Guangdong Food and Drug Vocational College, Guangzhou, China.
South China Normal University Hospital, Guangzhou, Guangdong, China.
Cartilage. 2023 Sep;14(3):375-385. doi: 10.1177/19476035221114858. Epub 2023 Feb 14.
Cinnamaldehyde (CA) is an active ingredient of Wenyang Tongluo capsule. This study was performed to investigate the function of CA on human chondrocytes.
Different doses of CA were used to treat C28/I2 cells, which were stimulated by interleukin-1β (IL-1β), and then the viability and apoptosis of the cells were examined by cell counting kit-8 and flow cytometry. Interleukin-6 (IL-6), interleukin-20 (IL-20), and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay. Quantitative real-time reverse transcriptase polymerase chain reaction was performed to measure miR-1285-5p, miR-140-5p, IL-20, and high-mobility group box 1 (HMGB1) messenger RNA (mRNA) expression. Western blot assay was performed to detect IL-20, HMGB1, IKBα, phospho-IKBα, IKKα/β, and phospho-IKKα/β expression. Moreover, the relationships between miR-1285-5p and IL-20, as well as miR-140-5p and HMGB1, were validated by dual-luciferase reporter assay.
CA promoted the viability and inhibited the apoptosis of C28/I2 cells stimulated by IL-1β and repressed IL-6, IL-20, and TNF-α levels. CA increased miR-1285-5p and miR-140-5p expression levels. MiR-1285-5p and miR-140-5p promoted the viability and inhibited the apoptosis and inflammation of C28/I2 cells. IL-20 was a target gene of miR-1285-5p, and HMGB1 was a target gene of miR-140-5p. Overexpression of IL-20 or HMGB1 could reverse the effect of CA on C28/I2 cells treated with IL-1β. In addition, HMGB1 increased phospho-IKBα and phospho-IKKα/β expression in IL-1β- and CA-treated C28/I2 cells.
CA protects chondrocytes via regulating miR-1285-5p/IL-20 axis and miR-140-5p/HMGB1/nuclear factor kappa B pathway.
肉桂醛（CA）是温阳通络胶囊的一种有效成分。本研究旨在探讨 CA 对人软骨细胞的作用。
用不同剂量的 CA 处理白细胞介素 1β（IL-1β）刺激的 C28/I2 细胞，然后用细胞计数试剂盒-8 和流式细胞术检测细胞活力和凋亡。酶联免疫吸附试验测定白细胞介素 6（IL-6）、白细胞介素 20（IL-20）和肿瘤坏死因子-α（TNF-α）。实时定量逆转录聚合酶链反应检测 miR-1285-5p、miR-140-5p、IL-20 和高迁移率族蛋白 1（HMGB1）信使 RNA（mRNA）表达。Western blot 检测 IL-20、HMGB1、IKBα、磷酸化 IKBα、IKKα/β 和磷酸化 IKKα/β 表达。此外，通过双荧光素酶报告基因实验验证 miR-1285-5p 与 IL-20 以及 miR-140-5p 与 HMGB1 之间的关系。
CA 促进了 IL-1β 刺激的 C28/I2 细胞的活力，抑制了其凋亡，并抑制了 IL-6、IL-20 和 TNF-α水平。CA 增加了 miR-1285-5p 和 miR-140-5p 的表达水平。miR-1285-5p 和 miR-140-5p 促进了 C28/I2 细胞的活力，抑制了其凋亡和炎症。IL-20 是 miR-1285-5p 的靶基因，HMGB1 是 miR-140-5p 的靶基因。过表达 IL-20 或 HMGB1 可逆转 CA 对 IL-1β 处理的 C28/I2 细胞的作用。此外，HMGB1 增加了 CA 处理的 C28/I2 细胞中磷酸化 IKBα 和磷酸化 IKKα/β 的表达。
CA 通过调节 miR-1285-5p/IL-20 轴和 miR-140-5p/HMGB1/核因子 κB 通路来保护软骨细胞。
