口腔疾病防治2018，Vol.26Issue(3)：158-165,8.DOI:10.12016/j.issn.2096-1456.2018.03.004
银杏叶中异鼠李素对RAW264.7细胞向破骨细胞分化的影响及其分子机制
Effect and molecular mechanism of isorhamnetin extracted from Ginkgo biloba on the differentiation of RAW264.7 cells into osteoclasts
摘要
Abstract
Objective To investigate the effect and potential molecular mechanisms of isorhamnetin (ISO) extracted from Ginkgo biloba on the differentiation of osteoclasts. Methods Osteoclast precursor RAW264.7 cells were induced with RANKL to differentiate into mature osteoclasts. Different concentrations of ISO were added to RAW 264.7 cells to determine its effect on osteoclast differentiation. CCK8 was used to evaluate the effect of ISO on cytotoxicity. The im-pact of ISO on the osteoclast differentiation process was investigated by analyzing tartrate resistance and bone resorption lacuna. Real-time PCR was performed to analyze the levels of differentiation marker genes, including tartrate resistant acid phosphatase (Trap), cathepsin K (Ctsk), and matrix metalloproteinase 9 (MMP-9);differentiation-related transcription factors, including the proto-oncogene protein c-Fos, nuclear factor of activated T-cells cytoplasmic 1(NFATc1);and the levels of downstream NF-κB p65 signaling pathway phosphorylation. Using the above-described method, we verified that ISO exerted an inhibitory effect on osteoclast differentiation and explored related molecular mechanisms. Results Dif-ferent concentrations of ISO (1-10 μM) had no cytotoxic effects on RAW264.7 cells, inhibited TRAP activity and decreased the number of bone resorption lacuna during osteoclast differentiation. When applied at a concentration of 10 μM, its inhibitory effect was significant. In addition, ISO significantly reduced the expression levels of Trap, Ctsk, MMP-9, c-Fos, NFATc1 and NF-κB p65 mRNA. Conclusion ISO extracted from Ginkgo biloba extract exerted an inhibitory effect on osteoclast differentiation, and the mechanism underlying its activity may involve the inhibition of the classical NF-κB pathway.关键词
银杏叶提取物/黄酮类化合物/异鼠李素/破骨细胞/破骨分化/核因子κB受体活化因子配基/核因子κB受体活化因子Key words
Ginkgo biloba extract/Flavonoids/Isorhamnetin/Osteoclast/Osteoclast differentiation/RANKL/NF-κB分类
医药卫生引用本文复制引用
李婧,程梁,郭吕华,李彤,杨沫扬,王俊妹,吴哲..银杏叶中异鼠李素对RAW264.7细胞向破骨细胞分化的影响及其分子机制[J].口腔疾病防治,2018,26(3):158-165,8.基金项目
吉林省教育厅"十二五"科学技术研究项目(吉教科合字[2015]第530号) （吉教科合字[2015]第530号）
广东省科技计划国际合作项目(2017A050501041) （2017A050501041）
