原花青素对人牙髓干细胞的增殖及成骨分化作用的研究  

Effect of proanthocyanins on the proliferation and osteogenic differentiation of human dental pulp stem cells

作　　者：刘亚军 丁如愿 杨刚 徐建光[1,2] 王元银[1,2] Liu Yajun;Ding Ruyuan;Yang Gang(Stomatological College of Anhui Medical University,Hefei 230032;Dept of Oral and Maxillofacial Surgery,The Affiliated Stomatological Hospital of Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学口腔医学院,合肥230032 [2]安徽医科大学附属口腔医院口腔颌面外科,合肥230032

出　　处：《安徽医科大学学报》2021年第11期1728-1733,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81271162);安徽省重点研究和开发计划项目(编号:201904a07020062)。

摘　　要：目的探讨原花青素(PAC)对人牙髓干细胞(hDPSCs)增殖及成骨分化能力的影响。方法改良组织块消化法分离培养hDPSCs,流式细胞术鉴定细胞表型,诱导染色鉴定分化潜能。不同浓度的PAC(0、4、8、16、32μmol/L)刺激hDPSCs后,通过CCK-8法检测hDPSCs的增殖,细胞骨架染色检测hDPSCs的细胞形态,碱性磷酸酶(ALP)活性检测hDPSCs的ALP活性,荧光定量PCR(q-PCR)检测成骨相关基因核心转录因子2(Runx-2)、骨形成蛋白2(Bmp-2)和Ⅰ型胶原(Col-1)的mRNA表达水平。结果体外成功培养hDPSCs并进行鉴定;CCK-8检测和细胞骨架染色结果显示,0~16μmol/L PAC对hDPSCs形态和增殖没有影响,32μmol/L PAC对hDPSCs增殖有抑制作用;ALP染色和定量检测均显示8μmol/L PAC组ALP活性较对照组(PAC 0μmol/L)高;q-PCR结果显示,与对照组相比,8μmol/L PAC诱导7 d后,成骨相关基因Runx-2、Bmp-2、Col-1的表达均升高。结论PAC对hDPSCs的成骨分化有促进作用。Objective To investigate the effect of proanthocyanins(PAC)on the proliferation and osteogenic differentiation of human dental pulp stem cells(hDPSCs).Methods hDPSCs were isolated and cultured in vitro,and the cells were treated with different concentrations of PAC(0,4,8,16,32μmol/L),then the proliferation of hDPSCs was detected by CCK-8,the cell morphology of hDPSCs was detected by cytoskeleton staining,the alkaline phosphatase(ALP)activity was detected by ALP staining,and the mRNA expression levels of osteogenic related genes like core transcription factor 2(Runx-2),bone morphogenetic protein 2(Bmp-2)andⅠcollagen type(Col-1)were detected by fluorescence quantitative PCR(q-PCR).Results hDPSCs were successfully cultured and identified.CCK-8 assay and cytoskeleton staining showed that 0-16μmol/L PAC had no effect on the morphology and proliferation of hDPSCs,but 32μmol/L PAC inhibited the cell proliferation significantly.ALP staining and quantitative detection showed 8μmol/L PAC was higher than control group in ALP activity.PCR showed that the expression of 8μmol/L PAC induction osteogenesis related genes in Runx2,Bmp2,Col-1 were significantly up-regulated.Conclusion PAC can promote the osteogenic differentiation of dental pulp mesenchymal stem cells.

关 键 词：原花青素 人牙髓干细胞 增殖 成骨分化 

分 类 号：R781.4[医药卫生—口腔医学]