湖南中医药大学学报2025，Vol.45Issue(4)：631-637,7.DOI:10.3969/j.issn.1674-070X.2025.04.007
地黄苷A对成骨细胞增殖分化及MCP-1/CCR2信号通路的影响
Effects of rehmannioside A on osteoblast proliferation,differentiation,and the MCP-1/CCR2 signaling pathway
摘要
Abstract
Objective To observe the effects of rehmannioside A on the osteoblast proliferation,differentiation,and its potential molecular mechanisms.Methods Osteoblast cell line MC3T3-E1 was used as an in vitro model.Cells were treated with different concentrations of rehmannioside A(0,1,10,100 μmol/L).The CCK-8 assay and bromodeoxyuridine(BrdU)fluorescence labeling method were employed to evaluate the effects of rehmannioside A on osteoblast proliferation at 24,48,and 72 h post-treatment.Experimental groups were divided as follows:blank group(saline),rehmannioside A group(10 μmol/L),and rehmannioside A+RS504393 group(10 μmol/L rehmannioside A+10 μg/mL RS504393).Alkaline phosphatase(ALP)activity of osteoblasts was determined using an ALP assay kit.RT-qPCR was performed to measure mRNA expression levels of osteogenic differentiation markers—osteocalcin(OCN),Runt-related transcription factor 2(Runx2),osterix(Osx),monocyte chemoattractant protein-1(MCP-1),and CC chemokine receptor-2(CCR2).Western blot analysis was used to measure protein expressions of OCN,Runx2,Osx,MCP-1,and CCR2.Results Compared with the 0 μmol/L and 1 μmol/L rehmannioside A groups,the 10 μmol/L rehmannioside A group showed increased OD values and BrdU fluorescence expressions at 48 and 72 h(P＜0.01),while the 100 μmol/L rehmannioside A group exhibited decreased OD values at 48 and 72 h(P＜0.05).Compared with the 10 μmol/L rehmannioside A group,the 100 μmol/L group had reduced OD values at 48 and 72 h(P＜0.05).Compared with the 0 μmol/L and 10 μmol/L rehmannioside A groups,the 100 μmol/L group showed decreased BrdU fluorescence expressions at 24,48,and 72 h(P＜0.01).Compared with the 1 μmol/L group,the 100 μmol/L group exhibited reduced BrdU fluorescence expressions at 48 and 72 h(P＜0.01).Compared with the blank group,the rehmannioside A group demonstrated increased ALP activity at 3,7,and 14 days(P＜0.01),with ALP activity progressively increasing over the prolonged culture time of osteoblasts(P＜0.01).However,compared with the rehmannioside A group,the rehmannioside A+RS504393 group showed decreased ALP activity at 3,7,and 14 days(P＜0.01),with ALP activity progressively increasing over the prolonged culture time of osteoblasts(P＜0.05).Compared with the blank group,the rehmannioside A group exhibited elevated mRNA and protein expression levels of OCN,Runx2,Osx,MCP-1,and CCR2(P＜0.01).Compared with the rehmannioside A group,the rehmannioside A+RS504393 group showed reduced mRNA and protein expressions of OCN,Runx2,Osx,MCP-1,and CCR2(P＜0.01).Conclusion Rehmannioside A can promote osteoblast proliferation and differentiation by activating the MCP-1/CCR2 signaling pathway.关键词
地黄苷A/成骨细胞/成骨分化/单核细胞趋化蛋白-1/类趋化因子受体-2Key words
rehmannioside A/osteoblasts/osteogenic differentiation/monocyte chemoattractant protein-1/CC chemokine receptor-2分类
中医学引用本文复制引用
刘杰,张慧霞,韩炜,谢荣,郑凯..地黄苷A对成骨细胞增殖分化及MCP-1/CCR2信号通路的影响[J].湖南中医药大学学报,2025,45(4):631-637,7.基金项目
新疆维吾尔自治区重点研发科研项目(2022XJKJ009). （2022XJKJ009）
