中文摘要：

目的研究小RNA干扰靶向抑制Par-4基因表达对人骨髓间充质干细胞（hBMSCs）中CD2相关蛋白（CD2AP）表达的影响及意义。方法利用构建靶向抑制Par-4基因的SiRNA真核表达载体转染hBMSCs。采用实时荧光定量PCR检测Par-4基因的mRNA水平。采用谷氨酸诱导hBMSCg凋亡。采用流式细胞术检测hBMsCs凋亡百分率。Western blot检测hBMSCs CD2AP蛋白表达量。应用SPSS10．0软件进行统计学分析。结果1．Par-4-SiRNA可下调hBMSCsPar-4-mRNA水平，抑制率为88％（P〈0．01）。而非抑制的对照序列未见抑制作用（P〉0．05）。2．流式细胞术检测结果显示，与正常对照组比较，谷氨酸加常规hBMSCs组凋亡百分率为（59．12±5．43）％（P〈0．01）。而在Par-4-SiRNA转染的Par-4-SiRNA-hBMSCs组中，谷氨酸对hBMSCs的凋亡诱导作用被显著抑制，凋亡百分率显著下调为（39．49±5．01）％（P〈0．01）。3．Par-4-SiRNA转染的Par-4-SiRNA-hBMSCs组，谷氨酸对hBMSCs中CD2AP的下调作用被显著抑制，CD2AP蛋白的表达量显著上升（P〈0．01）。结论靶向抑制Par-4基因的表达能够拮抗谷氨酸诱导的hBMSCs凋亡及CD2AP表达的下调。

英文摘要：

Objective To explore the effects and significance of small interfering RNA against Par - 4 gene on expression of CD2 - associated protein （CD2AP） in human bone marrow mesenchymal stem cells （hBMSCs） exposed to glutamate. Methods Primary culture of hBMSCs were carried out and SiRNAs targeted Par - 4 gene （ Par - 4 - SiRNA） were chemically synthesized. Normal control, glutamate - treated group, glutamate and Par- 4 -siRNA treated group, glutamate and scrambled siRNA -treated group were divided. The expression of Par - 4 mRNA was determined by real - time quantitative polymerase chain reaction. Eukaryocytic expression vector was built and was transfected into hBMSCs and the cells were treated with glutamate. The apoptosis of hBMSCs was quantified by flow cytometry. Western blot was used to detect the protein levels of CD2AP. The data were analyzed by using SPSS 10.0 software. Results 1. The Par - 4 - SiRNA could markedly down - regulate the mRNA levels of Par - 4 gene in hBMSCs. With transfections of Par - 4 - SiRNA, the levels of Par - 4 mRNA were decreased by 88% （ P 〈 0.01 ）. 2. Par - 4 - SiRNA inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were decreased to （39.49 ± 5.01 ） % （ P 〈 0.01 ） from （ 59.12 ± 5.43 ） % （ P 〈 0.01 ）. 3. Western blot assays demonstrated that, glutamate down - regulated the expression of CD2AP proteins in hBMSCs. However, Par -4 - SiRNA could markedly recover the clown - regulation of CD2AP proteins induced by glutamate （ P 〈 0.01 ）. Conclusions SiRNA against Par - 4 gene could inhibit the apoptosis of hBMSCs induced by glutamate. The inhibitory action of Par - 4 expression could markedly recover the down - regulation of CD2AP proteins induced by glutamate.