CAJ | 학술논문

[目的]探究促小鼠红细胞生成素（recombinant mouse Erythropoietin，rmEPO）对体外培养小鼠成骨细胞成骨活性的影响。[方法]3d内新生C57乳鼠10只，无菌条件下酶消化法分离其成骨细胞。原代培养成骨细胞传第一代，将生长良好的成骨细胞制成细胞悬液，随机分为空白组和rmEPO组（10ng·mL-1）。MTT比色法检测细胞增殖；比色法检测细胞碱性磷酸酶（alkaline phosphatase，ALP）活性；用茜素红染色观察细胞外基质矿化情况；用RT-PCR法检测细胞成骨相关基因（ALP、OPG、RANKL）的表达水平。[结果] rmEPO能够显著促进细胞增殖（与空白组相比P<0.05）、细胞ALP活性的表达（与空白组相比P<0.05）、细胞外基质矿化。能够显著上调ALP、OPG基因表达（与空白组相比P<0.05），下调RANKL基因的表达（与空白组相比P<0.05）。[结论]rmEPO能促进小鼠成骨细胞成骨活性。
[목적]탐구촉소서홍세포생성소（recombinant mouse Erythropoietin，rmEPO）대체외배양소서성골세포성골활성적영향。[방법]3d내신생C57유서10지，무균조건하매소화법분리기성골세포。원대배양성골세포전제일대，장생장량호적성골세포제성세포현액，수궤분위공백조화rmEPO조（10ng·mL-1）。MTT비색법검측세포증식；비색법검측세포감성린산매（alkaline phosphatase，ALP）활성；용천소홍염색관찰세포외기질광화정황；용RT-PCR법검측세포성골상관기인（ALP、OPG、RANKL）적표체수평。[결과] rmEPO능구현저촉진세포증식（여공백조상비P<0.05）、세포ALP활성적표체（여공백조상비P<0.05）、세포외기질광화。능구현저상조ALP、OPG기인표체（여공백조상비P<0.05），하조RANKL기인적표체（여공백조상비P<0.05）。[결론]rmEPO능촉진소서성골세포성골활성。
Objective] To investigate the role of rmEPO on cellosteogenic activity of osteoblast cells. [Methods] 10 C57 mice(3d old) were used, and the osteoblast cells were separated by enzyme digestion under the aseptic condition. Primary osteoblast cells were cultured to obtain the first generation of the cells, then divided into normal group and rmEPO group(10ng·min-1 rmEPO treated). We used the MTT colorimetric method to test cell proliferation assay, the colorimetric method to test the ALP activity, the alizarin red staining method to observe the calcified area, RT-PCR to test the expression of osteogenic genes(ALP,OPG,RANKL genes).[Results] After the rmEPO treatment, osteoblast cells showed increased cell viability compared with the normal group(P<0.05). And rmEPO-treated cells showed enhanced ALP activity and enhanced mineralization(P<0.05). rmEPO treatment also significantly promoted ALP and OPG expression(P<0.05) while significantly inhibiting RANKL expression(P<0.05). [Conclusion] rmEPO can induce the cellosteogenic activity of mouse osteoblast cells.