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西红花苷通过Wnt/β-连环蛋白信号通路对人牙周膜干细胞的成骨作用

Osteogenic effect of crocin in human periodontal ligament stem cells via Wnt/β-catenin signaling.

作者信息

Gu Yingzhi, Bai Yuxing

机构信息

Department of Orthodontics, Beijing Stomatological Hospital, Capital Medical University, Beijing, China.

出版信息

Oral Dis. 2024 Apr;30(3):1429-1438. doi: 10.1111/odi.14523. Epub 2023 Feb 13.

Abstract

OBJECTIVES

Crocin is a major class of medicinal components in saffron. This study aimed to determine whether crocin directly promotes the proliferation and osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) in vitro and in vivo.

MATERIALS AND METHODS

CCK8 cell proliferation assay, reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blot analysis and Alizarin Red staining were performed in PDLSCs using crocin as a stimulant. DKK1 was used to selectively inhibit Wnt/β-catenin signaling, and Western blotting was performed to investigate the underlying mechanism. The PDLSCs were mixed with calcium phosphate cement and implanted into nude mice subcutaneously to study the effect of crocin on PDLSC osteogenic differentiation in vivo.

RESULTS

The CCK-8 assay showed that crocin did not promote the proliferation of PDLSCs. Treatment with 400 μM crocin significantly promoted PDLSC mRNA levels of ALP, COL1 and OCN; RUNX2 and BMP2 protein expression; mineralized nodule formation in vitro and in vivo; and ALP activity in tissues in vivo. In addition, crocin significantly promoted the phosphorylation of β-catenin and cyclin D1. DKK1 inhibits Wnt/β-catenin activation and partially reverses crocin-mediated promotion of PDLSC osteogenic differentiation.

CONCLUSION

Crocin may contribute to the regeneration of periodontal bone tissue.

摘要

目的

西红花苷是藏红花中的一类主要药用成分。本研究旨在确定西红花苷在体外和体内是否能直接促进人牙周膜干细胞(PDLSCs)的增殖和成骨分化。

材料与方法

以西红花苷为刺激物,对PDLSCs进行CCK8细胞增殖试验、逆转录定量聚合酶链反应(RT-qPCR)、蛋白质免疫印迹分析和茜素红染色。使用DKK1选择性抑制Wnt/β-连环蛋白信号通路,并通过蛋白质免疫印迹法研究其潜在机制。将PDLSCs与磷酸钙骨水泥混合后皮下植入裸鼠体内,以研究西红花苷对PDLSCs体内成骨分化的影响。

结果

CCK-8试验表明,西红花苷不促进PDLSCs的增殖。400μM西红花苷处理显著促进了PDLSCs中碱性磷酸酶(ALP)、I型胶原蛋白(COL1)和骨钙素(OCN)的mRNA水平;RUNX2和骨形态发生蛋白2(BMP2)的蛋白表达;体内外矿化结节的形成;以及体内组织中的ALP活性。此外,西红花苷显著促进了β-连环蛋白和细胞周期蛋白D1的磷酸化。DKK1抑制Wnt/β-连环蛋白激活,并部分逆转了西红花苷介导的PDLSCs成骨分化促进作用。

结论

西红花苷可能有助于牙周骨组织的再生。

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