Department of Rehabilitation Medicine, Langdong Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region 530028, P.R. China.
Department of Health Care, Nanning Maternal and Child Health Hospital, Nanning, Guangxi Zhuang Autonomous Region 530011, P.R. China.
Mol Med Rep. 2020 Dec;22(6):5012-5022. doi: 10.3892/mmr.2020.11632. Epub 2020 Oct 23.
Apoptosis of nucleus pulposus (NP) cells has an important role in the process of intervertebral disc degeneration (IDD), and the search for novel compounds to prevent apoptosis from occurring is urgently required. In the present study, syringic acid (SyrA) was found to exhibit no cytotoxicity on NP cells, and was able to reverse the cytotoxicity, as well as the abnormal expression of Bcl‑2 and caspase‑3, that were induced by lipopolysaccharide (LPS). The transcriptomes of each group were then analyzed using RNA‑Seq. A total of 65 differentially expressed genes (DEGs) were identified in LPS‑stimulated groups (LPS group vs. control group), 819 DEGs were identified in the SyrA‑reversed groups (SyrA plus LPS group vs. LPS group), and a further 25 DEGs were identified in the SyrA plus LPS group compared with the control group. Reverse transcription‑quantitative PCR validation indicated that the alterations in expression of uroplakin 3B‑like 1 (UPK3BL1), voltage‑dependent calcium channel subunit α‑2/δ‑1 (CACNA2D1) and polo‑like kinase 4 (PLK4) were consistent with the corresponding results of RNA‑Seq, and that these genes were involved in both LPS‑stimulation and SyrA‑reversion processes. Kyoto Encyclopedia of Genes and Genomes analyses indicated that the DEGs in SyrA‑reversed groups were involved in, amongst other pathways, 'Autophagy‑other' and 'Apoptosis‑multiple species'. In conclusion, the addition of SyrA to the NP cells co‑incubated with LPS appeared to help prevent the abnormal expression of mRNAs and apoptosis that had been identified in NP cells incubated with LPS alone. The potential mechanism underlying the reversion of SyrA might be attributed to the regulation of CACNA2D1 and PLK4.
核髓细胞凋亡（NP）在椎间盘退变（IDD）过程中起着重要作用，迫切需要寻找新的化合物来防止凋亡的发生。在本研究中，发现丁香酸（SyrA）对 NP 细胞无细胞毒性，并且能够逆转脂多糖（LPS）诱导的细胞毒性以及 Bcl-2 和 caspase-3 的异常表达。然后使用 RNA-Seq 分析每组的转录组。在 LPS 刺激组（LPS 组与对照组相比）中鉴定出 65 个差异表达基因（DEGs），在 SyrA 逆转组（SyrA 加 LPS 组与 LPS 组相比）中鉴定出 819 个 DEGs，在 SyrA 加 LPS 组与对照组相比）中鉴定出 25 个差异表达基因。逆转录定量 PCR 验证表明，尿路上皮蛋白 3B 样 1（UPK3BL1）、电压依赖性钙通道亚基α-2/δ-1（CACNA2D1）和 Polo 样激酶 4（PLK4）的表达变化与 RNA-Seq 的相应结果一致，这些基因均参与 LPS 刺激和 SyrA 逆转过程。京都基因与基因组百科全书分析表明，SyrA 逆转组中的 DEGs 参与了“自噬-其他”和“凋亡-多种物种”等途径。总之，将 SyrA 添加到与 LPS 共同孵育的 NP 细胞中似乎有助于防止单独用 LPS 孵育的 NP 细胞中鉴定出的异常 mRNA 表达和凋亡。SyrA 逆转的潜在机制可能归因于 CACNA2D1 和 PLK4 的调节。
