姜黄素通过Akt-mTOR信号通路调控细胞自噬对骨关节炎大鼠的保护作用研究  

Protective effects of curcumin on chondrocyte in rats with osteoarthritis by regulating autophagy via Akt-mTOR signaling pathway

作　　者：曹舒兴[1] 周楠 王进[3] 赵立辉 曹光[5] 马维[1] 李明[1] CAO Shuxing;ZHOU Nan;WANG Jin(Department of Orthopaedics,The Second Hospital of Hebei Medical University,Hebei,Shijiazhuang 050000,China;不详)

机构地区：[1]河北医科大学第二医院骨科,石家庄市050000 [2]河北医科大学第二医院血液科,石家庄市050000 [3]河北医科大学第二医院药剂科,石家庄市050000 [4]河北省深泽县医院放射科 [5]河北省石家庄市第二医院骨科

出　　处：《河北医药》2021年第24期3685-3689,共5页Hebei Medical Journal

基　　金：河北省医学科学研究重点课题(编号:20190068);河北省财政厅老年病课题(编号:361004)。

摘　　要：目的研究姜黄素对大鼠骨关节炎软骨细胞的保护作用及其与细胞自噬Akt-mTOR信号通路的关系。方法将30只6周龄健康雄性SD大鼠随机分为假手术组、手术组及治疗组。手术组采用Hulth法诱导大鼠骨关节炎模型,治疗组术后即给予口服姜黄素50 mg·kg^(-1)·d^(-1),连续给药8周。通过番红-O染色和Mankin’s评分评估关节软骨结构特征;TUNEL染色分析软骨细胞凋亡情况;Western blot检测凋亡相关标志物caspase-3以及Bax/Bcl2,自噬相关标志物LC-Ⅰ/Ⅱ和Beclin1以及Akt/mTOR信号通路mTOR、Akt、p-P70s6k等。此外,提取进行改良Hulth法手术后的大鼠膝关节软骨组织细胞原代培养,分别添加姜黄素(40μmol/L)、Akt-mTOR信号通路激活剂尼古丁(2.5μmol/L)、姜黄素+尼古丁组(姜黄素处理前2 h给予尼古丁处理),验证姜黄素是否通过Akt-mTOR信号通路调控细胞自噬对骨关节炎大鼠发挥保护作用。结果与手术组相比,姜黄素治疗组Mankin’s评分显著降低(P<0.05);软骨细胞凋亡受到抑制;凋亡相关标志物caspase-3以及Bax/Bcl2比值显著下降(P<0.05);自噬相关标志物LC-Ⅰ/Ⅱ和Beclin1表达显著升高(P<0.05);mTOR、Akt、p-P70s6k的表达水平显著降低(P<0.05)。在细胞实验中,当给予姜黄素+尼古丁处理后,相比于单纯给予姜黄素,LC3Ⅱ/LC3Ⅰ水平明显下降(P<0.01)。结论姜黄素通过Akt/mTOR信号通路诱导自噬,在骨关节炎大鼠中发挥了治疗作用。Objective To investigate the protective effects of curcumin on chondrocyte in rats with osteoarthritis,and to explore ots correlation with Akt-mTOR signaling pathway.Methods Thirty healthy male SD rats aged 6 weeks were randomly divided into sham operation group,operation group and treatment group.The rats models with osteoarthritis were established by Hulth method,then the rats in treatment group were given orally curcumin 50mg·kg^(-1)·d^(-1) immediately after surgery for 8 weeks.The structural characteristics of articular cartilage were assessed by the saffron O staining and Mankin’s score.TUNEL staining was used to analyze the apoptosis of chondrocytes.Western Blot was used to detect apoptosis-related markers-Caspase-3 and Bax/Bcl2,autophagy related markers LC-Ⅰ/Ⅱand Beclin1,and Akt/mTOR signaling pathway mTOR,Akt and p-p70S6K.In addition,the rat knee cartilage tissue cells primary culture was performed after the operation,which were treated by curcumin(40μmol/L),Akt-mTOR signal pathway activator nicotine(2.5μmol/L),or both of them to verify whether curcumin could regulate autophagy through Akt-mTOR signaling pathway to protect the rats with osteoarthritis.Results As compareed with those in operation group,the Mankin’s scores were significantly decreased in treated group(P<0.05),and the apoptosis of chondrocytes was inhibited,and the ratio of Caspase-3 and Bax/Bcl2 ratio was significantly decreased(P<0.05),however,the expression levels of autophagy-related markers LC-Ⅰ/Ⅱand Beclin1 were significantly increased(P<0.05),but the expression levels of mTOR,Akt and p-p70S6K were significantly decreased(P<0.05).In the cell experiment,the levels of LC3Ⅱ/LC3Ⅰwere decreased significantly after the celle were treated by curcumin combined with nicotine,as compared with those by simple curcumin(P<0.01).Conclusion Curcumin can induce autophagy through the Akt/mTOR signaling pathway and play a therapeutic role in rats with osteoarthritis.

关 键 词：姜黄素 骨关节炎 自噬作用 Akt-mTOR信号通路 

分 类 号：R684.3[医药卫生—骨科学]