黄芪多糖对高糖环境下MC3T3-E1细胞活性影响的研究

Effect of astragalus polysaccharides on the activities of osteoblast in high glucose conditions

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/m L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、OCNm RNA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 m RNA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。

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.