestingly, the administration of N-Shh decreased the expression of IRE1α. Abrogation of IRE1α increased the expression of Shh pathway factors in osteogenesis-induced MEFs, contributing to the osteogenic effect of N-Shh. Moreover, IRE1α-deficient MEFs exhibited elevated levels of osteogenic markers.

Conclusions: Our findings suggest that the IRE1α-mediated unfolded protein response may alleviate the ossification of MEFs by attenuating Shh signaling. Our research has identified a strategy to inhibit excessive ossification, which may have clinical significance in preventing temporomandibular joint bony ankylosis.

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IRE1α 通过限制 Shh 信号抑制小鼠胚胎成纤维细胞的成骨分化。

研究目的本研究旨在探讨内质网（ER）应激传感器肌醇需要酶1α（IRE1α）对小鼠胚胎成纤维细胞（MEFs）中声刺猬N-端（N-Shh）增强-成骨分化过程的影响：通过碱性磷酸酶（ALP）染色法、茜素红染色法和冯-科萨染色法观察 MEFs 的成骨过程。通过实时定量 PCR 和 Western 印迹检测未折叠蛋白反应和 Shh 信号的激活情况。利用IRE1α缺陷的MEFs探讨IRE1α对N-Shh驱动的成骨作用：结果：在成骨条件下，N-Shh增加了MEFs中的ALP活性、基质矿化以及Alp和Col-I的表达；值得注意的是，当与ER应激激活剂Tm联合处理时，这种影响被逆转。有趣的是，给予 N-Shh 会降低 IRE1α 的表达。抑制 IRE1α 可增加成骨诱导的 MEFs 中 Shh 通路因子的表达，从而促进 N-Shh 的成骨效应。此外，IRE1α缺陷的MEFs表现出了成骨标志物水平的升高：我们的研究结果表明，IRE1α介导的未折叠蛋白反应可通过减弱Shh信号传导来缓解MEFs的骨化。我们的研究找到了一种抑制过度骨化的策略，这可能对预防颞下颌关节骨性强直具有临床意义。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Oral diseases 医学-牙科与口腔外科

CiteScore

7.60

自引率

5.30%

发文量

325

审稿时长

4-8 weeks

期刊介绍： Oral Diseases is a multidisciplinary and international journal with a focus on head and neck disorders, edited by leaders in the field, Professor Giovanni Lodi (Editor-in-Chief, Milan, Italy), Professor Stefano Petti (Deputy Editor, Rome, Italy) and Associate Professor Gulshan Sunavala-Dossabhoy (Deputy Editor, Shreveport, LA, USA). The journal is pre-eminent in oral medicine. Oral Diseases specifically strives to link often-isolated areas of dentistry and medicine through broad-based scholarship that includes well-designed and controlled clinical research, analytical epidemiology, and the translation of basic science in pre-clinical studies. The journal typically publishes articles relevant to many related medical specialties including especially dermatology, gastroenterology, hematology, immunology, infectious diseases, neuropsychiatry, oncology and otolaryngology. The essential requirement is that all submitted research is hypothesis-driven, with significant positive and negative results both welcomed. Equal publication emphasis is placed on etiology, pathogenesis, diagnosis, prevention and treatment.