Abstract：
OBJECTIVE: Kashin-Beck disease (KBD) is an endemic, degenerative, and cartilage-damaging disease for which low selenium and T-2 toxins are considered environmental pathogenic factors. This study aimed to investigate the molecular mechanisms of autophagy in cartilage damage caused by T-2 toxin and the protective effect of chondroitin sulfate A nano-elemental selenium (CSA-SeNP) on the cartilage.
METHODS: KBD chondrocytes and C28/I2 human chondrocyte cell lines were used. T-2 toxin, AKT inhibitor, and CSA-SeNP treatment experiments were conducted separately, with a treatment time of 24 h. Autophagy was monitored using MDC staining, and mRFP-GFP-LC3 adenovirus, respectively. RT-qPCR and western blotting were used to detect the expression of the relevant genes and proteins.
RESULTS: The suppression of autophagy observed in KBD chondrocytes was replicated by applying 10 ng/mL T-2 toxin to C28/I2 chondrocytes for 24 h. The AKT/TSCR/Rheb/mTOR signaling pathway was activated by T-2 toxin, which inhibits autophagy. The supplementation with CSA-SeNP alleviated the inhibition of autophagy by T-2 toxin through the AKT/TSCR/Rheb/mTOR signaling pathway.
CONCLUSIONS: Loss of autophagy regulated by the AKT/TSCR/Rheb/mTOR signaling pathway plays an important role in cartilage damage caused by T-2 toxin. CSA-SeNP supplementation attenuated inhibition of autophagy in chondrocytes by T-2 toxin by modulating this signaling pathway. These findings provide promising new targets for the prevention and treatment of cartilage disease.
METHODS: KBD chondrocytes and C28/I2 human chondrocyte cell lines were used. T-2 toxin, AKT inhibitor, and CSA-SeNP treatment experiments were conducted separately, with a treatment time of 24 h. Autophagy was monitored using MDC staining, and mRFP-GFP-LC3 adenovirus, respectively. RT-qPCR and western blotting were used to detect the expression of the relevant genes and proteins.
RESULTS: The suppression of autophagy observed in KBD chondrocytes was replicated by applying 10 ng/mL T-2 toxin to C28/I2 chondrocytes for 24 h. The AKT/TSCR/Rheb/mTOR signaling pathway was activated by T-2 toxin, which inhibits autophagy. The supplementation with CSA-SeNP alleviated the inhibition of autophagy by T-2 toxin through the AKT/TSCR/Rheb/mTOR signaling pathway.
CONCLUSIONS: Loss of autophagy regulated by the AKT/TSCR/Rheb/mTOR signaling pathway plays an important role in cartilage damage caused by T-2 toxin. CSA-SeNP supplementation attenuated inhibition of autophagy in chondrocytes by T-2 toxin by modulating this signaling pathway. These findings provide promising new targets for the prevention and treatment of cartilage disease.
摘要：
目的：大骨节病（KBD）是一种地方病，退化,和软骨损伤疾病，其中低硒和T-2毒素被认为是环境致病因素。本研究旨在探讨T-2毒素所致软骨损伤中自噬的分子机制及硫酸软骨素A纳米元素硒(CSA-SeNP)对软骨的保护作用。
方法:使用KBD软骨细胞和C28/I2人软骨细胞系。T-2毒素,AKT抑制剂,和CSA-SeNP处理实验分别进行，治疗时间为24小时。使用MDC染色监测自噬，和mRFP-GFP-LC3腺病毒，分别。采用RT-qPCR和蛋白质印迹法检测相关基因和蛋白的表达。
结果：通过将10ng/mL的T-2毒素应用于C28/I2软骨细胞24小时，可以复制在KBD软骨细胞中观察到的自噬抑制。AKT/TSCR/Rheb/mTOR信号通路被T-2毒素激活,抑制自噬。补充CSA-SeNP通过AKT/TSCR/Rheb/mTOR信号通路减轻了T-2毒素对自噬的抑制。
结论：AKT/TSCR/Rheb/mTOR信号通路调节的自噬缺失在T-2毒素引起的软骨损伤中起重要作用。CSA-SeNP补充通过调节该信号通路减弱T-2毒素对软骨细胞自噬的抑制。这些发现为预防和治疗软骨疾病提供了有希望的新靶点。
方法:使用KBD软骨细胞和C28/I2人软骨细胞系。T-2毒素,AKT抑制剂,和CSA-SeNP处理实验分别进行，治疗时间为24小时。使用MDC染色监测自噬，和mRFP-GFP-LC3腺病毒，分别。采用RT-qPCR和蛋白质印迹法检测相关基因和蛋白的表达。
结果：通过将10ng/mL的T-2毒素应用于C28/I2软骨细胞24小时，可以复制在KBD软骨细胞中观察到的自噬抑制。AKT/TSCR/Rheb/mTOR信号通路被T-2毒素激活,抑制自噬。补充CSA-SeNP通过AKT/TSCR/Rheb/mTOR信号通路减轻了T-2毒素对自噬的抑制。
结论：AKT/TSCR/Rheb/mTOR信号通路调节的自噬缺失在T-2毒素引起的软骨损伤中起重要作用。CSA-SeNP补充通过调节该信号通路减弱T-2毒素对软骨细胞自噬的抑制。这些发现为预防和治疗软骨疾病提供了有希望的新靶点。
