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ID:18172245
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页数:5页
时间:2018-09-15
《微管蛋白破坏对软骨细胞代谢功能的影响》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、万方数据·1036·主垡匡堂盘鲞垫!!生堡旦!!旦笠!!鲞筮!!翅堕坐丛鲤!£h塾堡:垒旦堕!!:垫!!:!丛!!:堡垒!i微管蛋白破坏对软骨细胞代谢功能的影响郭恒段王平李琦曹晓明王磊卫小春【摘要】目的探讨微管蛋白破坏对体外关节软骨细胞代谢功能的影响。方法2月龄新西兰白兔8只,处死后取双膝关节全层软骨,采用常规0.4%链酶菌蛋白酶和0.025%II型胶原酶依次消化为软骨细胞培养3d贴壁后,分为对照组和实验组,对照组继续用原代培养基(90%DMEM/F12+10%胎牛血清)培养,实验组在原代培养基中加入微管蛋白破坏剂秋水仙素(终浓度
2、为0.1p.mol/L)。加药后第l、2天用Annexin—V/PI流式细胞术检测软骨细胞早期凋亡率,加药后第6天细胞爬片HE染色观察细胞形态的改变,加药后第3、6.9天取细胞用实时定量荧光反转录聚合酶链式反应法测定软骨细胞Ⅱ型胶原、蛋白多糖以及MMP-13mRNA的表达量,同时在加药后第3、6、9天取细胞上清液,用ELISA法和阿尔新蓝法检测各组上清液中Ⅱ型胶原和蛋白多糖的含量。结果实验组第2天软骨细胞早期凋亡率明显高于对照组(P<0.05);与对照组比较,实验组第6天软骨细胞呈不规则多角形,胞核深染且分裂象增多,细胞基质减少,
3、实验组第3、6、9天软骨细胞Ⅱ型胶原和蛋白多糖mRNA表达量较对照组均明显降低(P<0.05),第6、9天实验组软骨细胞MMP·13mRNA表达较对照组明显增高(P4、thehomeostasisofarticularcartilagechondrocytesGUO胁昭,DUANWang-ping,ⅡQi,CAOXiao—ming,WANGLei,WEIXiao—chun.DepartmentofOrthopedics,SecondHospitalofShanxiMedicalUnivers毋;Shanxi研LaboratoryofBone&sogTissueInjuryRepair,Taiyuan030001,ChinaCorrespondingauthor:WEIXiao—chun,Emai5、l:weixiaochun06@yahoaCOllhcn【Abstract】0bjecfiveToinvestigatetheinfluenceoftubulindisassemblyontheinvitrometabolismofarticularehondrecytes.MethodsEightNewZealandrabbitsaged2momhaweresacrificedbyairembolism.nefull-thicknesscartilageswereharvestedfrombothkneesundersteril6、econditions.Then0.4%pronaseand0.025%1IcoUagenagewereusedtodigestforprimarychondrocytes.r11地ceHswereculturedfor3daysafterattachmentandtIlendividedintothecontrolandexperimentalgroups.Thecontrolgroupcontinuedculturingwithprimarymediumandtheexperimentalgroupwithcolchicine7、,atubulindestructiveagent.atafinalconcentrationof0.1p,mol/LAtDaysl&2,theearlyapeptosisoftwocellgroupsWaSassayedwithphosphatidylserine(AnnexinV).AtDay6,themorphologicalchangesofcellswereobservedbyhematoxylinandeosinstaining.AtDays3,6&9,theexpressionlevelsoftypeⅡcollage8、n,proteogbrcarlandMMP(matrixmetalloproteinase).13mBNAwereIneRsuredbyrealtimequantitativefluorescentmver孵Ⅱanscription.polymer
4、thehomeostasisofarticularcartilagechondrocytesGUO胁昭,DUANWang-ping,ⅡQi,CAOXiao—ming,WANGLei,WEIXiao—chun.DepartmentofOrthopedics,SecondHospitalofShanxiMedicalUnivers毋;Shanxi研LaboratoryofBone&sogTissueInjuryRepair,Taiyuan030001,ChinaCorrespondingauthor:WEIXiao—chun,Emai
5、l:weixiaochun06@yahoaCOllhcn【Abstract】0bjecfiveToinvestigatetheinfluenceoftubulindisassemblyontheinvitrometabolismofarticularehondrecytes.MethodsEightNewZealandrabbitsaged2momhaweresacrificedbyairembolism.nefull-thicknesscartilageswereharvestedfrombothkneesundersteril
6、econditions.Then0.4%pronaseand0.025%1IcoUagenagewereusedtodigestforprimarychondrocytes.r11地ceHswereculturedfor3daysafterattachmentandtIlendividedintothecontrolandexperimentalgroups.Thecontrolgroupcontinuedculturingwithprimarymediumandtheexperimentalgroupwithcolchicine
7、,atubulindestructiveagent.atafinalconcentrationof0.1p,mol/LAtDaysl&2,theearlyapeptosisoftwocellgroupsWaSassayedwithphosphatidylserine(AnnexinV).AtDay6,themorphologicalchangesofcellswereobservedbyhematoxylinandeosinstaining.AtDays3,6&9,theexpressionlevelsoftypeⅡcollage
8、n,proteogbrcarlandMMP(matrixmetalloproteinase).13mBNAwereIneRsuredbyrealtimequantitativefluorescentmver孵Ⅱanscription.polymer
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