内源性透明质酸对小鼠牙乳头细胞增殖的影响

doi:10.13701/j.cnki.kqyxyj.2024.07.005

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (7): 593-598.DOI: 10.13701/j.cnki.kqyxyj.2024.07.005

内源性透明质酸对小鼠牙乳头细胞增殖的影响

黄海燕, 颜燕宏, 刘塬, 吴佳艳, 宋宸宇, 蒋备战^*

同济大学附属口腔医院儿童口腔科,同济大学口腔医学院,上海牙组织修复与再生工程技术研究中心,同济大学口腔医学研究所上海 200072

收稿日期:2024-01-10 出版日期:2024-07-28 发布日期:2024-07-24
通讯作者: *蒋备战,E-mail:jiangbeizhan@tongji.edu.cn
作者简介:黄海燕(1991~ ),女,安徽人,博士在读,研究方向:细胞外基质,牙发育。
基金资助:
国家自然科学基金(编号:8230035085);上海市卫生健康委员会科研项目(编号:202140357);上海申康医院发展中心新兴前沿技术联合攻关项目(编号:SHDC12023115)

Effect of Endogenous Hyaluronan on Proliferation of Mouse Dental Papilla Cells

HUANG Haiyan, YAN Yanhong, LIU Yuan, WU Jiayan, SONG Chenyu, JIANG Beizhan^*

Shanghai Engineering Research Center of Tooth Restoration and Regeneration ＆ Tongji Research Institute of Stomatology ＆ Department of Pediatric Dentistry, Stomatological Hospital and Dental School, Tongji University, Shanghai 200072, China.

Received:2024-01-10 Online:2024-07-28 Published:2024-07-24

摘要/Abstract

摘要： 目的:探究内源性透明质酸(hyaluronan,HA)对小鼠牙乳头细胞(mouse dental papilla cells,mDPCs)增殖的影响。方法:对出生后2.5 d的小鼠腹腔注射5-乙炔基-2′脱氧尿嘧啶核苷(5-ethynyl-2′-deoxyuridine, EdU)标记快速增殖细胞;体外分离培养mDPCs,使用不同浓度透明质酸酶(hyaluronidase,HAase)降解HA,细胞计数试剂盒(cell counting kit-8,CCK-8)法及EdU法检测细胞增殖;使用流式细胞术检测细胞周期分布和凋亡;转染siHyal2抑制HA降解,EdU法检测细胞增殖。结果:HA主要表达在小鼠切牙根尖慢增殖间充质细胞周围,而在转运扩增细胞(transit-amplifying cells,TACs)表达下降;CCK-8结果显示,相比对照组,800 μg/mL HAase明显促进细胞增殖(P＜0.0001);EdU检测及流式细胞术结果显示,HAase处理48 h后,细胞DNA合成增加,S期细胞比率明显增加(P＜0.01),并抑制早期细胞凋亡(P＜0.05);与之相反,当HA积聚在mDPCs细胞周,细胞增殖受到抑制(P＜0.01)。结论:内源性HA与mDPCs增殖密切相关,HA分解可促进细胞增殖;而局部HA的积聚则抑制细胞增殖。

关键词: 细胞外基质, 透明质酸, 透明质酸酶2, 细胞增殖

Abstract: Objective: To explore the effect of endogenous hyaluronan (HA) remodeling on the proliferation of mouse dental papilla cells (mDPCs). Methods: Fast-proliferating cells were labeled with EdU by intraperitoneal injection into mice at postnatal day 2.5. mDPCs were isolated, and then treated with different concentrations of hyaluronidase (HAase). The cell proliferation was detected by CCK-8 assay and EdU assay. Flow cytometry was used to detect cell cycle distribution and apoptosis. The transfection of siHyal2 caused HA accumulation and the cell proliferation was detected by EdU assay. Results: HA expression was mainly observed in the slow-proliferating mesenchymal cells in mouse incisors, while the expression of HA was decreased in transit-amplifying cells (TACs). The CCK-8 results indicated that 800 μg/mL HAase significantly promoted cell proliferation compared with the control group (P＜0.0001). After 48 h of HAase treatment, the EdU assay and flow cytometry results showed an increase in cellular DNA synthesis and a significant increase in the ratio of S-phase cells (P＜0.01). Additionally, early cell apoptosis was inhibited (P＜0.05). In contrast, the accumulation of HA around mDPCs was found to significantly inhibit cell proliferation (P＜0.01). Conclusion: The proliferation of mDPCs is closely related to endogenous HA. The degradation of HA promotes cell proliferation, which is inhibited by local HA accumulation.

Key words: extracellular matrix, hyaluronan, hyaluronidase 2, cell proliferation

黄海燕, 颜燕宏, 刘塬, 吴佳艳, 宋宸宇, 蒋备战. 内源性透明质酸对小鼠牙乳头细胞增殖的影响[J]. 口腔医学研究, 2024, 40(7): 593-598.

HUANG Haiyan, YAN Yanhong, LIU Yuan, WU Jiayan, SONG Chenyu, JIANG Beizhan. Effect of Endogenous Hyaluronan on Proliferation of Mouse Dental Papilla Cells[J]. Journal of Oral Science Research, 2024, 40(7): 593-598.

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内源性透明质酸对小鼠牙乳头细胞增殖的影响

Effect of Endogenous Hyaluronan on Proliferation of Mouse Dental Papilla Cells

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