湖南中医药大学学报2025，Vol.45Issue(9)：1653-1662,10.DOI:10.3969/j.issn.1674-070X.2025.09.009
乳香-没药含药血清通过p38 MAPK信号通路促进BMMSCs成软骨分化的机制研究
Mechanism of Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum in promoting chondrogenic differentiation of BMMSCs through the p38 MAPK signaling pathway
摘要
Abstract
Objective To investigate the effects of Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum on chondrogenic differentiation of bone marrow mesenchymal stem cells(BMMSCs)and its mechanism of action.Methods The whole bone marrow adherence method was employed to isolate BMMSCs from the bilateral femur and tibia of rats,and the cell morphology was observed and identified by flow cytometry.The CCK-8 assay was used to determine the optimal concentration of Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum for promoting BMSC viability.Tumor necrosis factor-α(TNF-α)was used to induce BMMSCs to establish an in vitro inflammatory microenvironment,and its effective concentration was determined by CCK-8 assay.The BMMSCs were divided into blank group,TNF-α group,TNF-α+Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum group,and TNF-α+celecoxib group.Cells in each group were treated with 10 ng/mL TNF-α,10%Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum,or 10 μmol/L celecoxib.After the treatment,BMSC viability was measured by CCK-8 assay,apoptosis by TUNEL staining,glycosaminoglycan deposition in the extracellular matrix by Alcian blue staining,and expression of chondrogenic differentiation markers,including Sox9,collagenⅡ(ColⅡ),and aggrecan,as well as key proteins in the p38 mitogen-activated protein kinase(p38 MAPK)signaling pathway by Western blot.Results The isolated BMMSCs were spindle-or polygon-shaped,neatly arranged in a whirlpool-like pattern,positive for CD44,CD90,and CD105,and negative for CD34,CD45,and HLA-DR,consistent with the BMSC characteristics.The viability of BMMSCs treated with 2.5%,5%,and 10%Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum increased(P<0.05),while that treated with 20%and 25%Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum decreased(P<0.05);the BMSC viability treated with 5,10,15,20,25,and 30 ng/mL TNF-α was reduced(P<0.05).Compared with the blank group,the TNF-α group showed reduced viability(P<0.05),increased apoptosis(P<0.05),and fewer Alcian blue-stained cells.Additionally,the relative protein expression levels of Sox9,Col Ⅱ,and aggrecan were downregulated(P<0.05),while the ratios of p-p38/p38 and p-JNK/JNK were elevated(P<0.05).Compared with the TNF-α group,both the TNF-α+Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum and TNF-α+celecoxib groups showed improved viability(P<0.05),reduced apoptosis(P<0.05),and increased Alcian blue-stained cells.The relative protein expression levels of Sox9,Col Ⅱ,and aggrecan were upregulated(P<0.05),while the ratios of p-p38/p38 and p-JNK/JNK decreased(P<0.05).Conclusion Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum can promote the proliferation of BMMSCs in inflammatory environments and induce their chondrogenic differentiation.This mechanism may be related to the regulation of the p38 MAPK signaling pathway.关键词
乳香-没药含药血清/骨髓间充质干细胞/成软骨分化/肿瘤坏死因子-α/p38丝裂原活化蛋白激酶Key words
Ruxiang(Olibanum)and Moyao(Myrrha)-containing serum/bone marrow mesenchymal stem cells/chondrogenic differentiation/tumor necrosis factor-α/p38 mitogen-activated protein kinase分类
医药卫生引用本文复制引用
陆云,胡玉,陈军辉..乳香-没药含药血清通过p38 MAPK信号通路促进BMMSCs成软骨分化的机制研究[J].湖南中医药大学学报,2025,45(9):1653-1662,10.基金项目
新疆医科大学第六附属医院(第六临床医学院)科研专项基金项目(LFYKYZXJJ2024016) （第六临床医学院）
新疆医科大学第六附属医院临床药学重点专科基金资助项目(202401). （202401）
