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1、口腔生物醫(yī)學2014年6月(第5卷第2期)OralBiomedicine,Vo1.5,No.2,Jun.2014·75·[文章編號]1674—8603(2014)02—0075—03中間普氏菌培養(yǎng)上清對牙齦成纖維細胞RANKL/OPG表達的影響楊迷芳,楊天,王子露,王琰玲,吳友衣(南京醫(yī)科大學口腔醫(yī)學研究所,江蘇南京210029)[摘要]目的:探討中間普氏菌培養(yǎng)上清對牙齦成纖維細胞核因子一KB受體活化因子配體(receptoractivatorfornuc]earfactor—KBligand,RANKL)、骨保護素(osteoprotege
2、rin,OPG)表達的影響。方法:將系列濃度的中間普氏菌培養(yǎng)上清作用于牙齦成纖維細胞,顯微鏡下觀察細胞的變化。實時定量RTPCR法和Westernblot法分別檢測RANKL、OPGmRNA及蛋白水平的表達,并計算RANKL/OPG的比值。結果:牙齦成纖維細胞的數(shù)量隨中間普氏菌培養(yǎng)上清濃度的增加而減少,50g/mL時,顯微鏡下沒有活細胞存在。在mRNA水平上和蛋白水平上,RANKL/OPG的比值在12.5g/mL處理組均高于對照組,差異具有統(tǒng)計學意義(P<0.o1)。結論:中間普氏菌培養(yǎng)上清可誘導人牙齦成纖維細胞表達RANKL和OPG,破壞R
3、ANKL/OPG比例的平衡,影響破骨細胞分化的細胞因子微環(huán)境,在破骨細胞分化和牙周炎骨吸收中起著一定的調節(jié)作用。[關鍵詞]中間普氏菌;牙齦成纖維細胞;核因子一-(B受體活化因子配體;骨保護素[中圖分類號]R780.2[文獻標識碼]A[doi]10.3969/j.issn.1674—8603.2014.02.005EfectofPrevotellaintermediasupernatantontheexpressionofRANKLandOPGinhumangingivalfibroblastsYANGMifang,YANGTian,WANGZ
4、ilu,WANGYanling,UYounong.(InstituteofStomatology,NangMedicalUniversity,Nanjing210029,China)Correspondingauthor:YANGMng,Email:yangmng@qq.com,Tel:0086—25—86862025[Abstract]Objective:ToevaluatetheeffectsofPrevoteUaintermediasupernatantontheexpressionofreceptoractivatorofNF—KBl
5、igand(RANKL)andosteoprotegerin(OPG)inhumangingivalfibroblasts(HGFs).Methods:TheHGFsweretreatedwithgradientPrevotellaintermediasupernatant.HGFswereobservedunderthemicroscope.Thereal—timeRT-PCRandWesternblotanalysiswereusedfortestingtheexpressionofRANKLandOPG,andtheRANKL/OPGr
6、atiowascalculated.Results:Thenumberofcellsdecreasedwithincreaseofthesupernatantconcentration.At50Ixg/mL,nocellsurvivalcouldbeobservedunderthemicroscope.TheratioofRANKL/OPGwasincreasedinthecellstreatedwith12.5g/mLofPrevotellaintermediasupernatant(P<0.01).Conclusions:Prevo-te
7、llaintermediasupernatantcou]ddisruptthehomeostasisofRANKL/OPG,destroythemicroenvironmentofosteoclastdiferentiation.Prevotellaintermediasupernatantmayplayaregulatoryroleinosteoclastdifferentiationandboneresorptioninperiodontitis.[Keywords]Prevotellaintermedia;Gingivalfibrobl
8、asts;ReceptoractivatorofNF—KBligand;Osteoprotegerin牙齦成纖維細胞(humangingivalfibroblast