细胞骨架及相关蛋白在人牙周韧带细胞成骨分化过程中的表达模式  

Expression of the cytoskeleton proteins and cytoskeleton-associated proteins in human periodontal ligament cell osteogenic differentiation

作　　者：吴莉萍[1] 韦曦[1] 凌均棨[1] 刘路[1] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.口腔医学研究所,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2008年第5期8-12,共5页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(30872879)

摘　　要：目的探讨人牙周韧带细胞(PDLCs)成骨分化过程中细胞骨架及相关蛋白的表达模式及其可能的功能作用。方法酶消化法培养PDLCs,免疫细胞化学染色检测vimentin的表达;取诱导前(对照组)及矿化诱导7、14和21d的PDLCs,实时荧光定量RT-PCR检测vimentin、actin、caldesmon(CaD)、tropomyosin(Tm)和annexinA4mRNA的表达变化并进行统计分析,Westernblot检测蛋白表达。结果PDLCs表达丰富的vimentin;Vimentin、actin、CaD和TmmRNA的表达在诱导7d组下调,诱导14d组和21d组逐渐上调;AnnexinA4mRNA的表达在诱导7d组表现为上调,诱导14d组和21d组逐渐下调;Vimentin的mRNA表达在各组间差异均有统计学意义(P<0.05);CaD和Tm在诱导7d组与14d组之间的mRNA表达差异无统计学意义(P>0.05),其余各组间差异均有统计学意义(P<0.05);Actin和annexinA4在对照组与诱导21d组之间的mRNA表达差异无统计学意义(P>0.05),其余各组间差异均有统计学意义(P<0.05);Western blot检测蛋白表达显示类似的表达趋势。结论在PDLCs成骨分化过程中,一组细胞骨架及细胞骨架蛋白相关蛋白呈现相似的表达变化,提示其参与调节PDLCs成骨分化。Objective The expression of the cytoskeleton and cytoskeleton-associated proteins was investigated to explore its possible function in human periodontal ligament cells （PDLCs） osteogenic differentiation. Methods PDLCs were digested and cultivated by a solution containing eollagenase type I and dispase. The expression of vimentin in PDLCs were investigated by immunoeytochemistry. PDLCs were proceeded to osteogenic induction for 7-, 14- and 21-days respectively, and cells before induction served as controls （0 d）. The expressions of vimentin, aetin, caldesmon（CaD）, tropomyosin（Tm） and annexin A4 mRNAs were investigated by real-time fluorescence quantitative RT-PCR analysis, and statistical analysis was performed to compare the differences of mRNA expression levels among cell samples collected at different time points. The expression of proteins was investigated by western blot. Results Positive staining for vimentin was observed in PDLCs. The mRNA expressions of vimentin, actin, CaD and Tm decreased in the 7-day induction group but increased in the 14- and 21-day induction groups. The mRNA expressions of annexin A4 increased in the 7-day induction group but decreased in the 14- and 21-day induction groups. There are significant differences between any two groups for vimentin mRNA expressions （P 〈0.05）. There are significant differences between other two groups for CaD and Tm mRNA expressions （P 〈 0.05） except between the 7-day and 14-day induction groups （P 〉 0.05）. There are significant differences between other two groups for aetin and annexin A4 mRNA expressions （P 〈 0.05） except between control group and 21 day induction groups （P 〉 0.05）.The proteins show analogously temporal expression of mRNAs. Conclusions PDLCs show analogously temporal expression of the cytoskeleton and cytoskeleton-associated proteins as differentiate into cementoblast/osteoblast-like cells in vitro, it is indicated those proteins may play a regulatory role in PDLCs osteogenic differentiation

关 键 词：牙周韧带细胞 成骨分化 细胞骨架蛋白 

分 类 号：R78[医药卫生—口腔医学]