摘要
目的:研究槲皮素(QUE)对骨髓间充质干细胞(BMSCs)成骨及成脂分化的影响,及雌激素受体(ER)信号通路对其的调控作用。方法:全骨髓贴壁法提取大鼠BMSCs;细胞形态学及表面抗原鉴定BMSCs;CCK8法测定QUE对BMSCs活力的影响;碱性磷酸酶(ALP)法及ELISA法分别检测QUE干预BMSCs后细胞培养上清液中ALP和BGP的表达;加入ER阻断剂ICI182780,Western blot及实时荧光定量PCR法检测正常及ER阻断后QUE对BMSCs成脂及成骨分化相关因子表达的影响。结果:细胞呈长梭形、多角状,表面抗原CD29阳性率99.93%,CD45阳性率3.20%;综合不同浓度QUE对BMSCs增殖和成骨分化影响,选取10^(-5)mol/L QUE进行后续实验。QUE能促进RUNX2及ALP的表达,抑制PPARγ及C/EBPα的表达(P<0.05);与QUE组比较,QUE与ICI182780联用后可显著抑制ALP的表达,促进PPARγ的表达(P<0.05),但RUNX2与C/EBPα的蛋白及mRNA变化不一致。结论:QUE促进BMSCs成骨分化、抑制成脂分化,可能是通过ER信号通路介导完成。
Objective: To investigate the effects of quercetin on osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs) and its relationship with estrogen signaling pathway. Methods: Whole bone marrow adherent method was used to extract the BMSCs, cell morphological and surface antigen were used to identify the BMSCs, CCK8 assay was used to determinate the cell vitality under the treatment of quercetin, the alkaline phosphatase kit and ELISA kits used to evaluate the osteogenesis ability of BMSCs, Western blot and qPCR used to detect the osteogenic and adipogenic differentiation. Results: The morphology of cells was fusiform and polygonal, 99.93% CD29 was positively expressed, while 3.2% CD45 was positively expressed;integrating the effects of different concentrations of QUE on BMSCs proliferation and osteogenic differentiation, 10^(-5) mol/L QUE was selected as the intervention concentration at the following experiments. Quercetin promoted the expression of RUNX2 and ALP, and inhibited the expression of PPARγ and C/EBPα(P<0.05). Compared with the QUE group, the expression of ALP significantly attenuated, while PPARγ obviously increased(P<0.05), but the changes of protein and mRNA of RUNX2 and C/EBPα were not consistent. Conclusion: Quercetin at a certain concentration could promote the proliferation and osteogenic differentiation of BMSCs, which effects may be mediated by estrogen receptor signaling.
作者
李小云
杨丽
张荣华
LI Xiao-yun;YANG Li;ZHANG Rong-hua(Jinan University,Guangzhou 510632,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2020年第12期6011-6014,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81873202)
国家重点研发计划项目(No.2018YFC2002500)。
关键词
槲皮素
雌激素受体信号通路
骨髓间充质干细胞
成骨分化
成脂分化
Quercetin
Estrogen receptor signaling
Bone mesenchymal stem cell
Osteogenic differentiation
Adipogenic differentiation
