【作者】 汪昌宁； 杨芳； 胡英英； 哈珊珊；

【Author】 WANG Chang-ning;YANG Fang;HU Ying-ying;HA Shan-shan;The State Key Laboratory Breeding Base of Basic Science of Stomatology & Key Laboratory of Oral Biomedicine Ministry of Education,School of Stomatology,Wuhan University;Department of Periodontology,Hospital of Somatology,Wuhan University;

【机构】 武汉大学口腔医学院·湖北省口腔基础医学重点实验室-省部共建国家重点实验室培育基地·武汉大学口腔生物医学教育部重点实验室； 武汉大学口腔医院牙周科；

【摘要】 牙周炎以牙龈软组织炎症和牙槽骨等骨组织破坏吸收为显著表现,是引起成人失牙的首要原因。PPARγ可调节糖脂代谢,细胞增殖、分化、凋亡,参与炎症、肿瘤、动脉粥样硬化、糖尿病、肥胖症等疾病过程。PPARγ可抑制炎症进程,但是对成骨的影响存在争议。在非炎症状态下,PPARγ促进间充质干细胞成脂分化,抑制其成骨分化;但在种植体周围炎的成骨细胞中其可促进成骨,因此PPARγ对成骨影响可能和细胞类型和所处环境有关。PPARγ在炎症和非炎症状态下对人牙周膜细胞成骨影响及其机制目前仍不清楚。目的:探究PPARγ在炎症和非炎症状态下对人牙周膜细胞成骨影响和机制。方法:组织块法培养人牙周膜细胞,在炎症和非炎症状态下,采用PPARγ激动剂(罗格列酮)和抑制剂(GW9662)刺激细胞,矿化诱导3,7,14天。碱性磷酸酶(ALP)活性和染色试剂盒检测ALP含量变化;实时荧光定量PCR检测RUNX2、OCN、OPG、RANKL、ColI、BMP2、ALP基因的表达;茜素红染色观察矿化结节的形成;免疫印迹法检测β-catenin在总蛋白和胞核蛋白中的变化。结果:在炎症和非炎症状态下,PPARγ激动剂可提高人牙周膜细胞ALP活性和促进矿化结节的生成,降低总蛋白和胞核蛋白中β-catenin的含量(P<0.05),而PPARγ抑制剂无法逆转β-catenin的含量变化;较非炎症状态,炎症状态时,PPARγ激动剂促进ALP、OPG、BMP2基因表达(P<0.05)。结论:在炎症和非炎症状态下,PPARγ激动剂促进人牙周膜细胞的成骨,但不依赖于β-catenin信号通路,为临床牙周炎治疗提供新的治疗思路。更多还原

【Abstract】 Periodontitis is the main reason contributes to teeth loss of adults and is characterised by soft tissue inflammation and bone tissue(alveolar bone) absorption.PPARγ regulates glucolipid metabolism,cell proliferation,differentiation and apoptosis;and participates in various diseases process including inflammation,cancer,atherosclerosis,diabetes and obesity.It restrains inflammatory process,but the effect in osteogenesis is controversial.In noninflammatory situation,PPARγ facilitates adipogenic differentiation and inhibits osteogenesis in mesenchymal cells differentiation,but it expedites osteogenesis in osteoblast of periimplantitis,so the influence of PPARγ on osteogenesis may related to cell types and its living environment.The influence and mechanism of PPARγ on osteogenesis in hPDLCs in inflammatory and noninflammatory situation still remains indistinct.Objective:Explore the influence and mechanism of PPARγ on osteogenesis in hPDLCs in inflammatory and noninflammatory situation.Methods:Culturing hPDLCs,stimulating cells in inflammatory and noninflammatory environment with PPARγ agonist(rosiglitazone) and anagonist(GW9662) for a period of 3,7 and 14 days seperately.Detecting ALP activity with ALP activity detection kit and finding out the gene expression of RUNX2,OCN,OPG,RANKL,Col I,BMP2 and ALP using Quantitative realtime PCR;Observing the mineralization nodules with alizarin red staining method and examining the percentage changes of β-catenin in total protein and nucleic protein through western blot analysis.Results:PPARγ agonist improves ALP activity,promotes mineralization nodules and reduces β-catenin content in total protein and nucleic protein either in inflammatory status or noninflammatory status(P<0.05),but PPARγ agonist can’t reverse the change of β-catenin.Compared to noninflammatory situation,PPARγ agonist can accelerate gene expression of ALP,OPG and BMP2 in inflammatory situation(P<0.05).Conclusions:PPARγ agonist contributes to the osteogenesis of hPDLCs in both inflammatory and noninflammatory situation without relying on β-cateninsignaling pathway,and this provides new ideas for clinical treatment of periodontitis.更多还原