6-gingerol protects nucleus pulposus-derived mesenchymal stem cells from oxidative injury by activating autophagy  

作　　者：Li-Ping Nan Feng Wang Yang Liu Zhong Wu Xin-Min Feng Jun-Jian Liu Liang Zhang 

机构地区：[1]Department of Orthopedic,Tongji University School of Medicine,Shanghai Tenth People’s Hospital,Tenth People’s Hospital of Tongji University,Shanghai 200072,China [2]Department of Spine Surgery,Tongji University School of Medicine,Shanghai East Hospital,Shanghai 200120,China [3]Department of Orthopedic,West China Hospital of Sichuan University,Chengdu 610000,Sichuan Province,China [4]Department of Orthopedics,Shanghai Tenth People’s Hospital,Tongji University School of Medicine,Shanghai 200072,China [5]Department of Orthopedics,Clinical Medical College of Yangzhou University,Yangzhou 225000,Jiangsu Province,China

出　　处：《World Journal of Stem Cells》2020年第12期1603-1622,共20页世界干细胞杂志（英文版）（电子版）

基　　金：Supported by National Natural Science Foundation of China,No.81972136;National Natural Science Foundation for Young Scholars of China,No.81401830;Guangxi Natural Science Foundation General Project,No.2018JJA14775;Young Medical Scholars Major Program of Jiangsu Province,No.QNRC2016342;Innovation Team Project of Jiangsu Province,No.CXTDB2017004;and Key Funding Project of Maternal and Child Health Research of Jiangsu Province,No.F201801.

摘　　要：BACKGROUND To date,there has been no effective treatment for intervertebral disc degeneration(IDD).Nucleus pulposus-derived mesenchymal stem cells(NPMSCs)showed encouraging results in IDD treatment,but the overexpression of reactive oxygen species(ROS)impaired the endogenous repair abilities of NPMSCs.6-gingerol(6-GIN)is an antioxidant and anti-inflammatory reagent that might protect NPMSCs from injury.AIM To investigate the effect of 6-GIN on NPMSCs under oxidative conditions and the potential mechanism.METHODS The cholecystokinin-8 assay was used to evaluate the cytotoxicity of hydrogen peroxide and the protective effects of 6-GIN.ROS levels were measured by 2´7´-dichlorofluorescin diacetate analysis.Matrix metalloproteinase(MMP)was detected by the tetraethylbenzimidazolylcarbocyanine iodide assay.TUNEL assay and Annexin V/PI double-staining were used to determine the apoptosis rate.Additionally,autophagy-related proteins(Beclin-1,LC-3,and p62),apoptosisassociated proteins(Bcl-2,Bax,and caspase-3),and PI3K/Akt signaling pathwayrelated proteins(PI3K and Akt)were evaluated by Western blot analysis.Autophagosomes were detected by transmission electron microscopy in NPMSCs.LC-3 was also detected by immunofluorescence.The mRNA expression of collagen II and aggrecan was evaluated by real-time polymerase chain reaction(RT-PCR),and the changes in collagen II and MMP-13 expression were verified through an immunofluorescence assay.RESULTS 6-GIN exhibited protective effects against hydrogen peroxide-induced injury in NPMSCs,decreased hydrogen peroxide-induced intracellular ROS levels,and inhibited cell apoptosis.6-GIN could increase Bcl-2 expression and decrease Bax and caspase-3 expression.The MMP,Annexin V-FITC/PI flow cytometry and TUNEL assay results further confirmed that 6-GIN treatment significantly inhibited NPMSC apoptosis induced by hydrogen peroxide.6-GIN treatment promoted extracellular matrix(ECM)expression by reducing the oxidative stress injury-induced increase in MMP-13 expression.6-GIN activated autopha

关 键 词：Nucleus pulposus-derived mesenchymal stem cells 6-GINGEROL Intervertebral disc degeneration Oxidative stress AUTOPHAGY Apoptosis 

分 类 号：R285[医药卫生—中药学]