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-9 mRNA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6000 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。
Objective To investigate the effects of different stages of hypoxia exposure on RANKL/NF-κB/NFATc1pathway 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 6000 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.
作者
张娟
杨历新
王叶
高淑珍
许海琦
樊宇
郭薇薇
ZHANG Juan;YANG Lixin;WANG Ye;GAO Shuzhen;XU Haiqi;FAN Yu;GUO Weiwei(Qinghai University Graduate School,Xining 810000,China;Endocrinology Department of Qinghai Provincial People’s Hospital,Xining 810000,China)
出处
《中国骨质疏松杂志》
北大核心
2025年第8期1121-1127,共7页
Chinese Journal of Osteoporosis
基金
2022年青海省“昆仑英才·高端创新创业人才”项目(QHKLYC-GDCXCY-2022-048)。
