牵张应力介导细胞成骨分化及相关基因的表达  

Osteogenic differentiation and related gene expression mediated by mechanical strain

作　　者：刘名燕[1] 李燕[1] 钱红[2] 冯云霞[1] 段银钟[2] 李永明[2] 

机构地区：[1]山西医科大学口腔系,山西省太原市030001 [2]解放军第四军医大学口腔医学院正畸科,陕西省西安市710032

出　　处：《中国组织工程研究》2013年第50期8629-8634,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30970697;31370943);山西医科大学博士启动基金项目(B03201219)~~

摘　　要：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。BACKGROUND： The regulatory role of extracellular signal regulated kinase 1/2 （ERK1/2） and nuclear factor kappa B （NF-KB） signal pathways in the osteogenic differentiation of MC3T3-E1 cells subjected to mechanical strain remains unclear. OBJECTIVE： To investigate the effects of ERKI/2 and NF-kB signal pathway on alkaline phosphatase, type 1 collagen, 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 ERKI/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 I collagen, osteocalcin and interleukin-6 mRNA expression before and after cell loading. RESULTS AND CONCLUSION： Under 12% mechanical strain, alkaline phosphatase, type I collagen, and interleukin-6 expression was regulated by ERKI/2 signal pathway in MC3T3-E1 cells, but osteocalc!n gene expression was not affected by ERKI/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 collagen and osteecalcin 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 ERKI/2 and NF-kB signal pathway.

关 键 词：组织构建 骨组织构建 牵张应力 成骨细胞 ERK1 2 核因子KB 信号转导 碱性磷酸酶 I型胶原 白细胞介素6 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]