School of Public Health, Zhengzhou University, Zhengzhou 450001, China.
Int J Mol Sci. 2023 Sep 5;24(18):13703. doi: 10.3390/ijms241813703.
T-2 toxin could induce bone damage. But there is no specific mechanism about the long non-coding RNAs (lncRNAs) involved in T-2 toxin-induced articular cartilage injury. In this study, 24 SD rats were randomly divided into a control group and a T-2 group, which were administered 4% absolute ethanol and 100 ng/g · bw/day of T-2 toxin, respectively. After treatment for 4 weeks, safranin O/fast green staining identified the pathological changes in the articular cartilage of rats, and immunofluorescence verified the autophagy level increase in the T-2 group. Total RNA was isolated, and high-throughput sequencing was performed. A total of 620 differentially expressed lncRNAs (DE-lncRNAs) were identified, and 326 target genes were predicted. Enrichment analyses showed that the target genes of DE-lncRNAs were enriched in the autophagy-related biological processes and pathways. According to the autophagy database, a total of 23 autophagy-related genes were identified, and five hub genes (Foxo3, Foxo1, Stk11, Hdac4, and Rela) were screened using the Maximal Clique Centrality algorithm. The Human Protein Atlas database indicated that Rela and Hdac4 proteins were highly expressed in the bone marrow tissue, while Foxo3, Foxo1, and Stk11 proteins were reduced. According to Enrichr, etoposide and diatrizoic acid were identified as the key drugs. The real-time quantitative PCR results were consistent with the RNA sequencing (RNA-Seq) results. These results suggested that autophagy was involved in the rat articular cartilage lesions induced by T-2 toxin. The lncRNAs of NONRATG014223.2, NONRATG012484.2, NONRATG021591.2, NONRATG024691.2, and NONRATG002808.2, and their target genes of Foxo3, Foxo1, Stk11, Hdac4, and Rela, respectively, were the key regulator factors of autophagy.
T-2 毒素可诱导骨损伤。但 T-2 毒素诱导关节软骨损伤涉及的长链非编码 RNA（lncRNA）的具体机制尚不清楚。在这项研究中，将 24 只 SD 大鼠随机分为对照组和 T-2 组，分别给予 4%的无水乙醇和 100ng/g·bw/天的 T-2 毒素。治疗 4 周后，番红 O/快绿染色鉴定大鼠关节软骨的病理变化，免疫荧光验证 T-2 组自噬水平升高。提取总 RNA，进行高通量测序。共鉴定出 620 个差异表达的 lncRNA（DE-lncRNA），预测出 326 个靶基因。富集分析表明，DE-lncRNA 的靶基因富集在自噬相关的生物过程和途径中。根据自噬数据库，共鉴定出 23 个自噬相关基因，并用最大团中心度算法筛选出 5 个关键基因（Foxo3、Foxo1、Stk11、Hdac4 和 Rela）。人类蛋白质图谱数据库表明，Rela 和 Hdac4 蛋白在骨髓组织中高表达，而 Foxo3、Foxo1 和 Stk11 蛋白减少。根据 Enrichr，鉴定出依托泊苷和泛影酸为关键药物。实时定量 PCR 结果与 RNA 测序（RNA-Seq）结果一致。这些结果表明，自噬参与了 T-2 毒素诱导的大鼠关节软骨损伤。NONRATG014223.2、NONRATG012484.2、NONRATG021591.2、NONRATG024691.2 和 NONRATG002808.2 的 lncRNA 及其靶基因 Foxo3、Foxo1、Stk11、Hdac4 和 Rela 分别是自噬的关键调节因子。
