CAJ | 학술논문

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/mm3;F组较F+OVX组骨量上升(0.90±0.06) mg/mm3比(0.11±0.01)mg/mm3,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.
목적 료해골대사과정중자격소여철대우골흡수적길항효응급양화응격재기중적작용.방법 건립고철소서모형,수궤분위거세조(OVX)、고철거세조(F+OVX)화고철조(F).검측혈청철단백급양화응격수평,반전록취합매련반응(RT-PCR)검측경골골흡수상관기인표체,고골원단Micro-CT분석.리체실험중용철제화자이순(E2)간예RAW264.7세포,관찰파골세포분화급포내활성양물질(ROS)수평.결과 F조여F+OVX조철단백함량명현승고,분별위(335.3±44.1)μg/L비(41.4±5.6)μg/L,(324.8±38.6) μg/L비(41.4±5.6) μg/L,균P＜0.01;혈청병이철(MDA)추세:F+ OVX조＞OVX조＞F조,초양화물기화매(SOD)추세여지상반.Micro-CT분석,교지OVX조,F+ OVX조골량하강(0.11 ±0.01)mg/mm3비(0.19±0.03)mg/mm3;F조교F+OVX조골량상승(0.90±0.06) mg/mm3비(0.11±0.01)mg/mm3,균P ＜0.05;PCR결과,F+OVX조주석산항산성린산매(TRAP)급세포분렬소(CTK)표체교OVX조승고,F조TRAP、전운단백(CTR)급CTK표체교F+ OVX조하강;TRAP염색현시철제간예후파골세포수량증가(41.7±5.5)비(20.0 ±4.0),P＜0.05,ROS수평승고(160％±8％)비(100％±9％),P＜0.05,재차기출상이E2간예후파골세포수량하강(14.8±5.1)비(41.7±5.5),P＜0.05,ROS수평하강(53％±13％)비(160％±8％),P＜0.05.결론 자격소여철대골흡수존재일정적길항효응,해효응가능시통과량자대ROS적조공실현적.
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/mm3,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 antagonistic effect of estrogen on iron-induced bone resorption is probably regulated by oxidative stress.