摘要
目的明确调节细胞内NAD水平对体外髓核细胞退行性变指标的影响。方法分组利用FK866及NMN处理大鼠尾椎椎间盘髓核细胞。CCK-8法检测髓核细胞的增殖的影响。WST-8法检测NAD含量;RT-PCR测定退变相关指标的表达水平。结果FK866处理下的髓核细胞NAD含量减少,退行性变水平增加(P<0.05);NMN处理后的髓核细胞NAD含量增加,但退行性变水平改变差异无统计学意义(P>0.05);NMN干预FK866预处理的髓核细胞,NAD含量升高,较低浓度可恢复降低髓核细胞的退行性变水平(P<0.05),但高浓度处理可加重退行性变(P<0.05)。结论NAD的含量显著降低与髓核细胞的退行性变相关,但补充需要维持在合理剂量,NAD缺失的髓核细胞施用大量NMN可能加重退行性变;NAD池的稳态对于维持髓核细胞正常功能有重要意义。
Objective To demonstrate the effect of regulating intracellular NAD levels on the degeneration indexes of nucleus pulposus cells in vitro.Methods FK866 and nicotinamide mononucleotide was used to treat nucleus pulposus cells.CCK-8 assay was used to detect the effects on the proliferation of cells.WST-8 method was used to detect NAD content.RT-PCR experiment was used to determine the expression level of degenerationrelated genes.Results The NAD content of nucleus pulposus cells under FK866 treatment decreased,and the degeneration level increased(P<0.05).As for NMN treatment,the NAD content increased,but there was no statistically significant change in degeneration(P>0.05).After NMN treatment of nucleus pulposus cells following FK866 treatment,NAD content revived,lower concentration treatment restores the expression level of Col-2(P<0.05),but high concentration treatment can aggravate the degeneration level(P<0.05).Conclusion Significant reduction in NAD content relates to degeneration of nucleus pulposus cells,but supplementation needs to be maintained at a reasonable dose:administration of large amounts of NMN to nucleus pulposus cells lacking NAD may aggravate degeneration.The homeostasis of the NAD pool is important for maintaining the normal function of nucleus pulposus cells.
作者
许琮欣
堵嘉诚
杜彤
谢依彤
王鹏博
戴顺捷
毛海青
杨惠林
XU Cong-xin;DU Jia-cheng;DU Tong;XIE Yi-tong;WANG Peng-bo;DAI Shun-jie;MAO Hai-qing;YANG Hui-lin(Department of Orthopedics,the First Affiliated Hospital of Soochow University,Suzhou,Jiangsu,215006,China;Medical College of Soochow University,Suzhou,Jiangsu,215123,China)
出处
《中国血液流变学杂志》
CAS
2021年第2期158-162,共5页
Chinese Journal of Hemorheology
基金
国家自然科学基金资助项目(91849114)。