姜黄素通过上调PPARγ抑制糖基化终末产物诱导的软骨细胞凋亡及线粒体功能损伤  

Curcumin inhibits advanced glycation end products-induced cell apoptosis and mitochondrial dysfunction via elevating PPARγ in chondrocyte

作　　者：杨青山[1] 吴树金[2] 施松波[1] 毛新展[3] 郭士方[1] 陈志信[1] 台会平[1] 

机构地区：[1]甘肃省人民医院骨科,甘肃兰州730000 [2]甘肃省人民医院药剂科,甘肃兰州730000 [3]中南大学湘雅二医院骨科,湖南长沙410011

出　　处：《中国药理学通报》2018年第2期261-267,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81760409);甘肃省卫生行业科研计划资助项目(No GSWSKY-2014-10);甘肃省中医药管理局科研计划项目(No GZK-2013-42)

摘　　要：目的观察姜黄素(curcumin)对糖基化终末产物(AGEs)诱导的软骨细胞凋亡及线粒体功能损伤的作用,并探讨相关机制是否与姜黄素上调过氧化物酶体增殖物激活受体-γ(PPARγ)相关。方法原代培养家兔膝关节软骨细胞;TUNEL染色法检测细胞凋亡;Rhodamine-123试剂盒检测线粒体跨膜电位(△Ψm);ATP试剂盒检测线粒体ATP生成;分光光度法检测caspase-3活性;Western blot检测PPARγ、细胞色素C、Bax及Bcl-2蛋白表达;real-time PCR检测PPARγmRNA含量;DNA-binding法检测PPARγ活性。结果 AGEs(200 mg·L-1)可明显诱导兔软骨细胞凋亡,同时线粒体跨膜电位(△Ψm)降低、ATP生成减少,caspase-3活性增加,细胞色素C释放增加,Bax/Bcl-2的比值增加;线粒体通透性转换孔的抑制剂环孢霉素A(Cs A,100 nmol·L-1)可以明显抑制AGEs诱导的细胞凋亡;PPARγ特异性激动剂吡格列酮及姜黄素均可明显抑制AGEs诱导的软骨细胞凋亡及线粒体功能损伤,给予PPARγ特异性抑制剂GW9662 10μmol·L-1预处理后,可以明显拮抗姜黄素的保护作用。同时,姜黄素可以明显上调AGEs诱导的PPARγ活性的降低,并伴随PPARγ相应的mRNA及蛋白表达水平的升高。结论姜黄素通过上调PPARγ,有效地保护AGEs诱导的软骨细胞线粒体损伤,从而抑制AGEs诱导的软骨细胞凋亡。Aim To explore the mechanism of the protective effect of curcumin on advanced glycation end products( AGEs)-induced chondrocyte apoptosis and mitochondrial dysfunction whether by elevating peroxisome proliferators-activated receptor-γ( PPARγ) or not. Methods The ratio of apoptotic cells was assayed by TUNEL; the mitochondrial membrane potential( △Ψm) was evaluated by Rhodamine-123 fluorescence. The ATP content was assayed by related kits.The activity of caspase-3 was detected by spectrophotometry. The expression of cytochrome C,Bax,and Bcl-2 was detected by Western blot. The PPARγ expression was determined by Western blot and real-time PCR; in addition,its activity was assayed by DNAbinding method. Results AGEs could induce chondrocyte apoptosis and up-regulate the levels of cytochrome C and caspase-3. Simultaneously,AGEs decreased the levels of △Ψm and ATP production. Mitochondrial permeability conversion pore inhibitor cyclosporine A could significantly protect the cells from apoptosis. In addition,both PPARγ specific agonist pioglitazone and curcumin significantly inhibited AGEs-induced chondrocytes apoptosis and mitochondrial dysfunction. However,pretreatment with PPARγ specific inhibitor GW9662( 10 μmol·L-1) could significantly antagonize the protective effect of curcumin on mitochondrial damage induced by AGEs. Curcumin could also significantly increase PPARγ transcriptional activity induced by AGEs,together with a significant induction of PPARγ protein and mRNA expression. Conclusion Curcumin could effectively protect AGEs-induced chondrocyte mitochondrial dysfunction by upregulating PPARγ,thus protecting chondrocytes from apoptosis.

关 键 词：糖基化终末产物 线粒体损伤 过氧化物酶体增殖物激活受体-Γ 姜黄素 凋亡 软骨细胞 

分 类 号：R-332[医药卫生] R282.71