邻苯二甲酸酯对软骨细胞凋亡及超微结构的影响  

Impact of phthalic acid esters on the apoptosis and ultrastructure modification of chondrocytes

作　　者：张银昌[1] 王强[1] 宣华兵[1] 贺华正[1] 杨昕[1] 

机构地区：[1]皖南医学院附属弋矶山医院关节骨科,安徽芜湖241001

出　　处：《皖南医学院学报》2015年第2期110-113,共4页Journal of Wannan Medical College

基　　金：皖南医学院中青年科研基金项目(WK2012F07)

摘　　要：目的:通过研究塑化剂邻苯二甲酸酯对软骨细胞凋亡因子bcl-2和Bax蛋白表达及超微结构的影响,探讨邻苯二甲酸酯在致软骨细胞凋亡中的作用机制。方法:用软骨细胞体外培养的方法,原代培养乳鼠关节软骨细胞,传第3代后加药,按染邻苯二甲酸酯剂量不同分为0/L(对照)、10-10/L、10-8/L、10-6/L组,染邻苯二甲酸酯培养7 d后,透射电镜下观察软骨细胞的超微结构改变,采用western-blot方法检测各实验组中软骨细胞bcl-2和Bax蛋白的表达。结果:透射电镜下对照组和10-10/L组的软骨细胞呈球形,粗面内质网发达,线粒体膜性结构完整;10-8/L、10-6/L组的细胞内可见脂滴明显增多,胞质内出现大量空泡类物质,细胞内膜结构不清,部分细胞出现核固缩。10-8/L、10-6/L组的软骨细胞bcl-2的蛋白表达(0.620±0.041和0.530±0.035)较对照组表达(0.805±0.032)显著降低(P<0.01);而Bax的表达(0.966±0.038和1.280±0.154)较对照组表达(0.602±0.045)显著增加(P<0.01)。10-10/L组软骨细胞中bcl-2的表达(0.878±0.055)和Bax的表达(0.650±0.039)与对照组相比均无统计学差异(P>0.05)。此外,10-8/L、10-6/L组相比,bcl-2和Bax的表达也有显著差异(P<0.01)。结论:10-8/L、10-6/L浓度的邻苯二甲酸酯可对软骨细胞超微结构造成损伤,其通过减少抑凋亡因子bcl-2的表达和增加促凋亡因子Bax的表达,从而产生促进软骨细胞凋亡的作用。Objective：To in vitro investigate the effects phthalie acid esters（ PAEs, a pastieizer） on the expression of Bcl-2 and Bax in ehondrocytes and ultrastructure modification as well as the mechanisms in generating apoptosis of the ehondrocytes. Methods ： Primary articular chondrocytes of neonate rat were cultured with in vitro technique by the third generation. Then the ehrondrocytes were infected with PAEs by concentration of 0/L, 10- ^-10/L, 10^-8/L and 10^-6/L, respectively, for 7 days, and observed under transmission electron microscope to examine the changes of their ultrastrncture. Western blot was performed to determine the expression of Bcl-2 and Bax in chondrocytes in different dosage groups. Results： Microscopically, the chrondroeytes in dosage group of 10 ^-10/L were in round shape, with abundant rough endoplasmic retieulum （RERs） and complete structure of mitochondria membrane, while in dosage groups of 10^-8/L and 10^-6/L, displayed intensified intracellular lipid droplets, with a number of vacuoles being seen in the cytoplasm, incomplete structure of the intracellular membranes and partial pyknosis of the ehromatine. Bcl-2 expression was significantly down-regulated in dosage group of 10 ^-8/ L（ O. 620 ± 0. 041 ） and 10^-6/L（0. 530 ± 0. 035 ） compared to the control group （0. 805 ± 0. 032 ） （ P 〈 0.01 ）, whereas Bax significantly increased （ 0. 966 ± 0. 038 for 10^-8/L, and 1. 280 ± 0. 154 for 10 -6/L） compared to the control group （ Bcl-2 ： 0. 602 ± 0. 045 ; Bax： 0. 602 ± 0. 045 ） （ P 〈 0.01 ）. There was no statistical difference regarding the expression of Bcl-2 or Bax between 10^-10/L group and control group（P 〉0.05 ） ,yet the difference was significant by expression of Bcl-2 and Bax in dosage group of 10^-8/L and 10^-6/L（ P 〈 0.01 ）. Conclusion ： PAE at concentration of either 10^-8/L or 10^-6/L may lead to damage to the ultrastructure of chondrocyte. The potential mechanism may be involved in inactivation of the anti-apoptotie

关 键 词：邻苯二甲酸酯 软骨细胞 BCL-2 BAX 

分 类 号：R684[医药卫生—骨科学]