中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2004年
23期
4910-4912
,共3页
金小岚%袁成良%侯建红%钱江龙%黄迎春
金小嵐%袁成良%侯建紅%錢江龍%黃迎春
금소람%원성량%후건홍%전강룡%황영춘
雌二醇/药理学%基因表达%骨髓基质细胞
雌二醇/藥理學%基因錶達%骨髓基質細胞
자이순/약이학%기인표체%골수기질세포
背景:绝经后骨质疏松的病理生理机制尚未完全阐明,研究表明骨丢失与雌激素缺乏相关,后者可导致多种细胞因子生成增加,继而引起成骨细胞和破骨细胞生成增多,骨吸收增强.雌二醇能否改变骨髓基质细胞分化过程中Cbfα1和PPAR-γ2 mRNA的表达进而影响其向成骨细胞分化尚不清楚.目的:研究骨髓基质细胞在向成骨细胞分化的介质中,17β-雌二醇对其核结合因子α1(Cbfα1)及过氧化物酶增殖活化受体γ2(PPAR-γ2)mRNA表达的影响,探讨雌二醇对成骨细胞生成的作用.设计:非随机对照研究.地点和对象:所有实验均在成都军区总医院检验科和成都百奥生物技术有限公司完成,分离骨髓基质细胞所需大鼠由成都中医药大学实验动物研究中心提供.干预:1,25(OH)2D3和地塞米松诱导大鼠骨髓基质细胞向成骨细胞分化,用不同浓度[(0～1)×10-6mol/L]雌二醇对细胞分化过程进行干预.主要观察指标:应用半定量RT-PCR及Northern blot技术,观察不同浓度雌二醇对骨髓基质细胞分化过程中Cbfα1及PPAR-γ2 mRNA表达的影响.结果:雌二醇能明显抑制骨髓基质细胞分化过程中Cbfα1 mRNA的表达,雌二醇浓度为[(0～1)×10-6mol/L]时,Cbfα1 mRNA的表达量从(25.0±3.3)%降至(19.8±2.2)%(t=2.62,P＜0.05),(14.5±1.3)%(t=5.92,P＜0.01)和(6.5±1.9)%(t=9.72,P＜0.01);雌二醇能明显促进骨髓基质细胞在分化介质中PPAR-γ2 mRNA的表达,在上述雌二醇浓度时,PPAR-γ2 mRNA的表达从(1.75±0.5)%增至(9.5±2.1)%(t=7.18,P＜0.01)和(19.3±3.0)%(t=11.5,P＜0.01);Northern blot结果显示随着雌二醇浓度的增加,Cbfα1 mRNA的表达量从4.42±0.39降至1.88±0.16(t=12.0,P＜0.01)和1.19±0.11(t=15.8,P＜0.01);PPAR-γ2mRNA的表达量从1.31±0.12增至2.81±0.22(t=10.5,P＜0.01)和4.02±0.36(t=14.2,P＜0.01).细胞ALP活性明显受雌二醇的抑制,在上述雌二醇浓度时ALP的活性从(42.6±2.5)降至(3.6 ±0.7)U/g蛋白(t=29,P＜0.01).Ⅰ型胶原含量均随雌二醇浓度增加而降低.结论:雌二醇抑制体外培养的骨髓基质细胞向成骨细胞分化而促进其向脂肪细胞分化.
揹景:絕經後骨質疏鬆的病理生理機製尚未完全闡明,研究錶明骨丟失與雌激素缺乏相關,後者可導緻多種細胞因子生成增加,繼而引起成骨細胞和破骨細胞生成增多,骨吸收增彊.雌二醇能否改變骨髓基質細胞分化過程中Cbfα1和PPAR-γ2 mRNA的錶達進而影響其嚮成骨細胞分化尚不清楚.目的:研究骨髓基質細胞在嚮成骨細胞分化的介質中,17β-雌二醇對其覈結閤因子α1(Cbfα1)及過氧化物酶增殖活化受體γ2(PPAR-γ2)mRNA錶達的影響,探討雌二醇對成骨細胞生成的作用.設計:非隨機對照研究.地點和對象:所有實驗均在成都軍區總醫院檢驗科和成都百奧生物技術有限公司完成,分離骨髓基質細胞所需大鼠由成都中醫藥大學實驗動物研究中心提供.榦預:1,25(OH)2D3和地塞米鬆誘導大鼠骨髓基質細胞嚮成骨細胞分化,用不同濃度[(0～1)×10-6mol/L]雌二醇對細胞分化過程進行榦預.主要觀察指標:應用半定量RT-PCR及Northern blot技術,觀察不同濃度雌二醇對骨髓基質細胞分化過程中Cbfα1及PPAR-γ2 mRNA錶達的影響.結果:雌二醇能明顯抑製骨髓基質細胞分化過程中Cbfα1 mRNA的錶達,雌二醇濃度為[(0～1)×10-6mol/L]時,Cbfα1 mRNA的錶達量從(25.0±3.3)%降至(19.8±2.2)%(t=2.62,P＜0.05),(14.5±1.3)%(t=5.92,P＜0.01)和(6.5±1.9)%(t=9.72,P＜0.01);雌二醇能明顯促進骨髓基質細胞在分化介質中PPAR-γ2 mRNA的錶達,在上述雌二醇濃度時,PPAR-γ2 mRNA的錶達從(1.75±0.5)%增至(9.5±2.1)%(t=7.18,P＜0.01)和(19.3±3.0)%(t=11.5,P＜0.01);Northern blot結果顯示隨著雌二醇濃度的增加,Cbfα1 mRNA的錶達量從4.42±0.39降至1.88±0.16(t=12.0,P＜0.01)和1.19±0.11(t=15.8,P＜0.01);PPAR-γ2mRNA的錶達量從1.31±0.12增至2.81±0.22(t=10.5,P＜0.01)和4.02±0.36(t=14.2,P＜0.01).細胞ALP活性明顯受雌二醇的抑製,在上述雌二醇濃度時ALP的活性從(42.6±2.5)降至(3.6 ±0.7)U/g蛋白(t=29,P＜0.01).Ⅰ型膠原含量均隨雌二醇濃度增加而降低.結論:雌二醇抑製體外培養的骨髓基質細胞嚮成骨細胞分化而促進其嚮脂肪細胞分化.
배경:절경후골질소송적병리생리궤제상미완전천명,연구표명골주실여자격소결핍상관,후자가도치다충세포인자생성증가,계이인기성골세포화파골세포생성증다,골흡수증강.자이순능부개변골수기질세포분화과정중Cbfα1화PPAR-γ2 mRNA적표체진이영향기향성골세포분화상불청초.목적:연구골수기질세포재향성골세포분화적개질중,17β-자이순대기핵결합인자α1(Cbfα1)급과양화물매증식활화수체γ2(PPAR-γ2)mRNA표체적영향,탐토자이순대성골세포생성적작용.설계:비수궤대조연구.지점화대상:소유실험균재성도군구총의원검험과화성도백오생물기술유한공사완성,분리골수기질세포소수대서유성도중의약대학실험동물연구중심제공.간예:1,25(OH)2D3화지새미송유도대서골수기질세포향성골세포분화,용불동농도[(0～1)×10-6mol/L]자이순대세포분화과정진행간예.주요관찰지표:응용반정량RT-PCR급Northern blot기술,관찰불동농도자이순대골수기질세포분화과정중Cbfα1급PPAR-γ2 mRNA표체적영향.결과:자이순능명현억제골수기질세포분화과정중Cbfα1 mRNA적표체,자이순농도위[(0～1)×10-6mol/L]시,Cbfα1 mRNA적표체량종(25.0±3.3)%강지(19.8±2.2)%(t=2.62,P＜0.05),(14.5±1.3)%(t=5.92,P＜0.01)화(6.5±1.9)%(t=9.72,P＜0.01);자이순능명현촉진골수기질세포재분화개질중PPAR-γ2 mRNA적표체,재상술자이순농도시,PPAR-γ2 mRNA적표체종(1.75±0.5)%증지(9.5±2.1)%(t=7.18,P＜0.01)화(19.3±3.0)%(t=11.5,P＜0.01);Northern blot결과현시수착자이순농도적증가,Cbfα1 mRNA적표체량종4.42±0.39강지1.88±0.16(t=12.0,P＜0.01)화1.19±0.11(t=15.8,P＜0.01);PPAR-γ2mRNA적표체량종1.31±0.12증지2.81±0.22(t=10.5,P＜0.01)화4.02±0.36(t=14.2,P＜0.01).세포ALP활성명현수자이순적억제,재상술자이순농도시ALP적활성종(42.6±2.5)강지(3.6 ±0.7)U/g단백(t=29,P＜0.01).Ⅰ형효원함량균수자이순농도증가이강저.결론:자이순억제체외배양적골수기질세포향성골세포분화이촉진기향지방세포분화.
BACKGROUND: Bone remodeling rate increases precipitously at menopause, which may be explained that loss of sex steroids up-regulates the formation of osteoclasts and osteoblasts in the marrow by up-regulating the production and action of cytokines responsible for osteoclastogenesis and osteoblastogenesis.However, the molecular mechanism of 17β-estradiol(E2)on osteoblastogenesis is not clear yet.OBJECTIVE: To investigate the effects of 17β-estradiol(E2) on the gene expression of core-binding factor alpha 1 (Cbfα1) and peroxisome proliferator-activated receptor gamma 2(PPAR-γ2) in rat bone marrow stromal cells exposured to the differentiation medium and to elucidate the role of E2 on osteoblastogenesis.DESIGN: A nonrandomized controlled experimental study was conducted.SETTING and PARTICIPANTS: All experiments were carried out in the Department of Laboratory, Chengdu Military Command General Hospital and Chengdu Bai' ao Biol-Tech Limited Company. Three-month-old female SD rat weighing (200 ± 20) g used to isolate bone marrow stromal cells was obtained from the Animal Center of Chengdu University of Traditional Chinese Medicine.INTERVENTION: Adherent bone marrow stromal cells were intervened with dexamethasome (DEX) 1 × 10-7 mol/L, 1, 25(OH)2D3 1 × 10-9 mol/L and different concentrations [ (0 - 1) × 10-6 mol/L] of E2.MAIN OUTCOME MEASURES: The effects of E2 on the Cbfα1 and PPAR-γ2 gene expression were assayed by semiquantitative RT-PCR and demonstrated by Northern blot.RESULTS: A dose dependent inhibition of the expression of Cbfα1 mRNA in bone marrow stromal cell by E2 was discovered. When examined under various concentrations of E2 [ (0 - 1) × 10-6 mol/L], the expression of Cbfα1 mRNA decreased from (25.0 ± 3.3)% to (19.8 ± 2.2)% (t =2.62, P ＜0.05), (14.5±1.3)% (t=5.92, P ＜0.01) and (6.5±1.9)% ( t = 9.72, P ＜ 0. 01 ) . E2 strongly stimulated the expression of PPAR-γ2 mRNA in bone marrow stromal cell from (1.75±0.5)% to (9.5±2.1)%(t=7.18, P ＜0.01) and(19.3±3.0)% (t=11.5, P＜0.01 ) . Northern blot showed that the expression of Cbfα1 and PPAR-γ2 mRNA in bone marrow stromal cell decreased from 4. 42 ± 0. 39 to 1.88 ±0.16(t=12.0, P ＜0.01), (1.19±0.11) (t=15.8, P ＜0.01) and increased from 1.31 ±0. 12 to2. 81 ±0.22 (t = 10.5, P ＜0.01),4.02 ±0.36(t=14.2, P ＜ 0.01), respectively. The activity of Alkaline phoshatase was suppressed from (42.6 ±2.5) U/g to (3.6 ±0. 7) U/gprotein ( t = 29, P ＜ 0. 01 ) by E2. The amount of type Ⅰ collagen decreased significantly at higher concentrations of E2.CONCLUSION: E2 promoted bone marrow stromal cells differentiating into adipocytes and inhibited osteoblastogenesis in vitro.
