Li Nan, Yi Yan-Kui, Zhao Jie, Wang Qiang, Yu Jie-Ying, You Yan-Ting, Zhu Yong-Yan, Liu Yan-Yan, Zhao Xiao-Shan, Pan Dong-Mei
Department of Clinical Basis of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Jinan University, Guangzhou, 510632, China.
Department of Laboratory of Chinese Medicine Pharmaceutics, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China.
Curr Med Sci. 2024 Dec;44(6):1259-1269. doi: 10.1007/s11596-024-2925-8. Epub 2024 Dec 14.
The objective of this study was to explore the therapeutic effects of kaempferol (Kae) on rheumatoid arthritis (RA) and to elucidate the underlying mechanisms.
The collagen-induced arthritis (CIA) model was established using collagen II to induce RA. Mice were treated with Kae at a dose of 25 or 50 mg/kg/day via gavage. Pathological changes in the ankle joint were analyzed. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of inflammatory factors. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to assess the expression of genes associated with the balance of regulatory T (Treg)/T helper 17 (Th17) cells. Flow cytometry was utilized to determine the Treg/Th17 ratio. Furthermore, these techniques were employed to evaluate the impact of miR-34a and Foxp3 dysregulation on cellular functions in RA under the influence of Kae. Dual luciferase reporter assay was conducted to analyze the binding of miR-34a to Foxp3.
Treatment with Kae led to a downregulation of receptor-related orphan receptor gamma t (RORγt) and IL-17 expression, and an upregulation of Foxp3, IL-10, and TGF-β expression in CIA mice. Kae intervention inhibited the production of proinflammatory cytokines and increased the production of anti-inflammatory cytokines. Furthermore, Kae treatment suppressed the expression of miR-34a, which was identified as a target of miR-34a. Finally, Kae regulated Treg/ Th17 balance-related genes and cellular inflammation through the miR-34a/Foxp3 axis.
The study demonstrated that Kae effectively ameliorates CIA in mice by modulating the Treg/Th17 balance and related genes via the miR-34a/Foxp3 axis. These findings suggest that Kae may serve as a promising therapeutic agent for the treatment of RA and for restoring immune homeostasis.
本研究旨在探讨山奈酚（Kae）对类风湿关节炎（RA）的治疗作用，并阐明其潜在机制。
采用Ⅱ型胶原诱导建立胶原诱导性关节炎（CIA）模型以诱导RA。小鼠通过灌胃给予剂量为25或50mg/kg/天的Kae。分析踝关节的病理变化。采用酶联免疫吸附测定（ELISA）法检测炎症因子水平。运用逆转录定量聚合酶链反应（RT-qPCR）评估与调节性T（Treg）/辅助性T细胞17（Th17）细胞平衡相关基因的表达。利用流式细胞术测定Treg/Th17比值。此外，运用这些技术评估在Kae影响下miR-34a和Foxp3失调对RA细胞功能的影响。进行双荧光素酶报告基因测定以分析miR-34a与Foxp3的结合情况。
Kae治疗导致CIA小鼠中受体相关孤儿受体γt（RORγt）和IL-17表达下调，Foxp3、IL-10和TGF-β表达上调。Kae干预抑制促炎细胞因子的产生并增加抗炎细胞因子的产生。此外，Kae治疗抑制了miR-34a的表达，而Foxp3被确定为miR-34a的靶标。最后，Kae通过miR-34a/Foxp3轴调节Treg/Th17平衡相关基因和细胞炎症。
该研究表明，Kae通过miR-34a/Foxp3轴调节Treg/Th17平衡及相关基因，有效改善小鼠的CIA。这些发现提示，Kae可能是一种有前景的治疗RA及恢复免疫稳态的治疗药物。
