胆碱受体介导调控乳牙牙髓干细胞破骨能力的研究 .pdfVIP

内源性配体SLURP1经α7亚型烟碱型乙酰胆碱受体介导调

控乳牙牙髓干细胞破骨能力的研究

蒋旸;汪璐璐;周志斐;张百泽;王军辉;王小竞

【摘要】AIM:Toinvestigatetheroleofα7nAChRandSLURP1in

differentiationandmaturationofoste-oclastsintheprocessofdeciduous

teethphysiologicalrootresorption.METHODS:SHEDsfromdifferentstages

ofde-ciduousteethphysiologicalrootresorptionandDPSCsfrom

permanantteethwereisolated,cultivatedandpurifiedbyen-zyme

digestionmethodandlimitingdilutionassay.Thecellswereidentifiedby

cellularmorphology,growthpattern,os-teogenicinduction,adipogenic

differentiationandflowcytometryanalysis.Geneandproteinexpressionof

α7nAChRwasdetectedbyquantitativereal-time

PCR,immunofluorescenceandsemiquantitativeopticaldensity

analysis.SLURP1andRANKL/OPGgeneexpressionwasdetectedby

quantitativereal-timePCR.RESULTS:Geneandproteinexpres-sionofα7

nAChRinSHEDsfromthemiddleandthefinalstageofphysiologicalroot

resorptionwashigherthanthosefromthestablestageandhigherthan

DPSCs(P0.05).SLURP1expressionwasthehighestinthemiddlestage,fol-

lowedbythefinalstage,permanentteethandthestablestageof

deciduousteeth.(P0.05).RANKL/OPGratioshowedthesimilarexpression

tendencytoSLURP1expression.CONCLUSION:SLURP1mayregulatethe

osteoclas-togenicdifferentiationofstemcellsfromhumanexfoliated

deciduousteethbybindingtoα7nAChR.%目的：初步探讨α7亚型烟碱型乙

酰胆碱受体（α7nAChR）及其内源性配体SLURP1在乳牙生理性根吸收过程中

调控破骨细胞分化成熟的可能作用。方法：分离培养并优选乳牙生理性根吸收不同

时期乳牙牙髓干细胞（SHEDs）及恒牙牙髓干细胞（DPSCs），经检测鉴定后，

分别采用实时定量PCR、免疫荧光染色及半定量光密度分析技术检测各组细胞中

α7nAChR基因及蛋白的表达差异；实时定量PCR检测各组细胞中SLURP1、经

典成骨／破骨相关因子RANKL、OPG基因的表达差异，并进行统计学分析。结

果：乳牙牙根吸收中期及晚期的SHEDs中α7nAChR基因和蛋白的表达水平显

著高于乳牙牙根稳定期SHEDs与恒牙DPSCs（P＜0．05）；SLURP1基因表达

水平以乳牙牙根吸收中期最高，晚期次之，恒牙再次之，乳牙牙根稳定期最低，各

组间P＜0．05。RANKL／OPG比值与SLURP1基因表达趋势一致（P＜

0．05）。结论：SLURP1与乳牙牙髓干细胞膜上的α7nAChR结合后，可通过

调节RANKL／OPG的表达比例而