CAJ | 학술논문

目的：：探讨高糖环境下黄芪多糖(伯恩)对细胞MC3T3-E1增殖、分化和矿化的影响。方法：将MC3T3-E1分别与含黄芪多糖浓度为5、0.5、0.05 mg/mL的高糖培养基(分别为HDA组、MDA组、LDA组)和单纯高糖培养基(对照组)共同培养后,分别通过MTT法、碱性磷酸酶和茜素红染色法观察各组MC3T3-E1细胞的增殖、分化和矿化活性；激光共聚焦显微镜观察各组细胞的超微骨架结构；实时荧光定量PCR检测各组细胞中骨活性相关基因 Runx-2、OPN、OCNmRNA 的表达水平。结果：在高糖环境下 LDA、MDA 组更利于MC3T3-E1细胞的增殖和分化,与HDA组和对照组相比P<0.05； LDA组MC3T3-E1细胞的矿化程度及细胞超微骨架结构均明显优于其他各组(P<0.05)； RT-PCR检测显示,LDA组能显著增加 MC3T3-E1细胞中OPN、Runx-2、OCN mRNA的表达水平(P<0.05)。结论：高糖环境下,LDA组有利于MC3T3-E1细胞的增殖、分化和矿化；其对细胞活性的影响,与促进细胞骨架的排列、伸展和增加Runx-2、OPN、OCN的表达量有关。
목적：：탐토고당배경하황기다당(백은)대세포MC3T3-E1증식、분화화광화적영향。방법：장MC3T3-E1분별여함황기다당농도위5、0.5、0.05 mg/mL적고당배양기(분별위HDA조、MDA조、LDA조)화단순고당배양기(대조조)공동배양후,분별통과MTT법、감성린산매화천소홍염색법관찰각조MC3T3-E1세포적증식、분화화광화활성；격광공취초현미경관찰각조세포적초미골가결구；실시형광정량PCR검측각조세포중골활성상관기인 Runx-2、OPN、OCNmRNA 적표체수평。결과：재고당배경하 LDA、MDA 조경리우MC3T3-E1세포적증식화분화,여HDA조화대조조상비P<0.05； LDA조MC3T3-E1세포적광화정도급세포초미골가결구균명현우우기타각조(P<0.05)； RT-PCR검측현시,LDA조능현저증가 MC3T3-E1세포중OPN、Runx-2、OCN mRNA적표체수평(P<0.05)。결론：고당배경하,LDA조유리우MC3T3-E1세포적증식、분화화광화；기대세포활성적영향,여촉진세포골가적배렬、신전화증가Runx-2、OPN、OCN적표체량유관。
AIM:To evaluate the effect of Astragalus polysaccharides ( APS) on the proliferation, differentia-tion and mineralization of MC3T3-E1 cells in high glucose conditions and to study the underlying mechanisms. METH-ODS:MC3T3-E1 cells were cultured in high-glucose medium containing 0. 05(LDA), 0. 5(MDA) and 5(HDA) mg/mL of APS respectively. Cell proliferation, differentiation and mineralization were examined by MTT assay, ALP Kit and Alizarin red dye staining respectively. The expression of Runx-2, OPN and OCN were examined by RT-PCR. The cytoskeleton of the cells was observed by confocal laser scanning microscopy ( CLSM) . RESULTS: APS promoted the proliferation of MC3T3-E1 cells by LDA and MDA(P<0. 05). LDA and MDA significantly increased the ALP activity and calcified nodules (P<0. 05). Furthermore, LDA increased the gene expressions of Runx-2, OPN and OCN in a time-dependent manner (P<0. 05). CLSM observation showed that the cells in LDA group were more regular in struc-ture and had more extensions than those in the other groups at each time point. CONCLUSION: Under high glucose conditions, 0. 05 mg/mL APS promotes the proliferation, differentiation and mineralization of MC3T3-E1 cells. The effect is related to its promotion of the expression of OPN, Runx-2 and OCN.