中国临床药理学杂志2024，Vol.40Issue(3)：373-377,5.DOI:10.13699/j.cnki.1001-6821.2024.03.015
1,25(OH)2D3调控维生素D受体表达对H2O2诱导的髓核细胞氧化应激损伤的影响
Effects of 1,25(OH)2 D3 regulation of vitamin D receptor expression on H2 O2-induced oxidative stress injury in nucleus pulposus cells
摘要
Abstract
Objective To investigate the effect of 1,25(OH)2 D3-mediated vitamin D receptor(VDR)expression on oxidative stress injury in nucleus pulposus cells.Methods Human interdisc nucleus pulpocytes were randomly divided into control group,H2O2 group(400 µmol·L-1H2O2),1,25(OH)2 D3 group[100.00 nmol·L-11,25(OH)2D3+400 μmol·L-1H2O2],1,25(OH)2D3+si-NC group[100.00 nmol·L-1 1,25(OH)2D3+400 μmol·L-1 H2O2+si-NC],1,25(OH)2D3+si-VDR group[100.00 nmol·L-1 1,25(OH)2D3+400 μmol·L-1 H2O2+si-VDR].Cell survival rate was measured by methyl thiazolyl tetrazolium(MTT)assay.Western blot assay was used to detect the protein expression level of each group.Apoptosis,mitochondrial membrane potential and reactive oxygen species(ROS)levels were detected by flow cytometry,JC-1 fluorescence probe and DCFH-DA probe,respectively.Malonclialdehyde(MDA)and superoxide dismutase(SOD)levels were detected with the kit.Results The cell survival rates of control group,H2O2 group,1,25(OH)2D3 group,1,25(OH)2D3+si-NC group and 1,25(OH)2D3+si-VDR group were(100.00±2.43)％,(63.79±5.21)％,(90.11±7.24)％,(88.79±6.48)％and(55.31±4.65)％;VDR protein levels were 0.79±0.06,0.28±0.03,0.68±0.05,0.69±0.06 and 0.34±0.04;the apoptosis rates were(4.12±0.26)％,(19.37±1.21)％,(8.49±0.57)％,(8.23±0.60)％and(14.68±1.37)％;mitochondrial membrane potential levels were(100.00±4.31)％,(49.46±5.02)％,(82.14±7.02)％,(83.14±5.12)％and(67.16±5.48)％;ROS levels were 1.79±0.12,7.98±0.51,3.87±0.34,3.92±0.22 and 5.79±0.28;Beclin-1 levels were 0.86±0.09,0.35±0.04,0.76±0.07,0.75±0.08 and 0.46±0.05;LC3-Ⅱ/LC3Ⅰ levels were 1.00±0.07,0.25±0.04,0.78±0.07,0.85±0.08 and 0.42±0.05,respectively.Compared H2 O2 group with control group,compared 1,25(OH)2 D3 group with H2O2 group,compared 1,25(OH)2D3+si-VDR group with 1,25(OH)2D3+si-NC group,the differences of the above indicators were all statistically significant(all P＜0.05).Conclusion 1,25(OH)2 D3 may up-regulate the expression of VDR,promote autophagy and inhibit apoptosis of nucleus pulposus cells,and play an anti-oxidative stress role.关键词
1,25(OH)2D3/髓核细胞/维生素D受体/氧化应激/自噬Key words
1,25(OH)2D3/nucleus pulposus cells/vitamin D receptor/oxidative stress/autophagy分类
医药卫生引用本文复制引用
沈哲,蓝涛,郭伟壮..1,25(OH)2D3调控维生素D受体表达对H2O2诱导的髓核细胞氧化应激损伤的影响[J].中国临床药理学杂志,2024,40(3):373-377,5.基金项目
广东省卫生健康委医学科研基金资助项目(B2022061) （B2022061）
