CAJ | 학술논문

背景:近年来,中药及中药有效部分对骨质疏松的干预和治疗作用的报道较多,但涉及细胞成骨分化调控的信号途径的报道较少. 目的:观察杜仲诱导大鼠骨髓间充质干细胞成骨分化过程中 Wnt 信号途径相关基因表达的变化. 方法:将第3代 SD 大鼠骨髓间充质干细胞,接种到6孔培养板中,每孔1×103个细胞,24 h 后更换诱导培养基(含体积分数为7.5%胎牛血清的 DMEM/F12(1∶1)加1/1000浓度的杜仲醇提取物).阴性对照组仍为正常培养基培养.诱导8 h,1 d,3 d 和7 d 时采用 RT-qPCR 法测定 Wnt 信号途径中 Fzd 和 LRP 受体系列、β-catenin、核内 Wnt 调控靶基因系列及 Wnt 抑制因子(WIF1)等表达变化. 结果与结论:与阴性对照组比较,诱导3 d 后 Fzd2表达升高11.86倍,Fzd3升高达到2倍;诱导7 d 后,Fzd2表达升高5.12倍,Fzd3恢复到正常水平;β-catenin 在诱导3 d 时表达升高达 2倍;WIF1在诱导3 d 和7 d 后表达显著下降.结果提示 Wnt 信号途径可能参与了杜仲促骨髓间充质干细胞成骨分化过程.
배경:근년래,중약급중약유효부분대골질소송적간예화치료작용적보도교다,단섭급세포성골분화조공적신호도경적보도교소. 목적:관찰두중유도대서골수간충질간세포성골분화과정중 Wnt 신호도경상관기인표체적변화. 방법:장제3대 SD 대서골수간충질간세포,접충도6공배양판중,매공1×103개세포,24 h 후경환유도배양기(함체적분수위7.5%태우혈청적 DMEM/F12(1∶1)가1/1000농도적두중순제취물).음성대조조잉위정상배양기배양.유도8 h,1 d,3 d 화7 d 시채용 RT-qPCR 법측정 Wnt 신호도경중 Fzd 화 LRP 수체계렬、β-catenin、핵내 Wnt 조공파기인계렬급 Wnt 억제인자(WIF1)등표체변화. 결과여결론:여음성대조조비교,유도3 d 후 Fzd2표체승고11.86배,Fzd3승고체도2배;유도7 d 후,Fzd2표체승고5.12배,Fzd3회복도정상수평;β-catenin 재유도3 d 시표체승고체 2배;WIF1재유도3 d 화7 d 후표체현저하강.결과제시 Wnt 신호도경가능삼여료두중촉골수간충질간세포성골분화과정.
BACKGROUND: Recently, there are many reports describing Chinese medicine and the effective components of Chinese medicine for treatment of osteoporosis. But there are few reports regarding cel ular osteogenic differentiation and regulation signaling. OBJECTIVE: To investigate the expression of Wnt signaling pathway related genes during osteogenic differentiation of rat bone marrow mesenchymal stem cel s induced by eucommia bark. METHODS: Passage 3 Sprague-Dawley rat bone marrow mesenchymal stem cel s were inoculated into 6-wel plate, with 1×103 cel s per wel . 24 hours later, induced medium supplemented with 7.5% fetal bovine serum-containing DMEM/F12 (1:1) and 1/1 000 eucommia bark was refreshed. Normal culture medium was used in the negative control group. After induction for 8 hours, 1, 3, and 7 days, expression levels of Fzd and LRP receptors of Wnt passway, β-catenin, intranuclear Wnt target genes and Wnt inhibitory factor were determined by RT-qPCR. RESULTS AND CONCLUSION: Compared with negative control group, Fzd2 expression increased to 11.86 folds and Fzd3 expression increased to 2 folds respectively after induction for 3 days; Fzd2 expression increased to 5.12 folds and Fzd3 rebounded to normal level after induction for 7 days; β-catenin expression increased to 2 folds after induction for 3 days, and WIF1 expression decreased significantly after induction for 3 and 7 days, respectively. These findings suggest that Wnt signaling pathway may participate in the process of eucommia bark promoting in osteogenic differentiation of bone marrow mesenchymal stem cel s.