中华骨科杂志
中華骨科雜誌
중화골과잡지
Chinese Journal of Orthopaedics
2015年
10期
1011-1016
,共6页
王啸%刘禄林%沈光思%费蓓蓓%汪升%姜宇%林华%徐又佳
王嘯%劉祿林%瀋光思%費蓓蓓%汪升%薑宇%林華%徐又佳
왕소%류록림%침광사%비배배%왕승%강우%림화%서우가
雌二醇%骨质疏松%成骨细胞
雌二醇%骨質疏鬆%成骨細胞
자이순%골질소송%성골세포
Estradiol%Osteoporosis%Osteoblasts
目的 探讨单纯雌激素下降和雌激素下降+铁蓄积两种状况下,小鼠骨组织骨量、成骨活性指标、原代成骨细胞代谢指标的差异.方法 第一部分:将体重(25±1)g的ICR小鼠按体重区组随机化分组分为3组,每组7只:(1)去势组,模拟绝经状态;(2)高铁去势组,模拟绝经状态合并铁蓄积,该组小鼠采用“去势”后行枸橼酸铁铵腹腔注射;(3)假手术组.12周后检测指标:小鼠体重、血清铁蛋白;胫骨组织的成骨特异性转录因子(Runx2、Osterix)、骨钙蛋白(Bglap)、骨保护素、雌激素受体α、β(ER-α、ER-β)基因表达;Micro-CT分析股骨远端骨量差异.第二部分:提取新生ICR小鼠颅骨原代成骨细胞,用枸橼酸铁胺、17β-雌二醇(E2)腹腔注射干预,碱性磷酸酶染色观察成骨细胞形态,定量PCR(q-PCR)检测成骨相关基因表达.结果 各组小鼠体重无统计学差异;血清铁蛋白含量:高铁去势组[(324.82±38.60) μg/L]较去势组[(41.38±5.56)μg/L]及假手术组[(39.40±3.81) μg/L]升高,差异有统计学意义(2) Micro-CT的骨密度:高铁去势组[(0.902±0.064) mg/mm3]及去势组[(0.114±0.013)mg/mm3]较假手术组[(0.297±0.072) mg/mm3]均下降,差异有统计学意义,高铁去势组比去势组下降更显著.(3)RT-PCR:与假手术组相比,去势组骨保护素、Runx2、Osterix、Bglap、ER-β表达降低;高铁去势组Osterix、骨保护素表达更低.(4)成骨细胞碱性磷酸酶染色显示用枸橼酸铁胺干预后细胞变小,棕色变淡;q-PCR结果显示:E2升高Bglap表达,枸橼酸铁胺下调Runx2,Osterix表达水平.结论 当雌激素去除时,如合并铁增加,则成骨活性指标进一步下调、骨形成进一步受到抑制.
目的 探討單純雌激素下降和雌激素下降+鐵蓄積兩種狀況下,小鼠骨組織骨量、成骨活性指標、原代成骨細胞代謝指標的差異.方法 第一部分:將體重(25±1)g的ICR小鼠按體重區組隨機化分組分為3組,每組7隻:(1)去勢組,模擬絕經狀態;(2)高鐵去勢組,模擬絕經狀態閤併鐵蓄積,該組小鼠採用“去勢”後行枸櫞痠鐵銨腹腔註射;(3)假手術組.12週後檢測指標:小鼠體重、血清鐵蛋白;脛骨組織的成骨特異性轉錄因子(Runx2、Osterix)、骨鈣蛋白(Bglap)、骨保護素、雌激素受體α、β(ER-α、ER-β)基因錶達;Micro-CT分析股骨遠耑骨量差異.第二部分:提取新生ICR小鼠顱骨原代成骨細胞,用枸櫞痠鐵胺、17β-雌二醇(E2)腹腔註射榦預,堿性燐痠酶染色觀察成骨細胞形態,定量PCR(q-PCR)檢測成骨相關基因錶達.結果 各組小鼠體重無統計學差異;血清鐵蛋白含量:高鐵去勢組[(324.82±38.60) μg/L]較去勢組[(41.38±5.56)μg/L]及假手術組[(39.40±3.81) μg/L]升高,差異有統計學意義(2) Micro-CT的骨密度:高鐵去勢組[(0.902±0.064) mg/mm3]及去勢組[(0.114±0.013)mg/mm3]較假手術組[(0.297±0.072) mg/mm3]均下降,差異有統計學意義,高鐵去勢組比去勢組下降更顯著.(3)RT-PCR:與假手術組相比,去勢組骨保護素、Runx2、Osterix、Bglap、ER-β錶達降低;高鐵去勢組Osterix、骨保護素錶達更低.(4)成骨細胞堿性燐痠酶染色顯示用枸櫞痠鐵胺榦預後細胞變小,棕色變淡;q-PCR結果顯示:E2升高Bglap錶達,枸櫞痠鐵胺下調Runx2,Osterix錶達水平.結論 噹雌激素去除時,如閤併鐵增加,則成骨活性指標進一步下調、骨形成進一步受到抑製.
목적 탐토단순자격소하강화자격소하강+철축적량충상황하,소서골조직골량、성골활성지표、원대성골세포대사지표적차이.방법 제일부분:장체중(25±1)g적ICR소서안체중구조수궤화분조분위3조,매조7지:(1)거세조,모의절경상태;(2)고철거세조,모의절경상태합병철축적,해조소서채용“거세”후행구연산철안복강주사;(3)가수술조.12주후검측지표:소서체중、혈청철단백;경골조직적성골특이성전록인자(Runx2、Osterix)、골개단백(Bglap)、골보호소、자격소수체α、β(ER-α、ER-β)기인표체;Micro-CT분석고골원단골량차이.제이부분:제취신생ICR소서로골원대성골세포,용구연산철알、17β-자이순(E2)복강주사간예,감성린산매염색관찰성골세포형태,정량PCR(q-PCR)검측성골상관기인표체.결과 각조소서체중무통계학차이;혈청철단백함량:고철거세조[(324.82±38.60) μg/L]교거세조[(41.38±5.56)μg/L]급가수술조[(39.40±3.81) μg/L]승고,차이유통계학의의(2) Micro-CT적골밀도:고철거세조[(0.902±0.064) mg/mm3]급거세조[(0.114±0.013)mg/mm3]교가수술조[(0.297±0.072) mg/mm3]균하강,차이유통계학의의,고철거세조비거세조하강경현저.(3)RT-PCR:여가수술조상비,거세조골보호소、Runx2、Osterix、Bglap、ER-β표체강저;고철거세조Osterix、골보호소표체경저.(4)성골세포감성린산매염색현시용구연산철알간예후세포변소,종색변담;q-PCR결과현시:E2승고Bglap표체,구연산철알하조Runx2,Osterix표체수평.결론 당자격소거제시,여합병철증가,칙성골활성지표진일보하조、골형성진일보수도억제.
Objective To explore the effect of iron on bone formation with or without estradiol.Methods 8-week old female ICR mice 25±1 g were randomly divided into 3 groups (n=7 each):SHAM group,OVX group and F+OVX group,which were intervened by ferric ammonium citrate (FAC) with a dose of 0.10 g/kg per week.After 8 weeks,the mice of OVX group and F+ OVX group were spayed (with ovariotomy).12 weeks later,body weight,serum ferritin were measured.The expression changes of Runx2,Osterix,Bglap,OPG,ER-α,ER-β derived from mice tibiae were detected by RT-PCR.A high resolution Micro-CT was used to scan the femur.Primary osteoblasts,extracted from suckling mice' s cranium,were intervened by FAC and estradiol.Har-vested cells after 24 h,then operated alkaline phosphatase staining and q-PCR.Gene expressions of Runx2,osterix and Bglap were measured.Results There were no significant difference between every group in body weight The serum Fer was heightened in F+OVX group (324.82±38.60 μg/L) comparing with SHAM group (39.40±3.81 μg/L) and OVX group (41.38±5.56 μg/L).The differences were statistically significant among the three groups.BMD of O VX group (0.114±0.013 mg/mm3) was lower than that of SHAM group (0.297±0.072 mg/mm3),but higher than that of F+OVX group (0.902±0.064 mg/mm3),The differences were statistically significant among the three groups.RT-PCR results:The expressions of OPG,Runx2,osterix,Bglap and ER-β were decreased after ovariotomy,while the level of osterix and OPG dropped further with the treatment of FAC.In vitro experiment,results of ALP staining suggested that FAC decrease the level of ALP.Q-PCR showed that the expressions of Runx2,osterix were reduced by FAC,while estradiol increased Bglap expression.Conclusion Iron accumulation might have further result in inhibition of bone formation and suppression of osteogenic activity without estradiol.