Joint Divison of Orthopedic Department, Changzheng Hospital, Shanghai, 200003, China.
Department of Orthopaedics, Fuzhou Second Hospital Affiliated to Xiamen University, No. 47 Shangteng Road, Cangshan District, Fu Zhou, 350007, Fujian, China.
J Orthop Surg Res. 2021 Jan 11;16(1):40. doi: 10.1186/s13018-020-02121-7.
Knee osteoarthritis (KOA) seriously affects the quality of life of KOA patients. This study aimed to investigate whether miR-107 could regulate KOA through pyroptosis to affect collagen protein secreted by chondrocytes through IL-1β.
The proliferation of chondrocytes was detected by CCK-8 assay. RT-qPCR analysis was used to identify miR-107 expression and transfection effects. The expression of Col II, IL-1β, IL-18, and MMP13 in supernatant of chondrocytes or chondrocytes was detected by ELISA assay and western blot analysis. The pyroptosis of chondrocytes was analyzed by TUNEL assay and the expression of pyroptosis-related proteins was analyzed by western blot. Luciferase reporter assay confirmed the relation of miR-107 to caspase-1.
The proliferation of chondrocytes was decreased after LPS induction and further decreased by treatment of ATP. Single LPS treatment for chondrocytes downregulated the Col II expression while upregulated the expression of IL-1β, IL-18, and MMP-13, which was further changed by ATP treatment. miR-107 expression was decreased in chondrocytes induced by LPS and further decreased in chondrocytes induced by LPS and ATP. In addition, miR-107 overexpression increased the proliferation and decreased the pyroptosis of chondrocytes induced by LPS and ATP. miR-107 overexpression upregulated the Col II expression while down-regulated the expression of IL-1β, IL-18, and MMP-13 in supernatant of chondrocytes or chondrocytes induced by LPS and ATP. miR-107 overexpression down-regulated the expression of caspase-1, c-caspase-1, GSDMD-N, and TLR4 in chondrocytes induced by LPS and ATP. Furthermore, miR-107 directly targeted caspase-1.
miR-107 can protect against KOA by downregulating caspase-1 to decrease pyroptosis, thereby promoting collagen protein secreted by chondrocytes by down-regulating IL-1β.
膝骨关节炎(KOA)严重影响 KOA 患者的生活质量。本研究旨在探讨 miR-107 是否可以通过调控 KOA 细胞焦亡,进而通过 IL-1β 影响软骨细胞分泌胶原蛋白。
通过 CCK-8 法检测软骨细胞的增殖情况。采用 RT-qPCR 分析鉴定 miR-107 的表达及转染效果。采用 ELISA 法和 Western blot 分析检测软骨细胞上清液或软骨细胞中 Col II、IL-1β、IL-18 和 MMP13 的表达。采用 TUNEL 法分析软骨细胞焦亡情况,采用 Western blot 法分析焦亡相关蛋白的表达。采用荧光素酶报告实验验证 miR-107 与 caspase-1 的关系。
LPS 诱导后软骨细胞增殖减少,ATP 处理后进一步减少。单独 LPS 处理软骨细胞下调 Col II 表达,上调 IL-1β、IL-18 和 MMP-13 表达,ATP 处理进一步改变这一结果。LPS 诱导的软骨细胞中 miR-107 表达降低,LPS 和 ATP 诱导的软骨细胞中进一步降低。此外,miR-107 过表达增加 LPS 和 ATP 诱导的软骨细胞增殖,减少软骨细胞焦亡。miR-107 过表达上调 LPS 和 ATP 诱导的软骨细胞上清液或软骨细胞中 Col II 的表达,下调 IL-1β、IL-18 和 MMP-13 的表达。miR-107 过表达下调 LPS 和 ATP 诱导的软骨细胞中 caspase-1、c-caspase-1、GSDMD-N 和 TLR4 的表达。此外,miR-107 直接靶向 caspase-1。
miR-107 可以通过下调 caspase-1 减少细胞焦亡,从而促进软骨细胞分泌胶原蛋白,从而对 KOA 起保护作用,进而下调 IL-1β。