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1、唑来膦酸对破骨细胞生成及Syk基因表达的影响王宏利，李鹏，刘强，董伟，戚孟春，李金源（063000唐山，河北联合大学口腔医学院口腔颌面外科教研室请补充具体科室、教研室等）[摘要]目的探讨唑来膦酸(zoledronateacid，ZOL)在共培养体系中对脾酪氨酸激酶(spleentyrosinekinase，Syk)基因表达及破骨细胞(osteoclast，OC)生成的影响。方法体外将小鼠颅骨成骨细胞与单核巨噬细胞RAW264.7共培养，将细胞分为2组：对照组及5×10-7mol/LZOL处理24h组（ZOL组）。应
2、用TRAP染色、牙本质吸收陷窝检测OC生成及骨吸收情况；Real-timePCR、Westernblot、免疫荧光化学检测Syk基因表达写明时间点。结果两组细胞均有TRAP染色阳性多核OC生成，并在牙本质磨片上形成吸收陷窝；但OC生成及吸收陷窝的数目和体积ZOL组均显著少于对照组(P<0.01)。SykmRNA及蛋白水平在ZOL组也显著下降(P<0.01)。免疫荧光检测显示，Syk在对照组细胞质内强表达，并在细胞周缘形成肌动蛋白环样结构；而ZOL组Syk表达明显减弱，肌动蛋白环样结构消失。结论ZOL可显著抑制共培养
3、体系中OC生成和骨吸收功能，这可能与其对Syk基因表达的抑制有关。[关键词]唑来膦酸；破骨细胞生成；脾酪氨酸激酶；抗酒石酸酸性磷酸酶[中图法分类号][文献标志码]ATheeffectofzoledronateacidonosteoclastogenesisandgeneexpressionofSpleentyrosinekinaseinaosteoclast-osteoblastco-culturesysteminvitroWamgHongli,Lipeng,LiuQiang,DongWei,QiMengchun,
4、LiJinyuan(CollegeofStomatology,HebeiUnitedUniversity,Tangshan,HebeiProvince,063000,China)[Abstract]ObjectiveToexploretheeffectofzoledronateacid(ZOL)ongeneexpressionofspleentyrosinekinase(Syk)andosteoclastogenesisinanosteoclast-osteoblastco-culturesystem.Method
5、sMicecalvarialosteoblastsandRAW264.7cellswereco-culturedandthecellsweredividedinto2groups.GroupAservedascontrolandgroupBreceivedZOLtreatment(5×10-7mol/L)for24h.TRAPstaininganddentinresorptionlacunaeexaminationwereusedtodetectosteoclastformationandresorptionfun
6、ction,whileReal-timePCR,WesternblotandimmunofluorescentchemistrywereappliedtoexamineSykgeneexpression.ResultsTRAPpositivecellsanddentinresorptionlacunaewereobservedinbothgroups.However,groupBshowedlessTRAPpositivemulti-nucleatedOCandresorptionlacunaewhencompar
7、edwithgroupA(p<0.01).SykmRNAandproteinwerealsosignificantlydown-regulatedingroupB(p<0.01).ImmunofluorescentexaminationshowedthatSykwasstronglyexpressedincytoplasmofgroupAandformedactin-likeringsatcellperiphery;however,itwasonlyweaklyexpressedingroupBandnoactin
8、-likeringsformed.ConclusionIntheco-culturesystem,ZOLcouldsignificantlyinhibitformationandresorptionfunctionofmulti-nucleatedosteoclastsandthismaybeattributed,atleastpartly,toitsinh
