羟基红花黄素A拮抗IL-1β诱导软骨终板细胞凋亡的作用机制  

The anti-apoptotic effect of Hydroxy safflower yellow A on end-plate chondrocytes induced by IL-1β

作　　者：张宇[1,2] 赵永见[3,2] 周泉[3,2] 王拥军[3,2] 郑为超[1] 

机构地区：[1]安徽理工大学医学院,安徽淮南232001 [2]上海中医药大学脊柱病研究所,上海200032 [3]上海中医药大学附属龙华医院,上海200032

出　　处：《中国药理学通报》2011年第12期1704-1709,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30901914,30801478,30701118);国家教育部高等学校博士点基金资助项目(No 20093107110003);上海市优秀学科带头人计划资助项目(No 08XD1404000);上海市高校创新团队计划(沪教委科[2009]6号);安徽省自然科学基金项目(No11040606M203);安徽高校省级自然科学研究重点项目(No KJ2009A038)

摘　　要：目的探讨羟基红花黄素A(hydroxy safflower yellowA,HSYA)对IL-1β诱导小鼠软骨终板细胞凋亡的拮抗作用。方法用10μg.L-1的IL-1β诱导正常软骨终板细胞退变,分别制备不同浓度的HSYA培养基作为干预措施,采用流式细胞仪检测细胞凋亡,RT-PCR检测凋亡相关基因的表达水平。采用Western blot检测细胞内Bax、Bcl-2蛋白表达量,细胞免疫荧光分析各组Bax、Bcl-2细胞内定位。结果不同浓度的HSYA培养基均能拮抗IL-1β诱导的软骨细胞凋亡。流式细胞分析结果显示各浓度HSYA可以降低IL-1β诱导的小鼠椎间盘软骨终板细胞的凋亡比例;RT-PCR结果显示各浓度HSYA可拮抗IL-1β诱导细胞上调Bax基因表达,下调Bcl-2基因表达作用;细胞免疫荧光结果显示HSYA 10-5 mol.L-1给药组可下调Bax表达,上调Bcl-2表达;Western blot结果显示HSYA 10-5 mol.L-1和HSYA10-6 mol.L-1均可以拮抗IL-1β诱导细胞上调Bax蛋白表达,下调Bcl-2蛋白表达作用。结论羟基红花黄素A具有拮抗IL-1β诱导小鼠椎间盘软骨终板细胞凋亡的作用。Aim To observe the anti-apoptotic effect of Hydroxy safflower yellow A on IL-1β induced end-plate chondrocytes in mice.Methods The cell was induced by IL-1β（10 μg·L-1）and the apoptosis was cured by HSYA with different concentrations.Then the effect on apoptosis by FCM and RT-real time PCR was tested.The level of Bax and Bcl-2 protein expression was detected by Western blot and immunofluorescence.Results The apoptosis induced by IL-1β could be inhibited by HSYA with different concentrations.HSYA could reduce the apoptotic rate analysed by FCM,down-regulate the gene expression of Bax and up-regulate Bcl-2 assayed by RT-real time PCR.The result approved by immunofluorescence was that HSYA 10-5mol·L-1could up-regulate the protein expression of Bax and down-regulate the expression level of Bcl-2.Besides,observation of the protein expression of Bax and Bcl-2 by Western blot found HSYA 10-5mol·L-1 and HSYA 10-6mol·L-1could inhibit the effect of IL-1β,which could increase the level of protein expression of Bax and reduce the expression level of Bcl-2.Conclusion HSYA has an effect on anti-apoptosis of end-plate chondrocytes in mice induced by IL-1β.

关 键 词：羟基红花黄素A 软骨终板 凋亡 IL-1Β HSYA 软 骨细胞 

分 类 号：R-332[医药卫生] R284.1