摘要
为探索二苯乙烯苷(THSG)保护成骨细胞抗氧化损伤的作用机制,本试验使用不同浓度THSG预处理MC3T3-E1成骨样细胞1 h,加入0.3 mmol/L过氧化氢(H_(2)O_(2))共同培养细胞,采用噻唑蓝(MTT)法检测细胞生存活力;实时荧光定量聚合酶链式反应(FQ-PCR)法检测细胞Bcl-xL/Bcl-2相关死亡启动因子(Bad)、骨保护素(OPG)、核因子κB受体活化因子配体(RANKL)和β-连环蛋白(β-catenin)mRNA表达水平;Western blot法检测细胞OPG和β-catenin蛋白水平。结果显示,与对照组相比,H_(2)O_(2)组细胞生存活力显著下降(P<0.05),Bad和RANKL mRNA表达水平显著上升(P<0.05),OPG和β-catenin mRNA和蛋白水平均显著下降(P<0.05);与H_(2)O_(2)组相比,THSG(10^(-4)mg/mL)保护组细胞生存活力显著上升(P<0.05),Bad和RANKL mRNA表达水平显著下降(P<0.05),OPG和β-catenin mRNA和蛋白水平均显著上升(P<0.05)。结果表明,THSG能够拮抗H_(2)O_(2)对成骨细胞生存活力、OPG/RANKL mRNA比和Wnt/β-catenin通路的抑制作用,保护成骨细胞抗氧化损伤。
This study aimed to explore the protective mechanism of tetrahydroxystilbene glucoside(THSG)against oxidative damage in osteoblast.Firstly,osteoblastic MC3T3-E1 cells were treated with 0.3 mmol/L hydrogen peroxide(H_(2)O_(2))after THSG pre-treatment at different concentrations for 1 h,then,MTT assays were used to assess the cell viability;fluorescence quantitative PCR(FQ-PCR)was used to determine the mRNA levels of Bcl-XL/Bcl-2 associated death promoter(Bad),osteoprotectin(OPG),receptor activator for nuclear factor kappa B ligand(RANKL)andβ-catenin;Western blot was used to detect the protein levels of OPG andβ-catenin.The results showed that,compared with the control group,H_(2)O_(2)significantly decreased the cell viability(P<0.05),significantly increased the mRNA levels of Bad and RANKL(P<0.05),and significantly decreased the mRNA and protein levels of OPG andβ-catenin(P<0.05)in MC3T3-E1 cells;compared with the H_(2)O_(2)group,the THSG(10^(-4) mg/mL)pre-treatment significantly increased the cell viability(P<0.05),significantly decreased the mRNA levels of Bad and RANKL(P<0.05),and significantly increased the mRNA and protein levels of OPG andβ-catenin(P<0.05)in MC3T3-E1 cells.Thus,THSG could antagonize the inhibitory effect of H_(2)O_(2)on the viability,OPG/RANKL mRNA ratio and Wnt/β-catenin pathway in osteoblasts,and protect osteoblasts from oxidative damage.
作者
范迎赛
宫新城
杨倩
张迪
赵兴华
王晓丹
史万玉
FAN Ying-sai;GONG Xin-cheng;YANG Qian;ZHANG Di;ZHAO Xing-hua;WANG Xiao-dan;SHI Wan-yu(College of Traditional Chinese Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China)
出处
《中国兽医杂志》
CAS
北大核心
2022年第4期6-11,共6页
Chinese Journal of Veterinary Medicine
基金
河北省高等学校科学技术研究项目(QN2020269)。
