Department of Spine Surgery, The Third Hospital of HeBei Medical University, 139 Ziqiang Road, Shijiazhuang, 050051, China.
J Orthop Surg Res. 2021 Jun 28;16(1):411. doi: 10.1186/s13018-021-02545-9.
To observe the effects of vitamin D on the apoptotic human nucleus pulposus cells under tumor necrosis factor-α (TNF-α) treatment.
The gene expression data was downloaded from the NCBI Gene Expression Omnibus (GEO) database ( https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE34095 ). Differentially expressed genes between degenerative disc and non-degenerative disc were performed by R software. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analyses were performed using The Database for Annotation, Visualization and Integrated Discovery (DAVID). Then, the human nucleus pulposus tissue was harvested from 12 patients according to the modified Pfirrmann classification and human nucleus pulposus cells were obtained from digestion of herniated nucleus pulposus tissue. The collected nucleus pulposus cells were treated with different concentration of TNF-α, and cellular apoptosis was measured by flow cytometry. Then, human nucleus pulposus cells were divided into following groups: normal culture medium, TNF-α treated, TNF-α, and vitamin D-treated groups. Cellular apoptosis rate was quantified by flow cytometry. Protein expression of p-p65, p65, and IkBa was detected with western blot analysis.
A total of 536 differentially expressed genes were identified through bioinformatic analysis. KEGG pathway revealed that NF-kB signaling pathway was involved in the process of disc degeneration. In the NP cell cultures, vitamin D significantly increased cell proliferation potency. Furthermore, vitamin D inhibited TNF-α induced apoptosis of human nucleus pulposus cells. Vitamin D reduced the phospho-NF-κB/p65 expression in the TNF-α-treated NP cells.
Vitamin D can attenuate TNF-α-induced NP cells apoptosis through interfering with the NF-κB pathway.
观察维生素 D 在肿瘤坏死因子-α（TNF-α）作用下对人髓核细胞凋亡的影响。
从 NCBI 基因表达综合数据库（GEO）（https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE34095）下载基因表达数据。使用 R 软件分析退变和非退变椎间盘之间的差异表达基因。使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路富集分析（DAVID）进行 GO 和 KEGG 通路富集分析。然后，根据改良的 Pfirrmann 分级法从 12 例患者中采集人髓核组织，并从疝出的髓核组织消化中获得人髓核细胞。用不同浓度的 TNF-α处理收集的髓核细胞，用流式细胞术检测细胞凋亡。然后，将人髓核细胞分为以下几组：正常培养基组、TNF-α 处理组、TNF-α+维生素 D 处理组。用流式细胞术定量细胞凋亡率。用 Western blot 分析检测 p-p65、p65 和 IkBa 的蛋白表达。
通过生物信息学分析共鉴定出 536 个差异表达基因。KEGG 通路显示 NF-kB 信号通路参与椎间盘退变过程。在 NP 细胞培养中，维生素 D 显著增加细胞增殖能力。此外，维生素 D 抑制 TNF-α诱导的人髓核细胞凋亡。维生素 D 降低了 TNF-α处理的 NP 细胞中磷酸化 NF-κB/p65 的表达。
维生素 D 可通过干扰 NF-κB 通路减轻 TNF-α诱导的 NP 细胞凋亡。
