姜黄素抑制硝普钠诱导的大鼠软骨细胞凋亡机制研究  

Curcumin inhibits sodium nitroprusside-induced apoptosis of rat chondrocytes via mitochondria-dependent pathway

作　　者：袁伟[1] 卞阳阳 朱恒杰 程少文[1] 彭磊[1] YUAN Wei;BIAN Yang-yang;ZHU Heng-Jie(Trauma Center,the First Affiliated Hospital of Hainan Medical University,Haikou,Hainan 570102,China)

机构地区：[1]海南医学院第一附属医院创伤医学中心,海南海口570102

出　　处：《中华全科医学》2019年第4期528-532,共5页Chinese Journal of General Practice

基　　金：国家自然科学基金项目(81460339)

摘　　要：目的探讨姜黄素抑制硝普钠(SNP)诱导的大鼠软骨细胞凋亡的机制研究。方法每组取4只SD大鼠关节软骨,细胞进行原代培养,将软骨细胞分成6组,分别为空白对照组(A组),SNP损伤组(B组),SNP+5μmol/L姜黄素组(C组),SNP+10μmol/L姜黄素组(D组),SNP+20μmol/L姜黄素组(E组),SNP+30μmol/L姜黄素组(F组)。用CCK8法检测不同浓度的姜黄素对软骨细胞增殖的影响,同时检测各组中软骨细胞的活性;用RtPCR检测软骨细胞MMP-13、caspase-3、Bax和Bcl-2基因的表达情况;用Hoechst 33342染色观察软骨细胞核凋亡情况;Western Blotting检测线粒体凋亡途径相关蛋白的表达情况。结果①当姜黄素的浓度为20μmol/L时,姜黄素对软骨细胞的促增殖作用最为明显;②CCK8法检测各组细胞的活性率从大至小依次为A组>E组>D组>C组>F组>B组;③RtPCR检测软骨细胞caspase-3、Bax和Bcl-2表达量经姜黄素处理后均有显著变化;④Hoechst 33342染色结果表明姜黄素能改善SNP诱导的软骨细胞核染色质的损伤,减少软骨细胞中的线粒体膜电位;⑤Western boltting检测各组软骨细胞中caspase-3、Bax、蛋白的表达情况,表达量依次为B组>E组>A组;Bcl-2蛋白的表达情况,表达量依次为A组>E组>B组。结论姜黄素通过抑制线粒体凋亡途径,从而减少SNP诱导的大鼠软骨细胞的凋亡。Objective To research the mechanism that curcumin inhibits sodium nitroprusside(SNP)-induced apoptosis of chondrocytes in rats. Methods The carilage cells were obtained from articular cartilages of SD rats(4 rats in each group). The cartilage cells were divided into control group(A), the SNP injured group(B), SNP+5 μmol/L curcumin group(C), SNP+10 μmol/L curcumin group(D), SNP+ 20 μmol/L curcumin group(E), and SNP+ 30 μmol/L curcumin group(F). The effects of different concentrations of curcumin on the proliferation of chondrocytes and the activity of chondrocytes in each group were detected by CCK8 method. The expression of MMP-13, caspase-3, Bax and Bcl-2 genes in chondrocytes were detected by RtPCR. The nuclear apoptosis of chondrocytes were detected by Hoechst 33342 staining. Western blotting technique was used to detect the expression of mitochondrial apoptosis pathway-associated protein. Results ① The 20 μmol/L curcumin had the most proliferative effect on chondrocytes.② The cells activity by CCK8 assay: group A>group E> group D>group C>group F>group B.③In group C, D, E and F, the expression of caspase-3, Bax and Bcl-2 in chondrocytes was significantly different from group A and B.④Hoechst 33342 staining showed that curcumin inhibit SNP-induced chondrocyte nuclear chromatin damage and reduce mitochondrial membrane potential in chondrocytes.⑤The caspase-3 and Bax protein expression of chondrocyte by Western blotting: group B>group E>group A;The Bcl-2 protein expression of chondrocyte by Western blotting: group A>group E>group B. Conclusion The curcumin can prevent SNP-induced rat chondrocyte apoptosis via inhibition of mitochondria-dependent pathway.

关 键 词：姜黄素 骨关节炎 软骨细胞 线粒体凋亡 

分 类 号：R681.53[医药卫生—骨科学] R329.25[医药卫生—外科学]