摘要
目的探究莫诺苷在体外对MC3T3-E1成骨细胞增殖、分化、矿化、游离钙的影响。方法分别用MTT比色法,碱性磷酸酶(ALP)试剂盒,骨钙素(BGP)试剂盒,透射电镜进行观察,采用Flou-3的负载检测不同浓度莫诺苷对成骨细胞增殖、分化、矿化、细胞内游离钙的影响。结果MTT比色法结果表明,2.5、12.5、62.5、125 mg·L^(-1)的莫诺苷对MC3T3-E1细胞增殖作用不显著(P>0.05);与空白组比,2.5、12.5 mg·L^(-1)的莫诺苷可明显提高MC3T3-E1细胞ALP活性,而62.5 mg·L^(-1)的莫诺苷仅在作用2 d时明显提高ALP活性(P<0.05);2.5 mg·L^(-1)的莫诺苷用药5 d可显著提高MC3T3-E1细胞BGP活性(P<0.05);矿化结果显示培养6 d的给药组和空白组都没有出现直径大于200μm的矿化结节,培养12 d的给药组、空白组均出现直径大于200μm的橘红色矿化结节,给药组矿化结节数高于空白组(P<0.05);电镜观察结果显示用药组细胞外可见密度较大的矿化物质;Flou-3的负载实验结果显示2.5 mg·L^(-1)的莫诺苷组使成骨细胞内平均荧光强度明显升高(P<0.05)。结论莫诺苷能够促进MC3T3-E1成骨细胞分化、矿化以及内钙释放。
Objective To determine the effect of monosine on the proliferation,differentiation,mineralization and free calcium of MC3T3-E1 osteoblasts in vitro.Methods MTT colorimetric assay,alkaline phosphatase(ALP)kit,osteocalcin(BGP)kit,transmission electron microscopy,and Flou-3 loading were used to observe the effect of different concentrations of monosine on the proliferation,differentiation,mineralization,and intracellular free calcium of osteoblasts.Results MTT colorimetric assay showed monosine at 2.5,12.5,62.5,and 125 mg·L^(-1)had no significant effect on the proliferation of MC3T3-E1 cells(P>0.05).Compared with the blank group,monosine at 2.5 and 12.5 mg·L^(-1)obviously increased the ALP activity of MC3T3-E1 cells,monosine at 62.5 mg·L^(-1)increased the ALP activity only after 2 days of action(P<0.05),monosine at 2.5 mg·L^(-1)greatly increased the BGP activity of MC3T3-E1 cells after 5 days of cell administration(P<0.05).The mineralization showed no mineralized nodules with diameter over 200μm in both the administration group and the blank group after 6 days of culture.While orange mineralized nodules were found with diameter over 200μm in both the administration group and the blank group after 12 days of culture,statistically different from the blank group(P<0.05).The electron microscopy showed bigger extracellular density of mineralized substances in the drug group.The Flou-3 loading experiment showed that the average fluorescence intensity of osteoblasts in the 2.5 mg·L^(-1)monosine group was significantly higher than that in the blank group(P<0.05).Conclusion Monosine can promote the differentiation,mineralization and calcium release of MC3T3-E1 osteoblasts.
作者
袁旭
董培良
韩华
YUAN Xu;DONG Pei-liang;HAN Hua(Institute of Traditional Chinese Medicine,Heilongjiang University of Chinese Medicine,Harbin 150000;College of Pharmacy,Heilongjiang University of Traditional Chinese Medicine,Harbin 150000)
出处
《中南药学》
CAS
2022年第12期2799-2804,共6页
Central South Pharmacy
基金
黑龙江省普通高等学校青年学术骨干支持计划项目(No.1153G038)
黑龙江中医药大学博士创新基金项目(No.B201003)。
