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研究强磁重力环境(high magneto-gravitational environment,HMGE)对不同分化阶段MC3T3-E1成骨细胞分化的影响及其可能机制。采用HMGE的μg[9 Tesla(T)]、1 g(16 T)和2 g(12 T)以及正常对照(1 g,地磁场)环境,分别对诱导成骨分化4 d和7 d的MC3T3-E1细胞处理12 h,对硝基苯磷酸法检测成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)活性;茜素红S染色检测细胞矿化结节形成情况;Real-time RT-PCR检测成骨分化相关分子ALP、runt-related transcription factor 2(Runx2),I型胶原α1(type I collagenα1,Col Iα1)、骨钙素(osteocalcin,OC)以及牙本质基质蛋白1(dentin matrixprotein 1,DMP1)mRNA的表达。结果发现,经过12 h的HMGE处理后,与对照组(C)相比,μg组和1 g组成骨分化7 d的MC3T3-E1细胞ALP活性显著增加(P<0.001,P<0.01),而HMGE对成骨分化4 d细胞的ALP活性无显著影响。茜素红S染色显示,μg组和1 g组成骨分化7 d的MC3T3-E1细胞形成矿化结节较对照组增加。Real-time RT-PCR检测显示,HMGE处理12 h后,成骨分化4 d与7 d细胞成骨分化相关基因(ALP、Runx2、Col Iα1、OC和DMP1)在μg组和1 g组的表达明显上调,而2 g组下调(P<0.05,P<0.01,P<0.001)。结果表明,处于4 d与7 d不同分化阶段的MC3T3-E1细胞对12 h HMGE环境的处理敏感程度不同,7 d矿化阶段的细胞对HMGE更敏感,同时,磁场表现出促进成骨细胞分化的作用。
To investigate the effect of high magneto-gravitational environment (HMGE) on the differentiation of MC3T3-E1 osteoblasts and its possible mechanism. The MC3T3 cells induced by osteogenic differentiation for 4 d and 7 d were treated with HMGE of μg [9 Tesla (T)], 1 g (16 T) and 2 g (12 T) and normal control (1 g, -E1 cells for 12 h, alkaline phosphatase (ALP) activity of osteoblasts was detected by p-nitrophenyl phosphate assay; alizarin red S staining was used to detect the formation of mineralized nodules; Real-time RT-PCR The expression of ALP, runt-related transcription factor 2 (Runx2), type I collagenα1 (Col Iα1), osteocalcin (OC) and dentin matrix protein 1 ) mRNA expression. The results showed that the ALP activity of MC3T3-E1 cells treated with 1 μg of osteogenic differentiation group increased significantly (P <0.001, P <0.01) compared with control group (C) after 12 h of HMGE treatment HMGE had no significant effect on the ALP activity of osteogenic differentiation 4 d cells. Alizarin red S staining showed that the formation of mineralized nodules of MC3T3-E1 cells in the μg group and 1 g of osteogenic differentiation for 7 days increased compared with the control group. The expression of ALP, Runx2, Col Iα1, OC and DMP1 in the groups of μg and 1 g at 4 d and 7 d after osteogenic differentiation was detected by Real-time RT-PCR (P <0.05, P <0.01, P <0.001). The results showed that MC3T3-E1 cells at 4 and 7 d differentiation stages were sensitive to 12 h HMGE treatment, and the cells at 7 d mineralization stage were more sensitive to HMGE. Meanwhile, the magnetic field showed osteoblast differentiation Role.