精脒通过激活自噬促进衰老兔BMSCs骨向分化的实验研究  

Spermidine promotes osteogenic differentiation of senescent rabbit bone-marrow mesenchymal stem cells via activating autophagy

作　　者：马雨聪 杨文朋 吴方丽 田萧羽 李琳[4] 李爽 冯晋[4] 朱彪 MA Yu-cong;YANG Wen-peng;WU Fang-li;TIAN Xiao-yu;LI Lin;LI Shuang;FENG Jin;ZHU Biao(Department of Stomatology,Beijing Sixth Hospital,Beijing 100007,China;Department of Stomatology,Aero Space Center Hospital,Beijing 100049,China;Department of Stomatology,Rocket Force Characteristic Medical Center,Beijing 100088,China;Department of Stomatology,the First Medical Center,Chinese People’s Liberation Army General Hospital,Beijing 100853,China;Department of Stomatology,Fuxing Hospital,Capital Medical University,Beijing 100038,China)

机构地区：[1]北京市第六医院口腔科,北京100007 [2]航天中心医院口腔科,北京100049 [3]火箭军特色医学中心口腔科,北京100088 [4]解放军总医院第一医学中心口腔科,北京100853 [5]首都医科大学附属复兴医院口腔科,北京100038

出　　处：《临床口腔医学杂志》2021年第12期707-711,共5页Journal of Clinical Stomatology

基　　金：中国博士后科学基金资助项目(2019T120980)。

摘　　要：目的:探讨精脒(Spermidine)对衰老兔骨髓间充质干细胞(BMSCs)骨向分化的影响及其自噬机制。方法:复苏兔BMSCs并将其分为4组:对照组(Vehicle),D-半乳糖诱导衰老组(D-gal),Spermidine组(Spermidine)和自噬抑制剂氯奎干预组(CQ)。MTT法检测细胞增殖。成骨诱导后,qPCR检测成骨基因Runx2、Osx、ALP和OCN的转录表达;Western Blot检测成骨蛋白ALP,自噬蛋白P62和LC3B的表达。结果:与Vehicle组相比,D-gal明显抑制BMSCs增殖(P<0.05);而Spermidine可促进BMSCs增殖(P<0.05)。成骨诱导培养后,D-gal下调成骨基因Runx2、Osx和ALP的转录表达,抑制ALP蛋白表达,提高P62蛋白表达并降低LC3B-II蛋白表达(P<0.05)。而Spermidine可上调Runx2、Osx和ALP的转录表达,促进ALP蛋白表达,降低P62蛋白表达并提高LC3B-II蛋白表达水平(P<0.05)。当采用氯奎抑制自噬后,Spermidine上调成骨基因转录表达和促进ALP蛋白表达的作用明显减弱(P<0.05)。结论:Spermidine通过激活自噬促进衰老兔BMSCs骨向分化,为临床上缓解老龄性骨质疏松并进而提高口腔种植手术成功率提供潜在的新药物。Objective:To investigate the effects of Spermidine on osteogenic differentiation of senescent rabbit BMSCs and autophagy-related mechanisms.Methods:BMSCs were thawed and divided into 4 groups:Vehicle,D-gal(D-galactose),Spermidine(D-gal plus Spermidine administration)and CQ(chloroquine;Spermidine plus chloroquine intervention).Cell viability was detected using MTT method.After cultured in osteogenic medium,mRNA levels of Runx2,Osx,ALP and OCN and protein levels of ALP,P62 and LC3B were investigated.Results:Compared to the Vehicle group,D-gal significantly inhibited BMSCs proliferation,while spermidine promoted BMSCs proliferation(P<0.05).After cultured in osteogenic medium,Runx2,Osx and ALP mRNA expressions were downregulated,ALP protein level was decreased.P62 protein level was increased and LC3B-Ⅱprotein level was decreased by D-gal administration(P<0.05).However,spermidine promoted mRNA expressions of Runx2,Osx and ALP,increased ALP protein level,decreased P62 protein level and increased LC3B-Ⅱprotein level(P<0.05).When suppressed autophagy using CQ,the effects of spermidine on up-regulating osteogenic genes and increasing ALP protein levels were blunted(P<0.05).Conclusion:Spermidine could promote osteogenic differentiation of senescent rabbit BMSCs via activating autophagy,which could be considered as a potential strategy for attenuating senile osteoporosis and further contributing to successful operation of oral implantation.

关 键 词：精脒 D-半乳糖 衰老 自噬 骨向分化 

分 类 号：R780.2[医药卫生—口腔医学]