Liu Lele, Zhang Hua, Jin Baiming, Li Haonan, Zheng Xiujuan, Li Xuying, Li Mengyuan, Li Mingqi, Nian Shijing, Wang Kewei
Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, Harbin, 150081, China; National Health Commission & Education Bureau of Heilongjiang Province, Key Laboratory of Etiology and Epidemiology, Harbin Medical University (23618504), Harbin, 150081, China; Heilongjiang Provincial Key Laboratory of Trace Elements and Human Health, Harbin Medical University, Harbin, 150081, China; Institute of Cell Biotechnology, China and Russia Medical Research Center, Harbin Medical University, Harbin, 150081, China.
Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, Harbin, 150081, China; Department of Preventive Medicine, Qiqihar Medical University, Qiqihar, 161006, China; National Health Commission & Education Bureau of Heilongjiang Province, Key Laboratory of Etiology and Epidemiology, Harbin Medical University (23618504), Harbin, 150081, China; Heilongjiang Provincial Key Laboratory of Trace Elements and Human Health, Harbin Medical University, Harbin, 150081, China; Institute of Cell Biotechnology, China and Russia Medical Research Center, Harbin Medical University, Harbin, 150081, China.
Toxicon. 2023 Mar 15;225:107049. doi: 10.1016/j.toxicon.2023.107049. Epub 2023 Feb 14.
T-2 toxin is part of the most toxic fungal secondary metabolites contaminating different kinds of grains. Previous studies have demonstrated that T-2 toxin can influence the survival of chondrocytes and extracellular matrix (ECM) composition. MiR-214-3p is essential for the homeostasis of chondrocytes and ECM. However, the molecular machinery underlying T-2 toxin-induced chondrocyte apoptosis and ECM degradation remain to be elucidated. The present study aimed to investigate the mechanism of miR-214-3p's involvement in T-2 toxin-induced chondrocyte apoptosis and ECM degradation. Meanwhile, the role of the NF-κB signaling pathway was scrutinized. C28/I2 chondrocytes were treated with 8 ng/ml of T-2 toxin for 24 h, after the pretreatment of miR-214-3p interfering RNAs for 6 h. Gene and protein levels involved in chondrocyte apoptosis and ECM degradation were assessed through RT-PCR and Western blotting. The apoptosis rate of chondrocyte was measured by flow cytometry. Results and data indicated that miR-214-3p was decreased in a dose-dependent manner at different concentrations of T-2 toxin. The enhancement of miR-214-3p could alleviate chondrocyte apoptosis and ECM degradation due to T-2 toxin exposure. The upregulation of miR-214-3p was associated with the decreased expression of apoptosis-promoting genes such as Bax and Cleaved-caspase3/caspase3 as well as the increased expression of anti-apoptotic genes such as Bcl2 and Survivin. Furthermore, miR-214-3p stimulated the relative protein expression of collagen Ⅱ but inhibited the expression of MMP13. Overexpressing miR-214-3p could suppress the relative protein expression of IKKβ and phospho-p65/p65, thus blocking the activation of the NF-κB signaling pathway. The study suggested that the miR-214-3p attenuates T-2 toxin-induced chondrocyte apoptosis and ECM degradation through a potential NF-κB signaling pathway.
T-2毒素是污染各类谷物的毒性最强的真菌次生代谢产物之一。先前的研究表明，T-2毒素可影响软骨细胞的存活及细胞外基质（ECM）的组成。miR-214-3p对软骨细胞和ECM的稳态至关重要。然而，T-2毒素诱导软骨细胞凋亡和ECM降解的分子机制仍有待阐明。本研究旨在探讨miR-214-3p参与T-2毒素诱导的软骨细胞凋亡和ECM降解的机制。同时，对核因子κB（NF-κB）信号通路的作用进行了研究。在用miR-214-3p干扰RNA预处理6小时后，将C28/I2软骨细胞用8 ng/ml的T-2毒素处理24小时。通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法评估参与软骨细胞凋亡和ECM降解的基因和蛋白质水平。通过流式细胞术测量软骨细胞的凋亡率。结果表明，在不同浓度的T-2毒素作用下，miR-214-3p呈剂量依赖性降低。miR-214-3p的增强可减轻T-2毒素暴露导致的软骨细胞凋亡和ECM降解。miR-214-3p的上调与促凋亡基因如Bax和裂解的半胱天冬酶3/半胱天冬酶3的表达降低以及抗凋亡基因如Bcl2和生存素的表达增加有关。此外，miR-214-3p刺激了Ⅱ型胶原蛋白的相对蛋白表达，但抑制了基质金属蛋白酶13（MMP13）的表达。过表达miR-214-3p可抑制IKKβ和磷酸化p65/p65的相对蛋白表达，从而阻断NF-κB信号通路的激活。该研究表明，miR-214-3p可能通过NF-κB信号通路减轻T-2毒素诱导的软骨细胞凋亡和ECM降解。
