Department of Medical Laboratory Science, College of Medical Science and Technology, I-Shou University, Kaohsiung 82445, Taiwan.
School of Medicine, College of Medicine, I-Shou University, Kaohsiung 82445, Taiwan.
Int J Mol Sci. 2024 Sep 21;25(18):10133. doi: 10.3390/ijms251810133.
Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder characterized by widespread inflammation and multi-organ damage. Toll-like receptor 7 (TLR-7) and autophagy have been implicated in SLE pathogenesis. Rice husk silica liquid (RHSL) has shown potential for modulating inflammatory responses, but its effects on SLE have not been thoroughly investigated. This study aims to evaluate the impact of RHSL on immune responses and autophagy in cell culture experiments, focusing on its effects on TLR-7 signaling, cytokine production, and autophagy modulation. RAW264.7 cells and human peripheral blood mononuclear cells (PBMCs) from healthy donors and SLE patients were used. Cells were stimulated with LPS or TLR-7 agonists and treated with RHSL. Cell viability was assessed, and cytokine levels (TNF-α and IL-6) were measured by ELISA. Autophagy-related proteins (LC3II, ATG5-ATG12) were analyzed by Western blotting. The effect of autophagy inhibition was studied using 3-methyladenine (3-MA). A concentration of 100 μg/mL RHSL did not affect cell viability but significantly reduced the TNF-α production in TLR-7 agonist-stimulated RAW264.7 cells (compared to TLR-7 alone, 3.41 ± 0.54 vs. 6.72 ± 0.07 folds) and PBMCs (compared to TLR-7 alone, 0.97 ± 0.19 vs. 1.40 ± 0.33 folds). RHSL enhanced autophagy, as evidenced by increased LC3II (4.35 ± 1.08 folds) and ATG5-ATG12 (7.07 ± 1.30 folds) conjugation in both RAW264.7 cells and SLE patient-derived PBMCs. The reduction in TNF-α production by RHSL was attenuated by 3-MA, indicating that autophagy plays a role in this process. RHSL also inhibited the translocation of phosphorylated NF-κB into the nucleus, suggesting a mechanism for its anti-inflammatory effects. RHSL exhibits potential as an immunomodulatory agent in SLE by enhancing autophagy and modulating TLR-7 signaling pathways. These findings suggest that RHSL could offer therapeutic benefits for managing inflammatory responses in SLE and warrant further investigation into its clinical applications.
系统性红斑狼疮（SLE）是一种慢性自身免疫性疾病，其特征为广泛的炎症和多器官损伤。Toll 样受体 7（TLR-7）和自噬与 SLE 的发病机制有关。稻壳硅酸钠液（RHSL）已显示出调节炎症反应的潜力，但尚未对其在 SLE 中的作用进行彻底研究。本研究旨在通过细胞培养实验评估 RHSL 对免疫反应和自噬的影响，重点研究其对 TLR-7 信号转导、细胞因子产生和自噬调节的影响。使用来自健康供体和 SLE 患者的 RAW264.7 细胞和人外周血单核细胞（PBMC）进行实验。用 LPS 或 TLR-7 激动剂刺激细胞，并使用 RHSL 处理。通过 ELISA 测量细胞活力和细胞因子水平（TNF-α 和 IL-6）。通过 Western 印迹分析自噬相关蛋白（LC3II、ATG5-ATG12）。通过使用 3-甲基腺嘌呤（3-MA）研究自噬抑制的作用。浓度为 100 μg/mL 的 RHSL 不影响细胞活力，但显著降低 TLR-7 激动剂刺激的 RAW264.7 细胞（与 TLR-7 单独相比，3.41 ± 0.54 倍与 6.72 ± 0.07 倍）和 PBMC（与 TLR-7 单独相比，0.97 ± 0.19 倍与 1.40 ± 0.33 倍）中的 TNF-α 产生。RHSL 通过增加 RAW264.7 细胞和 SLE 患者来源的 PBMC 中的 LC3II（4.35 ± 1.08 倍）和 ATG5-ATG12（7.07 ± 1.30 倍）的连接来增强自噬。3-MA 减弱了 RHSL 对 TNF-α 产生的抑制作用，表明自噬在该过程中起作用。RHSL 还抑制磷酸化 NF-κB 向核内易位，表明其具有抗炎作用的机制。RHSL 作为 SLE 中的免疫调节剂具有潜力，通过增强自噬和调节 TLR-7 信号通路来发挥作用。这些发现表明，RHSL 可为 SLE 中炎症反应的管理提供治疗益处，并需要进一步研究其临床应用。
