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

作　　者：黄海燕 颜燕宏 刘塬 吴佳艳 宋宸宇 蒋备战[1] 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)

机构地区：[1]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

出　　处：《口腔医学研究》2024年第7期593-598,共6页Journal of Oral Science Research

基　　金：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)。

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,800μg/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。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 si Hyal2 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 800μg/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.

关 键 词：细胞外基质 透明质酸 透明质酸酶2 细胞增殖 

分 类 号：R78[医药卫生—口腔医学]