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1、·102·現(xiàn)代口腔醫(yī)學雜志2013年第27卷第2期JModernStomatol,March2013,Vol27,No.2脂多糖刺激對牙周膜細胞培養(yǎng)上清中RANKL和OPG水平改變的影響武海春許堯生李彬【摘要】目的檢測脂多糖(LPs)刺激后,體外培養(yǎng)人牙周膜細胞分泌核因子一kB受體活化子配體(RANKL)和骨保護素(OPG)濃度的變化,探討其對牙槽骨吸收的直接影響。方法組織塊法原代培養(yǎng)正常人牙周膜細胞并進行來源鑒定。以lOmg/mlLPS刺激第4代細胞,在不同時間段(Oh,4h,8h,12h,24h)用酶聯(lián)免疫吸附實驗(ELISA)檢測RANKL和OPG在細胞培養(yǎng)上清中的含量
2、,并進行統(tǒng)計學分析。結果ELISA結果顯示正常牙周膜細胞培養(yǎng)上清中可檢測到少量RANKL和OPG,LPs刺激后RANKL的表達則隨時間延長而逐漸增強,且4h、8h、12h、24h時間點各實驗組均與對照組有顯著差異(P<0.01)。各實驗組之間,8h、12h兩時間點有顯著差異(P<0.05),其他時間點之間無顯著差異(P>0.05);r~LPS刺激后不同時間OPG的含量未見顯著變化(P>0.05)。未經(jīng)IJPs刺激的牙周膜細胞培養(yǎng)上清中可檢測到少量RANKL和OPG,但隨時間延長無顯著變化。結論體外培養(yǎng)牙周膜細胞在LPs刺激后一定時間內(nèi)RANKL~達增強,而OPG表達無明顯變化
3、?!娟P鍵詞】人牙周膜細胞脂多糖骨保護素核因子一kB受體活化因子配體ELISATheproductionofreceptoractivatorofNF—kBligandandosteoprotegerininculturesupernatantofhumanperiodontalligamentceHsstimulatedwithE.coliLPSWUHmchun,XUYaoshen,LIBin.TheSecondPeopleHospitalofHengshui,Hengshui053000【Abstract】objectiveToexaminetheproductionofr
4、eceptoractivatorofNF-kBligand(RANKL)andosteoprotegerin(OPG)intheculturesupernatantofhumanperiodontalligamentcellsafterstimulationwithE.colilip0p0lysaccharide(LPS),andtoelucidatetheroleofperiodontalligamentcellsinalveolarboneresorption.MethodsPrimaryhumanperiodontalligamentcellswerecultureda
5、ndidentifiedimmunologically.11}lefourthpassageofthecellswasstimulatedwithEcoli.LPSfor0h,4h,8h,12h,and24h,enzymelinkedimmunosorbentassay(ELISA)wasusedtoexaminetheconcentrationofsecretedRANKLandOPGinthesupernatant.ResultsAfterstimulationwithLPS,OPGconcentrationinthesupernatantexhibitednosigni
6、ficantchange,butRANKLconcentrationinthestimulatedgroupsincreasedgraduallyfrom4h,andpeakedat24h,statisticanalysisrevealedsignificantdifferenceforallthestimulatedgroupswhencomparedwiththecontrolgroups(P<0.O1);withinstimulatedgroups,8hand12halsorevealedasignificantdiference(P<0.05);Groupswitho
7、utLPSstimulationexhibitedalowlevelofRANKLandOPG,andhadnosignificantchangesatanyofthetimepointsexamined.ConclusionPeriodontalligamentcellsstimulatedwithLPSexpressedanincreasedlevelofRANKL,butOPGexpressionhavenosignificantchange.【Keywords】Humanperiodontall