CAJ | 학술논문

背景：ERK1/2信号通路和核因子κB信号通路是否参与了牵张应力作用下MC3T3-E1细胞成骨分化及相关基因表达的调控，尚不清楚。 　　目的：观察机械牵张应力对作用下ERK1/2和核因子κB通路对成骨细胞碱性磷酸酶、Ⅰ型胶原、骨钙蛋白、白细胞介素6表达的影响，探讨ERK1/2与核因子κB信号通路对成骨细胞分化的调控作用。 　　方法：体外培养的MC3T3-E1细胞，以ERK1/2通路特异性抑制剂PD098059及核因子κB通路抑制剂PDTC分别处理30 min后加载12%的拉伸应变率24 h，以正常细胞及单纯加载12%牵张应力24 h为对照。采用ELISA及Real-time PCR方法检测细胞加载前后碱性磷酸酶活性、Ⅰ型胶原、骨钙蛋白及白细胞介素6 mRNA的表达。 　　结果与结论：在12%牵张应力作用下，MC3T3-E1细胞碱性磷酸酶、Ⅰ型胶原、白细胞介素6的表达受ERK1/2信号通路的调控，而骨钙蛋白基因表达的变化不受 ERK1/2通路的影响。核因子κB 信号通路抑制剂 PDTC可显著抑制机械牵应张力作用下MC3T3-E1细胞碱性磷酸酶活性的降低，同时抑制白细胞介素6基因的表达，而Ⅰ型胶原、骨钙蛋白基因表达的变化不受核因子κB信号通路的影响。结果表明牵张应力可以通过ERK1/2和核因子κB通路影响MC3T3-E1细胞的成骨分化及相关基因表达。
배경：ERK1/2신호통로화핵인자κB신호통로시부삼여료견장응력작용하MC3T3-E1세포성골분화급상관기인표체적조공，상불청초。 　　목적：관찰궤계견장응력대작용하ERK1/2화핵인자κB통로대성골세포감성린산매、Ⅰ형효원、골개단백、백세포개소6표체적영향，탐토ERK1/2여핵인자κB신호통로대성골세포분화적조공작용。 　　방법：체외배양적MC3T3-E1세포，이ERK1/2통로특이성억제제PD098059급핵인자κB통로억제제PDTC분별처리30 min후가재12%적랍신응변솔24 h，이정상세포급단순가재12%견장응력24 h위대조。채용ELISA급Real-time PCR방법검측세포가재전후감성린산매활성、Ⅰ형효원、골개단백급백세포개소6 mRNA적표체。 　　결과여결론：재12%견장응력작용하，MC3T3-E1세포감성린산매、Ⅰ형효원、백세포개소6적표체수ERK1/2신호통로적조공，이골개단백기인표체적변화불수 ERK1/2통로적영향。핵인자κB 신호통로억제제 PDTC가현저억제궤계견응장력작용하MC3T3-E1세포감성린산매활성적강저，동시억제백세포개소6기인적표체，이Ⅰ형효원、골개단백기인표체적변화불수핵인자κB신호통로적영향。결과표명견장응력가이통과ERK1/2화핵인자κB통로영향MC3T3-E1세포적성골분화급상관기인표체。
BACKGROUND:The regulatory role of extracellular signal regulated kinase 1/2 (ERK1/2) and nuclear factor kappa B (NF-κB) signal pathways in the osteogenic differentiation of MC3T3-E1 cells subjected to mechanical strain remains unclear. OBJECTIVE:To investigate the effects of ERK1/2 and NF-kB signal pathway on alkaline phosphatase, type Ⅰcol agen, osteocalcin and interleukin-6 expression in osteoblasts in response to mechanical strain, and to explore the regulatory effects of ERK1/2 and NF-kB signal pathway on osteoblast differentiation. METHODS:MC3T3-E1 cells cultured in vitro were separately treated with ERK1/2 pathway specific inhibitor PD098059 and NF-kB pathway inhibitor PDTC for 30 minutes, and subjected to12%elongation for 24 hours. Normal cells and cells along loading 12%mechanical strain for 24 hours were considered as controls. Enzyme linked immunosorbent assay and real-time PCR were utilized to detect alkaline phosphatase activities, type Ⅰcol agen, osteocalcin and interleukin-6 mRNA expression before and after cellloading. RESULTS AND CONCLUSION:Under 12%mechanical strain, alkaline phosphatase, type I col agen, and interleukin-6 expression was regulated by ERK1/2 signal pathway in MC3T3-E1 cells, but osteocalcin gene expression was not affected by ERK1/2 pathway. NF-kB signal pathway inhibitor PDTC significantly suppressed alkaline phosphatase activities in MC3T3-E1 cells under mechanical strain, and inhibited interleukin-6 gene expression. However, type I col agen and osteocalcin gene expression was not affected by NF-kB signal pathway. Results suggested that mechanical strain affected osteogenic differentiation and relevant gene expression in MC3T3-E1 cells by ERK1/2 and NF-kB signal pathway.