中国骨质疏松杂志

低氧暴露对小鼠RANKL/NF‐κB/NFATc1通路影响及骨代谢变化

Effects of hypoxia exposure on RANKL/NF-κB/NFATc1 pathway and changes in bone metabolism in mice

DOI：10.3969/j.issn.1006-7108.2025.08.005

英文关键词:hypoxia osteoporosis osteoclast NF-κB NFATc1

基金项目:2022年青海省“昆仑英才·高端创新创业人才”项目（QHKLYC-GDCXCY-2022-048）

作者	单位
张娟1 杨历新2* 王叶2 高淑珍1 樊宇1 郭薇薇1	1.青海大学研究生院，青海西宁810000 2.青海省人民医院内分泌科，青海西宁810000

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中文摘要:

目的探讨低氧暴露不同阶段对小鼠RANKL/NF‐κB/NFATc1通路影响及骨代谢变化。方法将48只SPF级C57BL/6J雄性小鼠随机分为低氧组（hypoxia，H组）和对照组（control，C组），24只/组，低氧组小鼠在低压氧舱中构建缺氧模型，进一步根据时间依赖性分为H1、H5、H14、H28四个亚组，同时为每个时间段设置对照组C1、C5、C14、C28。观察并记录各组小鼠的体重变化，通过ELISA检测IL-6、TNF-a及PINP、CTX-I水平；HE染色观察骨组织病理变化；qRT-PCR检测促破骨细胞分化基因包括ACP5、CTSK、MMP-9的表达；Western Blot检测HIF-1a及RANKL、NF‐κB、NFATc1蛋白的表达。结果低氧组小鼠体重均低于对照组（P＜0.05），且低氧暴露1、5、14、28 d分别较基础体重下降7.27%、8.67%、7.32%、18.78%；低氧暴露14 d后出现骨小梁数量、体积减少，连接稀疏等情况，28 d后这种改变进一步增加；低氧暴露增加了HIF-1a、RANKL、NF-κB、NFATc1蛋白的表达，与对照组相比，H5、H28组蛋白表达更显著（P＜0.05）；低氧促进了ACP5、CTSK、MMP-9mRNA的表达,且H14、H28组分别高于C14、C28组（P＜0.05）；低氧暴露导致IL-6、TNF-a、PINP、CTX-I水平升高，IL-6、TNF-a低氧组均高于对照组（P＜0.05），PINP水平H5、H14、H28组分别高于C5、C14、C28组（P＜0.05），CTX-I水平H5、H28组分别高于C5、C28组（P＜0.05）。结论模拟海拔6 000 m环境下，低氧暴露使小鼠应激性消耗减重，通过上调RANKL/NF‐κB/NFATc1信号通路和促进炎症导致骨代谢活跃及骨质流失。

英文摘要:

Objective To investigate the effects of different stages of hypoxia exposure on RANKL/NF-κB/NFATc1 pathway and bone metabolism in mice. Methods A total of 48 SPF C57BL/6J male mice were randomly divided into hypoxia group (H group) and control group (Cgroup), 24 in each group. The mice in the hypoxia group were constructed in a low-pressure oxygen chamber. The hypoxia model was further divided into four subgroups H1, H5, H14, and H28 in a time-dependent manner, and the control groups C1, C5, C14, and C28 were set for each time period. The body weight changes of mice in each group were observed and recorded. The levels of IL-6, TNF-a, PINP and CTX-I were detected by ELISA.HE staining was used to observe the pathological changes of bone tissue. The expression of osteoclast differentiation genes including ACP5, CTSK and MMP-9 was detected by qRT-PCR. Western Blot was used to detect the expression of HIF-1a, RANKL, NF-κB and NFATc1 proteins. Results The body weightof mice in the hypoxia group was lower than that in the control group (P<0.05), and the body weight ofmice exposed to hypoxia for 1 day, 5 days, 14 days and 28 days decreased by 7.27 %, 8.67 %, 7.32 % and 18.78 %, respectively. After 14 days of hypoxia exposure, the number and volume of trabecular bonedecreased, and the connection was sparse. After 28 days, this change further increased. Hypoxia exposure increased the expression of HIF-1a, RANKL, NF-κB and NFATc1 proteins. Compared with the control group, the expression of H5 and H28 proteins was more significant (P<0.05). Hypoxia promoted the expression of ACP5, CTSK and MMP-9 mRNA, and the H14 and H28 groups were higher than the C14and C28 groups (P<0.05). Hypoxia exposure increased the levels of IL-6, TNF-a, PINP and CTX-I. Thelevels of IL-6 and TNF-a in hypoxia group were higher than those in control group (P<0.05). The levels of PINP in H5, H14 and H28 groups were higher than those in C5, C14 and C28 groups (P<0.05). The levels of CTX-I in H5 and H28 groups were higher than those in C5 and C28 groups (P<0.05). Conclusion Under the simulated altitude of 6 000 m, hypoxia exposure reduced the stress consumption of mice, and promoted bone metabolism and bone loss by up-regulating RANKL/NF-κB/NFATc1 signaling pathway and promoting inflammation.

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