Yang Lei, Wang Chaowei, Zhou Jie, Chen Feihong, Liu Lian, Bai Lulu, Wang Xi, Guo Xiong, Yi Shuangqiang
School of Nursing, Health Science Center, Xi'an Jiaotong University, Xi'an, Shaanxi, PR China.
Key Laboratory of Trace Elements and Endemic Diseases of National Health and Family Planning Commission, School of Public Health, Health Science Center, Xi'an Jiaotong University, Xi'an, Shaanxi, PR China.
J Biochem Mol Toxicol. 2025 Apr;39(4):e70231. doi: 10.1002/jbt.70231.
This study aimed to investigate the role of the HDAC4 gene in the pathogenesis of Kashin-Beck disease (KBD) cartilage injury and chondrocyte differentiation induced by T-2 toxin. Immunohistochemistry was used to compare HDAC4 and PTHrP protein expression levels in cartilage from children and adults who have KBD and from respective controls, as well as in cartilage from a rat model exposed to T-2 toxin and selenium deficiency. A KBD cell model was established by exposure to T-2 toxin, and RNA interference was employed to silence HDAC4. Expression levels of mRNA and protein expression levels were subsequently measured before and after HDAC4 gene silencing for genes related to the PTHrP-HDAC4 signaling pathway and cartilage differentiation by real-time quantitative reverse transcription PCR and western blotting. We found that HDAC4 expression levels were not consistent between adult and child chondrocytes. Silencing of HDAC4 resulted in a significant increase in the mRNA expression of Runx2 and PTHrP, and elevated the levels of both the mRNA and protein of MMP13, and increased both the mRNA and protein levels of MEF2C. Notably, following the addition of T-2 toxin, there was a significant increase in Runx2 expression, whereas the levels of MEF2C and MMP13 were markedly decreased in comparison to pre-silencing conditions. These findings indicate that T-2 toxin may influence HDAC4 expression, and the role and regulatory mechanisms of this gene in impairing the differentiation of KBD chondrocytes were explored, thereby offering novel insights into the pathogenesis of KBD.
本研究旨在探讨HDAC4基因在大骨节病（KBD）软骨损伤发病机制以及T-2毒素诱导的软骨细胞分化中的作用。采用免疫组织化学方法比较了患KBD的儿童和成人以及相应对照组的软骨中HDAC4和甲状旁腺激素相关蛋白（PTHrP）的蛋白表达水平，以及暴露于T-2毒素和硒缺乏的大鼠模型软骨中的表达水平。通过暴露于T-2毒素建立KBD细胞模型，并采用RNA干扰使HDAC4沉默。随后通过实时定量逆转录PCR和蛋白质印迹法测量HDAC4基因沉默前后与PTHrP-HDAC4信号通路及软骨分化相关基因的mRNA表达水平和蛋白质表达水平。我们发现，HDAC4在成人和儿童软骨细胞中的表达水平不一致。HDAC4沉默导致Runx2和PTHrP的mRNA表达显著增加，MMP13的mRNA和蛋白质水平升高，MEF2C的mRNA和蛋白质水平也增加。值得注意的是，添加T-2毒素后，Runx2表达显著增加，而与沉默前相比，MEF2C和MMP13的水平明显降低。这些发现表明，T-2毒素可能影响HDAC4表达，并探讨了该基因在损害KBD软骨细胞分化中的作用和调控机制，从而为KBD的发病机制提供了新的见解。
