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1、sirtuin1促进牙周膜干细胞骨向分化的初步研宄中德山1刘亚蕊2张清彬3(通讯作者)唐伟峰4(1湖北省荣军医院口腔科430079)(2广州医科大学附属口腔医院儿童口腔科510140)(3广州医科大学口腔医学重点实验室510140)(4北京大学口腔医院第三门诊部100120)【摘要】目的研究蛋白酶sirtuin1在牙周膜干细胞骨向分化中的作用。方法筛选人因正畸而拔除的前磨牙,获取牙周膜组织做细胞培养;将细胞按照实验组、对照组进行分组。实验组h加入Sirtuinl激活剂白藜芦醇使其工作浓度为1mM,5mM,10mM
2、,实验组II:加入Sirtuinl抑制剂烟酰胺使其终末工作浓度为1mM,5mM,10mM,对照组为空白对照。获取培养后细胞做蛋白酶Sirtuin2和各种细胞骨向分化标志物检测:West-blotting检测Sirtuinl和各种骨向分化标志物蛋白的表达;RT-PCR检测骨向分化标志物ALP,骨桥蛋白,骨钙蛋白和骨涎蛋白。结果通过检测细胞的蛋白酶Sirtuinl和骨向分化标志物ALP,骨桥蛋白,骨钙蛋白和骨涎蛋白的mRNA,表明随着加入白芦藜醇的剂量的差异而岀现梯度的上调;而随着烟酰胺的浓度的升高而出现梯度的下调。
3、结论sirtuinl可以有效的促进牙周膜干细胞骨向分化,从而促进牙槽骨再牛.修复重建,其有良好的临床应用前景。【关键词】sirtuin1细胞培养牙周膜干细胞成骨作用【中图分类号】R78【文献标识码】A【文章编号】2095-1752(2013)19-0048-03Theroleofsirtuin1inpromotionofosteoblasticdifferentiationonperiodontalligamentstemcellsXuJinhanlLiuYarui2,ZhangQingbin3,TangWeif
4、eng4(1.Generaldepartment,schoolandhospitalofstomatology,Guangzhoumedicaluniversity,GuangdongProvince,China,5101402.Departmentofpediatricdentistry,Guangzhoumedicaluniversity,GuangdongProvince,China,510140;2.Keylabofstomatolgy,schoolandhospitalofstomatology,Gua
5、ngzhoumedicaluniversity,GuangdongProvince,China,510140;3.Thethirddentalclinic,hospitalofstomatology,Pekinguniversity,Beijingcity,China,100120)【Abstract】Objective:ToidentifytheroleofSIRT1inosteogenicdifferentiationofperiodontalligamentstemcells.Method:weinvest
6、igatedtheeffectsofoverexpressingandunderexpressingSIRT1onthedifferentiationofperiodontalligamentstemcells,ExpressionofSIRT1andosteoblasticdifferentiationmarkerswasassessedbyRT-PCR,andwesternblotting.Result:ActivationofSIRT1usingresveratrolstimulatedosteoblast
7、icdifferentiationinadose-dependentmanner,asassessedbytheexpressionofmRNAsencodingalkalinephosphatase,osteopontin,osteocalcin,osterixandRunx2,andinducedcalciumdeposition.Incontrast,inhibitionofSIRT1usingnicotinamidesuppressedmineralizationandtheexpressionofost
8、eoblastmarkermRNAs..Conclusion:ItisverifiedthatSIRT1isapotentregulatorofdifferentiationofhumanperiodontalligamentcellsandmayhaveclinicalimplicationsforperiodontalboneregeneration.【Keyword