CAJ | 학술논문

目的观察WNT信号通路激活剂SB-216763对于人骨髓基质于细胞(hBMSCs)成脂-成骨分化的影响.方法从3例临床股骨头标本中分离培养hBMSCs与5μmol/L SB-216763共同培养,观察hBMSCs表达β-连锁蛋白(β-catenin)的变化,检测hBMSCs增殖能力、成脂分化能力和成骨细胞诱导分化时碱性磷酸酶(ALP)活性的变化.结果 SB-216763可以显著促进hBMSCs表达β-catenin (30.2±2.7比13.3±2.1,P＜0.01)、促进hBMSCs增殖(P＜0.01)、抑制hBMSCs向脂肪细胞分化[油红O(Oil Red-O)阳性细胞数:27.4±3.2比8.4±2.1,P＜0.01]、降低hBMSCs的ALP活性(P＜0.01).结论激活WNT信号通路可以促进hBMSCs的增殖,抑制hBMSCs向脂肪细胞分化并且抑制hBMSCs的成骨活性.
목적 관찰WNT신호통로격활제SB-216763대우인골수기질우세포(hBMSCs)성지-성골분화적영향.방법 종3례림상고골두표본중분리배양hBMSCs여5μmol/L SB-216763공동배양,관찰hBMSCs표체β-련쇄단백(β-catenin)적변화,검측hBMSCs증식능력、성지분화능력화성골세포유도분화시감성린산매(ALP)활성적변화.결과 SB-216763가이현저촉진hBMSCs표체β-catenin (30.2±2.7비13.3±2.1,P＜0.01)、촉진hBMSCs증식(P＜0.01)、억제hBMSCs향지방세포분화[유홍O(Oil Red-O)양성세포수:27.4±3.2비8.4±2.1,P＜0.01]、강저hBMSCs적ALP활성(P＜0.01).결론 격활WNT신호통로가이촉진hBMSCs적증식,억제hBMSCs향지방세포분화병차억제hBMSCs적성골활성.
Objective To investigate the effects of a WNT signaling pathway activator,SB-216763,on the adipocytogenesis and osteocytogenesis of human bone marrow stromal cells (hBMSCs) in vitro.Methods hBMSCs were isolated and cultured from 3 cases of femoral head specimens.Upon 60％confluence of hBMSCs,medium was changed to DMEM containing 5 μmol/l SB-216763.After 24 h,the expression of β-catenin was detected by using immunocytochemistry.Methyl thiazol tetrazolium (MTT) assay was performed to analyze the proliferation of hBMSCs after the cells were cultured in 96-well plate with 5 μmol/L SB-216763 for 72 h.Upon confluence of hBMSCs,medium was changed to DMEM containing adipocytogenic or osteocytogenic supplements in the presence or absence of 5 μmol/L SB-216763.After 10 days,the adipocytogenic cultures were stained with Oil Red-O and the number of Oil Red-O positive cells was counted. For osteocytogenic cultures,the ALP activity was evaluated after 7 days.Results SB216763 treatment produced more nuclear expression positive cells (30.2 ± 2.7 ) than the control group (13.3 ±2.1 ) ( P ＜0.01 ),and stimulated the proliferation of hBMSCs determined by MTT assay ( P ＜0.01).Generation of Oil Red-O-positive cells was inhibited significantly by SB-216763 (27.4 ± 3.2 vs.8.4 ±2.1,P＜0.01).ALP activity was inhibited by SB-216763 in hBMSCs under osteocytogenesis ( P ＜0.01 ).Conclusion The WNT signaling pathway activator,SB-216763 stimulates proliferation of hBMSCs,and inhibits both adipocytogenesis and osteocytogenesis of hBMSCs.