中国骨质疏松杂志
中國骨質疏鬆雜誌
중국골질소송잡지
CHINESE JOURNAL OF OSTEOPOROSIS
2014年
4期
347-352
,共6页
史淇月%杨柳%魏铂沅%范金柱%王龙%颉强%刘建%罗卓荆
史淇月%楊柳%魏鉑沅%範金柱%王龍%頡彊%劉建%囉卓荊
사기월%양류%위박원%범금주%왕룡%힐강%류건%라탁형
雌激素%破骨细胞%EphB4/EphrinB2
雌激素%破骨細胞%EphB4/EphrinB2
자격소%파골세포%EphB4/EphrinB2
Estrogen%Ostoecalst%EphB4/EphrinB2
目的：研究雌激素通过调节EphB4/EphrinB2信号通路对破骨细胞分化的影响。方法（1）采用RANKL诱导法培养卵巢摘除骨质疏松模型（OVX）组和假手术（Sham）组小鼠的破骨细胞，于第10天提取两组细胞的RNA和蛋白质样品，通过RT-PCR和Westernblot检测细胞EphrinB2表达的变化；（2）采用雌激素及雌激素拮抗剂分别诱导培养RAW264.7破骨细胞，培养第8天提取RNA和蛋白质样品，利用RT-PCR和Westernblot检测细胞EphrinB2表达的变化；（3）在OVX模型组小鼠的破骨细胞培养中添加EphrinB2配体EphB4-Fc片段，通过RT-PCR检测破骨细胞标志物的表达和抗酒石酸酸性磷酸酶（TRAP）染色并计数，观察破骨细胞分化能力的变化。结果卵巢摘除骨质疏松模型组小鼠EphrinB2的表达量低于假手术组（P＜0.01）；雌激素拮抗剂组EphrinB2的表达量低于对照组（P＜0.01），而雌激素组EphrinB2的表达量高于对照组（P＜0.05）；给予EphrinB2的配体EphB4-Fc片段后，OVX组小鼠的破骨细胞标志物表达量降低（P＜0.01，P＜0.001），TRAP染色阳性细胞数减少。结论雌激素可以通过调节破骨细胞EphrinB2的表达影响破骨细胞分化，采用EphB4-Fc片段处理后，OVX组小鼠增强的破骨细胞分化受到抑制。
目的：研究雌激素通過調節EphB4/EphrinB2信號通路對破骨細胞分化的影響。方法（1）採用RANKL誘導法培養卵巢摘除骨質疏鬆模型（OVX）組和假手術（Sham）組小鼠的破骨細胞，于第10天提取兩組細胞的RNA和蛋白質樣品，通過RT-PCR和Westernblot檢測細胞EphrinB2錶達的變化；（2）採用雌激素及雌激素拮抗劑分彆誘導培養RAW264.7破骨細胞，培養第8天提取RNA和蛋白質樣品，利用RT-PCR和Westernblot檢測細胞EphrinB2錶達的變化；（3）在OVX模型組小鼠的破骨細胞培養中添加EphrinB2配體EphB4-Fc片段，通過RT-PCR檢測破骨細胞標誌物的錶達和抗酒石痠痠性燐痠酶（TRAP）染色併計數，觀察破骨細胞分化能力的變化。結果卵巢摘除骨質疏鬆模型組小鼠EphrinB2的錶達量低于假手術組（P＜0.01）；雌激素拮抗劑組EphrinB2的錶達量低于對照組（P＜0.01），而雌激素組EphrinB2的錶達量高于對照組（P＜0.05）；給予EphrinB2的配體EphB4-Fc片段後，OVX組小鼠的破骨細胞標誌物錶達量降低（P＜0.01，P＜0.001），TRAP染色暘性細胞數減少。結論雌激素可以通過調節破骨細胞EphrinB2的錶達影響破骨細胞分化，採用EphB4-Fc片段處理後，OVX組小鼠增彊的破骨細胞分化受到抑製。
목적：연구자격소통과조절EphB4/EphrinB2신호통로대파골세포분화적영향。방법（1）채용RANKL유도법배양란소적제골질소송모형（OVX）조화가수술（Sham）조소서적파골세포，우제10천제취량조세포적RNA화단백질양품，통과RT-PCR화Westernblot검측세포EphrinB2표체적변화；（2）채용자격소급자격소길항제분별유도배양RAW264.7파골세포，배양제8천제취RNA화단백질양품，이용RT-PCR화Westernblot검측세포EphrinB2표체적변화；（3）재OVX모형조소서적파골세포배양중첨가EphrinB2배체EphB4-Fc편단，통과RT-PCR검측파골세포표지물적표체화항주석산산성린산매（TRAP）염색병계수，관찰파골세포분화능력적변화。결과란소적제골질소송모형조소서EphrinB2적표체량저우가수술조（P＜0.01）；자격소길항제조EphrinB2적표체량저우대조조（P＜0.01），이자격소조EphrinB2적표체량고우대조조（P＜0.05）；급여EphrinB2적배체EphB4-Fc편단후，OVX조소서적파골세포표지물표체량강저（P＜0.01，P＜0.001），TRAP염색양성세포수감소。결론자격소가이통과조절파골세포EphrinB2적표체영향파골세포분화，채용EphB4-Fc편단처리후，OVX조소서증강적파골세포분화수도억제。
Objective To investigate the effect of estrogen on osteoclast differentiation by regulating EphB4/EphrinB2 signaling pathway.Methods Osteoclasts of mice in the OVX group and sham group were cultured in the presence of RANKL.The extraction of mRNA and protein in both groups was performed after 10-day culture.The expression of EphrinB2 was detected using RT-PCR and Western boltting.RAW264.7 cells were cultured and induced with estrogen ( E2 group) and estrogen ( ICI group) antagonist, respectively.At the 8th day, the extraction of mRNA and protein was performed.The expression of EphrinB2 was also detected using RT-PCR and Western blotting.The osteoclasts of mice in the OVX group were cultured in the presence of IgG-Fc or EphB4-Fc fragment, a ligand of EphrinB2.The expression of osteoclast markers was detected using RT-PCR.Simultaneously, TRAP staining and counting was performed, in order to observe the osteoclast differentiation.Results The expression of EphrinB2 in OVX group was significantly lower than that in sham group ( P<0.01 ) .Compared with that in control group, the expression of EphrinB2 increased in E2 group but decreased in ICI group ( P<0.01; P<0.05 ) .The intervention of EphB4-Fc down-regulated the expression of osteoclast markers in OVX group (P<0.01;P<0.001), and the counts of TRAP positive cells also decreased.Conclusion Estrogen can influence osteoclast differentiation by regulating the expression of EphrinB2 in osteoclasts.After the intervention of EphB4-Fc, the enhanced osteoclasts differentiation in OVX mice can be suppressed.
