School of Public Health, Health Science Center, Xi'an Jiaotong University, Xian, China.
Key Laboratory of Trace Elements and Endemic Diseases, National Health Commission of the People's Republic of China, Xi'an, China.
Cartilage. 2022 Jul-Sep;13(3):19476035221109226. doi: 10.1177/19476035221109226.
The objective of this study was to determine the matrix metalloproteinase-10 (MMP-10) expression pattern and to assess how it contributes to endochondral osteogenesis in Kashin-Beck disease (KBD).
The cartilages of KBD patients, Sprague-Dawley rats fed with selenium (Se)-deficient diet and/or T-2 toxin, and ATDC5 cells were used in this study. ATDC5 cells were induced into hypertrophic chondrocytes using a 1% insulin-transferrin-selenium (ITS) culture medium for 21 days. The expressions of MMP-10 in the cartilages were visualized by immunohistochemistry. The messenger RNA (mRNA) and protein expression levels were determined by real-time polymerase chain reaction (RT-PCR) and Western blotting. MMP-10 short hairpin RNA (shRNA) was transfected into hypertrophic chondrocytes to knock down the gene expression of MMP-10. Meanwhile, the cell death of MMP-10-knockdown chondrocyte was detected using flow cytometry.
The expression of MMP-10 was decreased in the growth plates of children with KBD. A decreased expression of MMP-10 also was observed in the growth plates of rats fed with an Se-deficient diet and/or T-2 toxin exposure. The mRNA and protein expression levels of MMP-10 increased during the chondrogenic differentiation of ATDC5 cells. MMP-10 knockdown in hypertrophic chondrocytes significantly decreased the gene and protein expression of collagen type II (Col II), Col X, Runx2, and MMP-13. Besides, the percentage of cell apoptosis was significantly increased after MMP-10 knockdown in hypertrophic chondrocytes.
MMP-10 deficiency disrupts chondrocyte terminal differentiation and induces the chondrocyte's death, which impairs endochondral osteogenesis in the pathogenesis of KBD.
本研究旨在确定基质金属蛋白酶-10（MMP-10）的表达模式，并评估其在大骨节病（KBD）中的软骨内成骨作用。
本研究使用了 KBD 患者的软骨、饲以缺硒（Se）饮食和/或 T-2 毒素的 Sprague-Dawley 大鼠以及 ATDC5 细胞。将 ATDC5 细胞用 1%胰岛素-转铁蛋白-硒（ITS）培养基诱导 21 天，使之分化为肥大软骨细胞。采用免疫组织化学法观察软骨中 MMP-10 的表达。采用实时聚合酶链反应（RT-PCR）和 Western blot 法检测 MMP-10 的信使 RNA（mRNA）和蛋白表达水平。将 MMP-10 短发夹 RNA（shRNA）转染至肥大软骨细胞中，以敲低 MMP-10 的基因表达。同时，采用流式细胞术检测 MMP-10 敲低的软骨细胞的死亡情况。
KBD 患儿的生长板中 MMP-10 的表达降低。在饲以缺硒饮食和/或暴露于 T-2 毒素的大鼠的生长板中，MMP-10 的表达也降低。在 ATDC5 细胞的软骨分化过程中，MMP-10 的 mRNA 和蛋白表达水平增加。在肥大软骨细胞中敲低 MMP-10 后，胶原 II（Col II）、Col X、Runx2 和 MMP-13 的基因和蛋白表达均显著降低。此外，在肥大软骨细胞中敲低 MMP-10 后，细胞凋亡的百分比显著增加。
MMP-10 缺乏会破坏软骨细胞的终末分化并诱导其死亡，从而损害 KBD 发病机制中的软骨内成骨作用。
