摘要
目的 了解骨代谢过程中雌激素与铁对于骨吸收的拮抗效应及氧化应激在其中的作用.方法 建立高铁小鼠模型,随机分为去势组（OVX）、高铁去势组（F＋OVX）和高铁组（F）.检测血清铁蛋白及氧化应激水平,反转录聚合酶链反应（RT-PCR）检测胫骨骨吸收相关基因表达,股骨远端Micro-CT分析.离体实验中用铁剂和雌二醇（E2）干预RAW264.7细胞,观察破骨细胞分化及胞内活性氧物质（ROS）水平.结果 F组与F＋OVX组铁蛋白含量明显升高,分别为（335.3±44.1）μg/L比（41.4±5.6）μg/L,（324.8±38.6） μg/L比（41.4±5.6） μg/L,均P＜0.01;血清丙二醛（MDA）趋势：F＋ OVX组＞OVX组＞F组,超氧化物歧化酶（SOD）趋势与之相反.Micro-CT分析,较之OVX组,F＋ OVX组骨量下降（0.11 ±0.01）mg/mm3比（0.19±0.03）mg/mm^3;F组较F＋OVX组骨量上升（0.90±0.06） mg/mm^3比（0.11±0.01）mg/mm^3,均P ＜0.05;PCR结果,F＋OVX组酒石酸抗酸性磷酸酶（TRAP）及细胞分裂素（CTK）表达较OVX组升高,F组TRAP、转运蛋白（CTR）及CTK表达较F＋ OVX组下降;TRAP染色显示铁剂干预后破骨细胞数量增加（41.7±5.5）比（20.0 ±4.0）,P＜0.05,ROS水平升高（160％±8％）比（100％±9％）,P＜0.05,在此基础上以E2干预后破骨细胞数量下降（14.8±5.1）比（41.7±5.5）,P＜0.05,ROS水平下降（53％±13％）比（160％±8％）,P＜0.05.结论 雌激素与铁对骨吸收存在一定的拮抗效应,该效应可能是通过两者对ROS的调控实现的.
Objective To explore the antagonistic effect of estrogen on iron-induced bone resorption and the role of oxidative stress.Methods In vivo,8-week-old female imprinting control region mice were randomly divided into 3 groups of ferritin （F）,ovariectomy （OVX） and F ＋ OVX.Intervention was made by ferric ammonium citrate （FAC） and OVX.Serum levels of ferritin,malondialdehyde （MDA） and superoxide dismutase （SOD） were measured.The expression changes of TRAP,CTR,matrix metallopeptidase 9 （MMP9） and CTK derived from murine bilateral tibia were detected by reverse transcription-polymerase chain reaction （RT-PCR）.A high-resolution micro-computed tomography was utilized for scanning distal femur.In vitro,RAW264.7 cells were used and intervened by FAC and estradiol.Tartrate resistant acid phosphatase （TRAP） staining was performed and wine-red TRAP positive cells were counted.ROS level was detected by 2＇,7＇-dichloro-dihydrofluorescein diacetate （DCFH-DA） with a multi-detection reader.Results The serum ferritin were heightened in F and F ＋ OVX groups [（335.30 ±44.10） vs （41.38 ±5.56） μg/L,（324.80 ±38.60） vs （41.38 ±5.56） μg/L respectively,P 〈0.01].The trend of MDA level was F ＋ OVX 〉 OVX 〉 F while SOD level was quite opposite.Body mass density of F ＋ OVX group was lower than that of OVX group （0.114 ± 0.013 vs 0.187 ± 0.029 mg/mm^3,P 〈 0.05） or F group （0.114 ± 0.013 vs 0.902 ± 0.064 mg/mm3,P 〈 0.05）.RT-PCR：TRAP and CTK gene expression of OVX group was lower than that of F ＋ OVX group.However,TRAP,CTR and CTK gene expression of F ＋ OVX group was higher than that of F group.TRAP staining：FAC increased the number of TRAP positive cells （41.7 ±5.5 vs 20.0 ±4.0,P 〈0.05） while estradiol decreased it （14.8 ±5.1 vs 41.7 ±5.5,P 〈0.05）.DCFH-DA test show that reactive oxygen species was elevated by FAC （160% ±8% vs 100% ±9%,P 〈0.05） and reduced by estradiol （53% ± 13% vs 160% ±8%,P〈0.05）.Conclusion The antag
出处
《中华医学杂志》
CAS
CSCD
北大核心
2014年第33期2613-2617,共5页
National Medical Journal of China
基金
国家自然科学基金（81273090）
国家自然科学基金青年项目（81302438）
