中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (28): 4475-4480.doi: 10.3969/j.issn.2095-4344.0390

• 口腔组织构建 oral tissue construction • 上一篇    下一篇

中药黄芩苷干预人牙周膜成纤维细胞的作用及分子机制

李丽华1,贾艾敏2,杨明辉2,刘  震1,王丽恒2,蒋  红2,邱  亚2   

  1. 川北医学院附属医院,1口腔科, 2医学研究中心,四川省南充市  637000
  • 收稿日期:2018-04-06 出版日期:2018-10-08 发布日期:2018-10-08
  • 通讯作者: 邱亚,硕士,副主任医师,川北医学院附属医院医学研究中心,四川省南充市 637000
  • 作者简介:李丽华,女,1977年生,四川省广安市人,汉族,2007年重庆医科大学毕业,硕士,副教授,主要从事口腔基础和临床的相关研究。
  • 基金资助:

    四川省卫计委科研课题(16PJ132);四川省教育厅课题(14ZB0197)

Effect of baicalin on human periodontal ligament fibroblasts and the underlying molecular mechanism

Li Li-hua1, Jia Ai-min2, Yang Ming-hui2, Liu Zhen1, Wang Li-heng2, Jiang Hong2, Qiu Ya2   

  1. 1Department of Stomatology, 2Medical Research Center, Affiliated Hospital of Northern Sichuan Medical College, Nanchong 637000, Sichuan Province, China
  • Received:2018-04-06 Online:2018-10-08 Published:2018-10-08
  • Contact: Qiu Ya, Master, Associate chief physician, Medical Research Center, Affiliated Hospital of Northern Sichuan Medical College, Nanchong 637000, Sichuan Province, China
  • About author:Li Li-hua, Master, Associate professor, Department of Stomatology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
  • Supported by:

    the Scientific Research Project of Health and Family Planning Commission of Sichuan Province, No. 16PJ132; the Project of Education Department of Sichuan Province, No. 14ZB0197

摘要:

文章快速阅读:

文题释义:
黄芩苷:是从植物黄芩的干燥根中提取分离出来的一种黄酮类化合物,具有多种药理作用,如抗菌、消炎、减少抗原抗体反应和化学介质的释放量、抑制变态反应和清除超氧自由基等。研究表明,黄芩苷能够在一定程度上抑制基质金属蛋白酶的活性,能够促进成纤维细胞的增殖和胶原蛋白的合成。
骨形态发生蛋白2:近年来基因重组的骨形态发生蛋白2应用的安全性和有效性得到了充分的证实,美国FDA也于2002年7月批准重组骨形态发生蛋白2应用于脊柱融合。基于骨形态发生蛋白的安全性和高效诱导成骨活性被越来越多的实验所证实并进入临床试验阶段。
摘要
背景:
黄芩苷作为一种重要的中药提取物,近年来的应用被越来越多的研究所提及。实验表明黄芩苷可以促进人牙周膜成纤维细胞的增殖并向成骨分化,但具体分子机制尚不清楚。
目的:研究中药提取物黄芩苷对人牙周膜成纤维细胞的增殖促进作用及潜在的分子机制。
方法:通过细胞原代培养及光学显微镜下观察人牙周膜成纤维细胞的形态。CCK8法检测黄芩苷对人牙周膜成纤维细胞增殖的促进作用;采用实时定量PCR分析黄芩苷对增殖细胞核抗原表达的影响;实时定量PCR以及流式细胞术分析黄芩苷对细胞凋亡的抑制作用;Western blot及实时定量PCR分析黄芩苷对细胞中骨形态发生蛋白2蛋白与mRNA表达的影响;加入骨形态发生蛋白2抗体来检测黄芩苷对细胞增殖凋亡的作用以及骨形态发生蛋白2表达的变化。
结果与结论:①与对照组相比,黄芩苷能够明显促进人牙周膜成纤维细胞增殖,并上调增殖细胞核抗原的mRNA表达水平;②与对照组相比,黄芩苷能够上调Bcl-2 mRNA表达水平,下调Bad mRNA表达水平,流式细胞术结果也显示黄芩苷能够抑制细胞的凋亡;③黄芩苷能够浓度与时间依赖性地上调骨形态发生蛋白2蛋白与mRNA水平;加入骨形态发生蛋白2中和抗体可减弱黄芩苷的调控作用;④结果表明,黄芩苷能够促进人牙周膜成纤维细胞的增殖,其发挥作用的机制可能与上调骨形态发生蛋白2表达有关。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0001-7215-1324(李丽华)

关键词: 黄芩苷, 人牙周膜成纤维细胞, 增殖, 凋亡, 骨形态发生蛋白2, 牙周病, 成骨分化

Abstract:

BACKGROUND: Baicalin, as an important traditional Chinese medicine extract, has been extensively studied. Baicalin has been shown to promote the proliferation of human periodontal ligament fibroblasts (HPLFs) and differentiation into osteocytes, but the underlying molecular mechanism remains unclear.
OBJECTIVE: To study the effect of baicalin on the proliferation of HPLFs and the underlying molecular mechanism.
METHODS: Morphology of HPLFs was observed by primary cell culture and light microscopy. The growth-promoting effect of baicalin on HPLFs was detected by cell counting kit-8 assay. The effect of baicalin on the expression of proliferating cell nuclear antigen was detected by qRT-PCR. The inhibitory effect of baicalin on cell apoptosis was detected by qRT-PCR and flow cytometry. The level of bone morphogenetic protein 2 (BMP2) under baicalin treatment was tested by western blot assay and qRT-PCR. BMP2 antibody was employed to detect the influence of baicalin on the proliferation and apoptosis of HPLFs and the expression of BMP2 in HPLFs.
RESULTS AND CONCLUSION: Compared with the control group, baicalin could significantly promote the proliferation of HPLFs, and up-regulate the mRNA expression level of proliferating cell nuclear antigen. Compared with the control group, baicalin could significantly up-regulate the mRNA level of Bcl-2 and down-regulate the mRNA level of Bad. Flow cytometry also showed that baicalin could inhibit cell apoptosis. Baicalin could up-regulate the protein and mRNA levels of BMP2 in a time- and concentration-dependent manner. The regulatory effect of baicalin was decreased after addition of BMP2 antibody. In summary, baicalin promotes proliferation of HPLFs probably via up-regulating BMP2 level.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Drugs, Chinese Herbal, Periodontal Ligament, Cell Proliferation, Bone Morphogenetic Proteins, Tissue Engineering

中图分类号: