柚皮素通过基质细胞衍生因子1/趋化因子受体4信号轴对脂多糖作用下人牙周膜干细胞抗炎、成血管和成骨分化能力的影响

doi:10.7518/hxkq.2023.2022293

华西口腔医学杂志 ›› 2023, Vol. 41 ›› Issue (2): 175-184.doi: 10.7518/hxkq.2023.2022293

柚皮素通过基质细胞衍生因子1/趋化因子受体4信号轴对脂多糖作用下人牙周膜干细胞抗炎、成血管和成骨分化能力的影响

李胜鸿¹^,²(), 彭世元¹^,², 罗小玲³, 王奕佩⁴, 徐晓梅¹^,²()

^1.西南医科大学附属口腔医院正畸科，泸州 646000
^2.口颌面修复重建和再生泸州市重点实验室，泸州 646000
^3.简阳市人民医院口腔科，简阳 641400
^4.自贡市第一人民医院口腔科，自贡 643000

收稿日期:2022-07-26 修回日期:2022-12-27 出版日期:2023-04-01 发布日期:2023-04-14
通讯作者: 徐晓梅 E-mail:361951960@qq.com;xuxiaomei@swmu.edu.cn
作者简介:李胜鸿，医师，硕士，E-mail：361951960@qq.com
基金资助:
四川省科技厅应用基础项目(2021YJ0151);泸州市科技局重点项目(2020LZXNYDZ06)

Effect of naringenin on the anti-inflammatory, vascularization, and osteogenesis differentiation of human periodontal ligament stem cells via the stromal cell-derived factor 1/C-X-C motif chemokine receptor 4 signaling axis stimulated by lipopolysaccharide

Li Shenghong¹^,²(), Peng Shiyuan¹^,², Luo Xiaoling³, Wang Yipei⁴, Xu Xiaomei¹^,²()

^1.Dept. of Orthodontics, The Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, China
^2.Oral＆Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, Luzhou 646000, China
^3.Dept. of Stomatology, The People’s Hospital of Jianyang City, Jianyang 641400, China
^4.Dept. of Stomatology, Zigong First People’s Hospital, Zigong 643000, China

Received:2022-07-26 Revised:2022-12-27 Online:2023-04-01 Published:2023-04-14
Contact: Xu Xiaomei E-mail:361951960@qq.com;xuxiaomei@swmu.edu.cn
Supported by:
Application Foundation Project of Science and Technology Department of Sichuan Province(2021YJ-0151);Key Project of Luzhou Science and Technology Bureau(2020LZXNYDZ06);Correspondence: Xu Xiaomei, E-mail: xuxiaomei@swmu.edu.cn

摘要/Abstract

摘要：

目的探究柚皮素（Nar）对脂多糖（LPS）刺激下的人牙周膜干细胞（hPDLSCs）抗炎、成血管和成骨能力的影响及其机制。方法采用细胞计数试剂盒（CCK-8）、划痕试验和Transwell实验研究hPDLSCs的增殖和迁移能力。采用碱性磷酸酶（ALP）染色、茜素红染色、管腔形成实验、酶联免疫吸附实验（ELISA）、实时荧光定量逆转录聚合酶链反应（qRT-PCR）和蛋白印迹实验（Western blot）检测hPDLSCs抗炎、成血管和成骨分化能力。结果 10 μmol/L Nar可减轻10 μg/mL LPS刺激的hPDLSCs炎症反应，促进其增殖、迁移和成血管分化，0.1 μmol/L Nar可有效恢复10 μg/mL LPS刺激的hPDLSCs成骨能力。加入CXCR4抑制剂AMD3100后，Nar促进抗炎和成骨分化的作用降低，炎性hPDLSCs成血管分化升高。结论 Nar促进了hPDLSCs抗炎、成血管和成骨分化，该作用与基质细胞衍生因子1/趋化因子受体4信号轴有关。

关键词: 柚皮素, 炎性人牙周膜干细胞, 抗炎, 成骨, 成血管

Abstract:

Objective This study aimed to investigate how naringenin (Nar) affected the anti-inflammatory, vascula-rization, and osteogenesis differentiation of human periodontal ligament stem cells (hPDLSCs) stimulated by lipopolysaccharide (LPS) and to preliminarily explore the underlying mechanism. Methods Cell-counting kit-8 (CCK8), cell scratch test, and Transwell assay were used to investigate the proliferation and migratory capabilities of hPDLSCs. Alkaline phosphatase (ALP) staining, alizarin red staining, lumen-formation assay, enzyme-linked immunosorbent assay, quantitative timed polymerase chain reaction, and Western blot were used to measure the expression of osteopontin (OPN), Runt-related transcription factor 2 (RUNX2), vascular endothlial growth factor (VEGF), basic fibroblast growth factor (bFGF), von Willebrand factor (vWF), tumor necrosis factor-α (TNF-α), and interleukin (IL)-6. Results We observed that 10 μmol/L Nar could attenuate the inflammatory response of hPDLSCs stimulated by 10 μg/mL LPS and promoted their proliferation, migration, and vascularization differentiation. Furthermore, 0.1 μmol/L Nar could effectively restore the osteogenic differentiation of inflammatory hPDLSCs. The effects of Nar’s anti-inflammatory and promotion of osteogenic differentiation significantly decreased and inflammatory vascularization differentiation increased after adding AMD3100 (a specific CXCR4 inhibitor). Conclusion Nar demonstrated the ability to promote the anti-inflammatory, vascularization, and osteogenic effects of hPDLSCs stimulated by LPS, and the ability was associated with the stromal cell-derived factor/C-X-C motif chemokine receptor 4 signaling axis.

Key words: naringenin, inflammatory human periodontal ligament stem cells, anti-inflammatory, osteogenesis, vascularization

中图分类号:

Q 257

李胜鸿, 彭世元, 罗小玲, 王奕佩, 徐晓梅. 柚皮素通过基质细胞衍生因子1/趋化因子受体4信号轴对脂多糖作用下人牙周膜干细胞抗炎、成血管和成骨分化能力的影响[J]. 华西口腔医学杂志, 2023, 41(2): 175-184.

Li Shenghong, Peng Shiyuan, Luo Xiaoling, Wang Yipei, Xu Xiaomei. Effect of naringenin on the anti-inflammatory, vascularization, and osteogenesis differentiation of human periodontal ligament stem cells via the stromal cell-derived factor 1/C-X-C motif chemokine receptor 4 signaling axis stimulated by lipopolysaccharide[J]. West China Journal of Stomatology, 2023, 41(2): 175-184.

图/表 5

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参考文献 33

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基因名称	引物序列（5’-3’）
RUNX2	Forward：GAACCCAGAAGGCACAGACA Reverse：GGCTCAGGTAGGAGGGGTAA
OPN	Forward：CACCGAGACACCATGAGAGC Reverse：CTGCTTGGACACAAAGGCTGC
bFGF	Forward：GCGACCCTCACATCAAGCTA Reverse：AGCCAGGTAACGGTTAGCAC
VEGF	Forward：ATCGAGTACATCTTCAAGCCAT Reverse：GTGAGGTTTGATCCGCATAATC
vWF	Forward：TTGACGGGGAGGTGAATGTG Reverse：ATGTCTGCTTCAGGACCACG
SDF-1	Forward：TGCCCTTCAGATTGTAGCCC Reverse：GTGGGTCTAGCGGAAAGTCC
CXCR4	Forward：CGTGAATGAGTGTCTAGGCAGG Reverse：GGCTTTGGTTTTAAGTGCCATC
TNF-α	Forward：AGCTGGTGGTGCCATCAGAGG Reverse：TGGTAGGAGACGGCGATGCG
IL-6	Forward：CACTGGTCTTTTGGAGTTTGAG Reverse：GGACTTTTGTACTCATCTGCAC
β-actin	Forward：CGACCCGTCTACAGGTGAAC Reverse：GAGAGGGACTCAATCAGCCG

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柚皮素通过基质细胞衍生因子1/趋化因子受体4信号轴对脂多糖作用下人牙周膜干细胞抗炎、成血管和成骨分化能力的影响

Effect of naringenin on the anti-inflammatory, vascularization, and osteogenesis differentiation of human periodontal ligament stem cells via the stromal cell-derived factor 1/C-X-C motif chemokine receptor 4 signaling axis stimulated by lipopolysaccharide

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