重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响

doi:10.19439/j.sjos.2020.01.002

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (1): 7-12.doi: 10.19439/j.sjos.2020.01.002

重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响

曹颖, 彭伟伟, 王璇, 伍甜甜, 杜嵘^*, 朱亚琴^*

上海交通大学医学院附属第九人民医院·口腔医学院口腔综合科,国家口腔疾病临床医学研究中心, 上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011

收稿日期:2018-12-20 出版日期:2020-02-25 发布日期:2020-03-09
通讯作者: 朱亚琴,E-mail：zyq1590@163.com;杜嵘,E-mail：summerdr@163.com。^*共同通信作者
作者简介:曹颖（1993-）,女,硕士研究生,E-mail：caoy_0425@126.com
基金资助:
国家自然科学基金（81271134、81700949）

Effect of recombinant connective tissue growth factor on proliferation and odontogenic differentiation of dental pulp cells

CAO Ying, PENG Wei-wei, WANG Xuan, WU Tian-tian, DU Rong, ZHU Ya-qin

Department of General Dentistry, Shanghai Ninth People's Hospital,College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology. Shanghai 200011, China

Received:2018-12-20 Online:2020-02-25 Published:2020-03-09

摘要/Abstract

摘要： 目的：研究重组人结缔组织生长因子（recombinant connective tissue growth factor, rCTGF）对牙髓细胞（human dental pulp cells,hDPCs）增殖及分化的影响。方法：利用不同浓度（0、1、10、100 ng/mL）rCTGF分别处理牙髓细胞,CCK8法检测牙髓细胞增殖情况;茜素红染色和半定量试验检测细胞矿化结节的形成变化,qRT-PCR测定成牙本质分化相关基因DMP-1、DSPP和OC的表达情况,Western 免疫印迹法测定rCTGF刺激牙髓细胞后,ERK1/2信号通路的磷酸化水平。采用SAS 9.3软件包对数据进行统计学分析。结果：高浓度的rCTGF（100 ng/mL）可以促进牙髓细胞增殖;经矿化诱导后,10 ng/mL rCTGF促进牙髓细胞矿化结节形成的效果最好,钙盐沉积量最明显（P<0.05）,成牙本质分化相关基因DMP-1、DSPP的表达显著上调（P<0.05）。Western 免疫印迹结果显示,10 ng/mL rCTGF刺激牙髓细胞后,p-ERK1/2蛋白的表达升高。结论：rCTGF可能通过激活ERK1/2信号通路,促进牙髓细胞的增殖与分化。

关键词: 重组人结缔组织生长因子, 牙髓细胞, 细胞增殖, 细胞分化, ERK1/2信号通路

Abstract: PURPOSE: To analyze the effect of recombinant connective tissue growth factor(rCTGF) on proliferation and differentiation of human dental pulp cells(hDPCs). METHODS: Human dental pulp cells were cultured in vitro and treated with rCTGF at different concentrations (0, 1, 10, 100 ng/mL). The proliferation of dental pulp cells was detected by CCK8 assay. The formation of mineralized nodules was determined by alizarin red staining and half-quantitative alizarin Red S assay. qRT-PCR was utilized to detect the expression of odontogenic differentiation related genes DMP-1, DSPP and OC, and the phosphorylation level of ERK1/2 signaling pathway was detected by Western blot. The data were analyzed with SAS 9.3 software package. RESULTS: High concentration of rCTGF(100 ng/mL) could promote proliferation of dental pulp cells. After mineralization induction, 10 g/mL rCTGF had the best effect on promoting the formation of mineralized nodules in dental pulp cells, and calcium ion deposition was the most obvious(P<; 0.05). The expression of odontogenic differentiation related genes DMP-1 and DSPP was significantly up-regulated(P<; 0.05). Western blot results showed that hDPCs stimulated by 10 ng/mL rCTGF could increase the expression of p-ERK1/2. CONCLUSIONS: rCTGF may promote the proliferation and differentiation of human dental pulp cells through activating ERK1/2 signaling pathway.

Key words: Recombinant connective tissue growth factor, Human dental pulp cells, Cell proliferation, Cell differentiation, ERK1/2 signaling pathway

中图分类号:

R781.3

曹颖, 彭伟伟, 王璇, 伍甜甜, 杜嵘, 朱亚琴. 重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响[J]. 上海口腔医学, 2020, 29(1): 7-12.

CAO Ying, PENG Wei-wei, WANG Xuan, WU Tian-tian, DU Rong, ZHU Ya-qin. Effect of recombinant connective tissue growth factor on proliferation and odontogenic differentiation of dental pulp cells[J]. Shanghai Journal of Stomatology, 2020, 29(1): 7-12.

参考文献

[1] Gronthos S, Mankani M, Brahim J, et al.Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo[J]. Proc Nati Acad Sci USA, 2000, 97(25): 13625-13630.
[2] Wei X, Ling J, Wu L, et al.Expression of mineralization markers in dental pulp cells[J]. J Endod, 2007, 33(6): 703-708.
[3] Yongchaitrakul T, Pavasant P.Transforming growth factor-β1 up-regulates the expression of nerve growth factor through mitogen-activated protein kinase signaling pathways in dental pulp cells[J]. Eur J Oral Sci, 2010, 115(1): 57-63.
[4] Muromachi K, Kamio N, Matsumoto T, et al.Role of CTGF/CCN2 in reparative dentinogenesis in human dental pulp[J]. J Oral Sci, 2012, 54(1): 47-54.
[5] 江龙, 朱亚琴. 改良组织块法体外培养人牙周膜细胞和牙髓细胞[J]. 牙体牙髓牙周病学杂志, 2008, 18(4): 195-199.
[6] Kubota S, Takigawa M.Cellular and molecular actions of CCN2/CTGF and its role under physiological and pathological conditions[J]. Clin Sci, 2015, 128(3): 181-196.
[7] Xing X, Li Z, Yu Z, et al.Effects of connective tissue growth factor(CTGF/CCN2) on condylar chondrocyte proliferation, migration, maturation, differentiation and signalling pathway[J]. Biochem Biophys Res Commun, 2017, 495(1): 1447.
[8] Takigawa M.CCN2: a master regulator of the genesis of bone and cartilage[J]. J Cell Commun Signal, 2013, 7(3): 191-201.
[9] Aoyama E, Kubota S, Khattab HM, et al.CCN2 enhances RANKL-induced osteoclast differentiation via direct binding to RANK and OPG[J]. Bone, 2015, 73(1): 242-248.
[10] Perbal B.The concept of the CCN protein family revisited: a centralized coordination network[J]. J Cell Commun Signal, 2018, 12(1): 3-12.
[11] 隋国良, 彭依群, 何玉玲, 等. 重组人结缔组织生长因子对人成骨细胞增殖、分化及凋亡的影响[J]. 中华内分泌代谢杂志, 2008, 24(4): 360-363.
[12] Safadi FF, Xu J, Smock SL, et al.Expression of connective tissue growth factor in bone: Its role in osteoblast proliferation and differentiation in vitro and bone formation in vivo[J]. J Cell Physiol, 2003, 196(1): 51-62.
[13] Duan X, Ji M, Deng F, et al.Effects of connective tissue growth factor on human periodontal ligament fibroblasts[J]. Arch Oral Biol, 2017, 84: 37-44.
[14] Chen Y, Zhang Y, Ramachandran A, et al.DSPP Is essential for normal development of the dental-craniofacial complex[J]. J Dent Res, 2015, 95(3): 302-310.
[15] Nishida T, Nakanishi T, Asano M, et al.Effects of CTGF/Hcs24, a hypertrophic chondrocyte-specific gene product, on the proliferation and differentiation of osteoblastic cells in vitro[J]. J Cell Physiol, 2000, 184(2): 197-206.
[16] Chen C, Chen N, Lau LF.The angiogenic factors Cyr61 and connective tissue growth factor Induce adhesive signaling in primary human skin fibroblasts[J]. J Biol Chem, 2001, 276(13): 10443-10452.
[17] Kawaki H, Kubota S, Suzuki A, et al.Differential roles of CCN family proteins during osteoblast differentiation: Involvement of Smad and MAPK signaling pathways[J]. Bone, 2011, 49(5): 975-989.
[18] Kiwanuka E, Andersson L, Caterson EJ, et al.CCN2 promotes keratinocyte adhesion and migration via integrin α5β1[J]. Exp Cell Res, 2013, 319(19): 2938-2946.
[19] Luo Z, Kohli MR, Yu Q, et al.Biodentine induces human dental pulp stem cell differentiation through mitogen-activated protein kinase and calcium-/calmodulin-dependent protein kinase II pathways[J]. J Endod, 2014, 40(7): 937-942.
[20] Du R, Wu T, Liu W, et al.Role of the extracellular signal-regulated kinase 1/2 pathway in driving tricalcium silicate-induced proliferation and biomineralization of human dental pulp cells in vitro[J]. J Endod, 2013, 39(8): 1023-1029.
[21] 刘从娜. STAT3在CTGF调控人牙髓细胞分化中作用的研究[D]. 石家庄: 河北医科大学, 2015.

重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响

Effect of recombinant connective tissue growth factor on proliferation and odontogenic differentiation of dental pulp cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	陈祺, 谢非, 季颖颖, 魏魁杰, 任强. LLY-283靶向PRMT5抑制头颈部鳞癌增殖和转移的实验研究[J]. 上海口腔医学, 2022, 31(2): 142-147.
[2]	徐方芳, 蒋备战. DKK1对脂多糖感染人牙髓细胞生物学行为的影响[J]. 上海口腔医学, 2021, 30(4): 355-359.
[3]	王科, 彭国光, 谭玉莲, 何善志, 罗翠芬. miR-99a对口腔鳞癌增殖能力的影响[J]. 上海口腔医学, 2021, 30(1): 44-49.
[4]	郑晶, 李天客, 包阳, 张素欣. Spindly和Bub3在口腔鳞状细胞癌中的表达及意义[J]. 上海口腔医学, 2020, 29(5): 528-532.
[5]	冯小东, 易文, 殷卫红. 过表达miR-218对舌鳞癌细胞系SCC-4和SCC-9的增殖和侵袭的影响[J]. 上海口腔医学, 2019, 28(4): 373-377.
[6]	张漫, 李昊, 徐婷, 刘杰, 谈飞. 含锶交联藻酸钠凝胶对前成骨细胞黏附和增殖的影响[J]. 上海口腔医学, 2019, 28(2): 123-127.
[7]	彭伟伟, 戴兆威, 曹颖, 韩俊力, 朱亚琴. 血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响[J]. 上海口腔医学, 2019, 28(1): 1-5.
[8]	王宇琛, 王辰, 刘娜, 蔡川, 李伟, 徐璐璐. TNF-α对人脱落乳牙牙髓干细胞促破骨细胞形成能力的影响[J]. 上海口腔医学, 2018, 27(5): 449-454.
[9]	林田, 赵文青, 陆彦玲, 刘玉莹, 包丽荣, 吴煜. 人脐带间充质干细胞与牙髓细胞体外共培养对细胞生物学的影响[J]. 上海口腔医学, 2018, 27(4): 365-369.
[10]	李莉芬, 文扬, 江龙, 朱亚琴. 衣霉素诱导牙髓细胞内质网应激模型的建立[J]. 上海口腔医学, 2018, 27(2): 135-138.
[11]	高锦瑜, 余小琴, 王军强. 周期性牵张应力作用下KLF5对人牙周膜细胞增殖和成骨分化的影响[J]. 上海口腔医学, 2018, 27(1): 28-33.
[12]	包丽荣, 赵文青, 林田, 陆彦玲, 吴煜. BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化[J]. 上海口腔医学, 2017, 26(5): 476-483.
[13]	李凤霞, 王俊, 马玉云. CCN3调控牙周膜成纤维细胞的增殖和凋亡[J]. 上海口腔医学, 2017, 26(5): 504-509.
[14]	李晓明, 袁长永, 朱绍跃, 刘宗响, 王鹏来. 基因转染EphB4对脱落乳牙牙髓干细胞成骨分化的影响[J]. 上海口腔医学, 2017, 26(4): 399-403.
[15]	刘秉尧, 曹罡, 董震, 陈伟, 徐金科, 张森林, 翁志强. 沉默Grb7对口腔鳞癌细胞Cal27生物学行为的影响[J]. 上海口腔医学, 2016, 25(6): 688-693.