Department of Endocrinology, Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, 215004, China.
Department of Ultrasound, Suzhou Science and Technology Town Hospital, Suzhou, Jiangsu, 215153, China.
Exp Cell Res. 2021 Dec 1;409(1):112890. doi: 10.1016/j.yexcr.2021.112890. Epub 2021 Oct 22.
Formation of osteoclasts is known to be closely associated with osteoporosis progression. LOX is a key enzyme that catalyzes the synthesis of collagen, which is the new mediator in osteoclast formation. However, the effect of LOXG473A on of osteoclast formation needs to be explored. Thereby, we sought to explore the effect of LOX on formation of osteoclasts and its underlying mechanism. To investigate the function of LOX in osteoclast formation, Raw264.7 cells were stably transfected with LOX-WT or LOX-MUT (LOX). Real-time PCR and western blotting were used to detect the relative levels of osteoclast formation related genes and proteins. TRAP staining and immunofluorescence staining were used to test the ability of Raw264.7 cells to form osteoclasts and the ability of cells to form rings, respectively. Bone erosion assay was used to test bone resorptive activity. The data indicated that LOX significantly enhanced the ability of osteoclasts forming, ring-forming and bone resorpting in Raw264.7 cells. Mechanically, LOX upregulated the expressions of NFATC1, ACP5, CTSK, IL-6, and the proportion of p-JAK2/JAK2 and p-STAT3/STAT3, thereby promoting the formation of osteoclasts. In conclusion, we have verified that LOX induces the proliferation of osteoclasts in Raw264.7 cells via IL-6/JAK2/STAT3 signaling, suggesting a novel strategy for studying osteoporosis.
破骨细胞的形成与骨质疏松症的进展密切相关。LOX 是一种催化胶原蛋白合成的关键酶，而胶原蛋白是破骨细胞形成的新介质。然而，LOXG473A 对破骨细胞形成的影响仍需进一步探索。因此，我们旨在探讨 LOX 对破骨细胞形成的影响及其潜在机制。为了研究 LOX 在破骨细胞形成中的功能，我们使用 LOX-WT 或 LOX-MUT（LOX）稳定转染 Raw264.7 细胞。实时 PCR 和 Western blot 用于检测破骨细胞形成相关基因和蛋白的相对水平。TRAP 染色和免疫荧光染色分别用于测试 Raw264.7 细胞形成破骨细胞的能力和细胞形成环的能力。骨侵蚀测定用于测试骨吸收活性。数据表明，LOX 显著增强了 Raw264.7 细胞中破骨细胞形成、环形成和骨吸收的能力。从机制上讲，LOX 上调了 NFATC1、ACP5、CTSK、IL-6 的表达以及 p-JAK2/JAK2 和 p-STAT3/STAT3 的比例，从而促进了破骨细胞的形成。总之，我们已经验证了 LOX 通过 IL-6/JAK2/STAT3 信号通路诱导 Raw264.7 细胞中破骨细胞的增殖，这为研究骨质疏松症提供了一种新策略。
