铁蓄积对老年雌性小鼠骨代谢影响的实验研究  

Experimental research of the impact of iron accumulation on bone metabolism in elderly female mice

作　　者：朱胤 刘中瑞 刘桂文 韩皓衍 肖春林 肖建华 叶勇军 黄为民 刘午阳 刘禄林 ZHU Yin;LIU Zhong-rui;LIU Gui-wen;HAN Hao-yan;XIAO Chun-lin;XIAO Jian-hua;YE Yong-jun;HUANG Wei-min;LIU Wu-yang;LIU Lu-lin(The First Clinical Medical School of Gannan Medical University;Department of Orthopedics,The First Affiliated Hospital of Gannan Medical University;Ganzhou Key Laboratory of Osteoporosis Research,Ganzhou,Jiangxi 341000)

机构地区：[1]赣南医科大学第一临床医学院 [2]赣南医科大学第一附属医院骨科 [3]赣州市骨质疏松研究重点实验室,江西赣州341000

出　　处：《赣南医科大学学报》2025年第3期211-217,共7页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：国家自然科学基金地区项目(82260649);江西省自然科学基金资助项目(20232BAB216028)。

摘　　要：目的:通过敲除铁调素基因(Hepcidin 1,Hepc1)构建老年雌性小鼠内源性铁蓄积模型,探究慢性铁蓄积对老年雌性小鼠骨代谢的作用及相应机制。方法:将Hepc1杂合子小鼠交配繁殖后,根据子代基因鉴定结果将小鼠分为2组:野生型(Wlid type,WT)小鼠组、Hepc1基因敲除型(Knockout,KO)小鼠组,每组6只。小鼠18月龄时为实验研究终点,采用原子吸收光谱法和Perl's染色法检测铁蓄积状况。通过ELISA法检测血清雌二醇、铁调素、铁蛋白、骨转换标志物浓度,Micro-CT观察骨微结构,q-PCR测定成骨和破骨相关基因的表达,化学比色法检测氧化应激水平。结果:2组小鼠的体重和雌激素水平差异无统计学意义(P>0.05),KO组血清铁调素浓度显著低于WT组(P<0.001)。与WT组相比,KO组血清铁蛋白浓度、肝脏和骨铁含量显著升高(P<0.001)。Perl's染色显示KO组肝脏有典型的铁沉积表现。Micro-CT结果显示,与WT组相比,KO组松质骨的骨密度、骨小梁体积分数、骨小梁数量、骨小梁厚度、连接密度显著降低(P<0.05),骨小梁分离度和结构模型指数显著升高(P<0.05),皮质骨面积显著减小(P<0.05)。与WT组相比,KO组破骨相关基因CTSK、MMP9、OPG、TRAP、RANKL表达以及Ⅰ型胶原羧基末端肽(C-terminal telopeptide of typeⅠcollagen,CTX)水平上升(P<0.05),成骨相关基因ALP、COL-1、OCN、RUNX2、SP7表达以及骨钙素(Osteocalcin,OCN)水平下降(P<0.05)。与WT组相比,KO组抗氧化指标超氧化物歧化酶(Superoxide dismutase,SOD)水平下降和氧化指标丙二醛(Malondialdehyde,MDA)水平上升(P<0.001)。结论:小鼠Hepc1基因敲除后表现为慢性铁蓄积,铁蓄积通过抑制成骨、促进破骨活动加剧骨量丢失,其机制可能与活性氧水平升高有关。Objective:To investigate the effects of chronic iron accumulation on bone microstructure,osteogenesis,and osteoclastogenesis in aged female mice,an endogenous iron accumulation model was developed by knocking out the Hepcidin 1(Hepc1)gene.The study also aimed to clarify the underlying mechanisms of these effects.Methods:Hepc1 heterozygous mice were bred,and offspring were divided into two groups based on genotyping:wild-type(WT)group and Hepc1 knockout(KO)group,with six mice in each group.At the age of 18 months,the mice were sacrificed,and atomic absorption spectrometry along with Perl's staining were employed to evaluate and compare the iron accumulation status between the two groups of mice.ELISA was used to detect the levels of estradiol,iron metabolism markers and bone turnover markers in the serum of mice.The bone microstructure was observed by Micro-CT.q-PCR was used to determine the expression of genes related to bone formation and bone resorption.The level of oxidative stress was detected by chemical colorimetry.Results:There was no significant difference in body weight and estradiol levels between the two groups(P>0.05).Compared with the WT group,the hepcidin content in the KO group was significantly decreased(P<0.001),while the contents of serum ferritin,liver and tibial iron were significantly increased(P<0.001),and Perl's staining also confirmed that there was iron deposition in the liver tissue of the KO group.Compared with the WT group,done mineral density,trabecular thickness,bone volume fraction,trabecular number and connectivity density in the KO group were significantly decreased(P<0.05),and trabecular separation and structure model index were significantly increased(P<0.05);the cortical bone area also decreased(P<0.05).The expressions of osteoclast-related genes such as CTSK,MMP9,OPG,TRAP and RANKL and the level of C-terminal telopeptide of typeⅠcollagen(CTX)in the KO group were higher than those in the WT group(P<0.05),while the expressions of osteogenesis-related genes such as ALP,COL-1,OC

关 键 词：铁调素 铁蓄积 骨转换 氧化应激 

分 类 号：K336[历史地理—历史学]