School of Basic Medical Science, Zhejiang Chinese Medical University, Hangzhou, China.
Department of Rheumatology, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China.
Front Immunol. 2022 Jul 25;13:943827. doi: 10.3389/fimmu.2022.943827. eCollection 2022.
Lupus nephritis (LN), the most severe organ manifestation of systemic lupus erythematosus (SLE), is generally treated with glucocorticoids (GC) in clinical practice, leading to drug resistance and adverse effects in the long term. Fortunately, the combination of GC and traditional Chinese medical prescriptions can attenuate the adverse effects and improve therapeutic efficiency. Willd (HDW) is one of the most commonly used herbal compounds for LN treatment, which exhibits "heat-clearing" and "detoxification" effects. However, the underlying pharmacological mechanism remains unclear. The present study identified the chemical compounds in HDW extract with UPLC-Q-TOF-MS/MS. A total of 49 components were identified in the HDW extract, and the IL-17 signaling pathway was highly enriched by network pharmacological analysis. MRL/lpr model mice, reflecting the spontaneous development of LN, were used to evaluate the protective activity and investigate the underlying mechanism of the combination treatment. The white blood cell content (WBC), including lymphocytes and neutrophils, cytokines (IL-6, MCP-1, TNF-a), and various autoantibodies (ANA, ab-dsDNA, ab-snRNP/sm) in the blood of MRL/lpr mice were significantly improved by the intragastric administration of HDW. Additionally, the expression of STAT3, IL-17, Ly6G, and MPO in the kidney and neutrophil NETosis were ameliorated with HDW treatment. The pathological and morphological analysis suggested that HDW application could reduce urinary protein levels and inflammatory cell infiltration and inhibit glomerular interstitial cell proliferation. Hence, HDW might ameliorate lupus nephritis by inhibiting IL-6 secretion and STAT3-induced IL-17 expression. The active compounds in HDW were predictively selected with computational methods. The docking affinity of asiatic acid, neoandrographolide to IL-6, glycyrrhetinic acid, oleanolic acid, ursolic acid, and wilforlide A to STAT3 are extremely high. In conclusion, the IL-6 and STAT3/IL-17signaling pathways could be critical regulative targets of HDW on LN.
狼疮性肾炎（LN）是系统性红斑狼疮（SLE）最严重的器官表现，临床上一般采用糖皮质激素（GC）治疗，但长期应用会导致耐药和不良反应。所幸的是，GC 与中药方剂联合应用可以减轻不良反应，提高治疗效率。威灵仙（HDW）是治疗 LN 最常用的草药之一，具有“清热”“解毒”作用，但作用机制尚不清楚。本研究采用 UPLC-Q-TOF-MS/MS 对 HDW 提取物中的化学成分进行了鉴定，共鉴定出 HDW 提取物中的 49 种成分，网络药理学分析表明其主要作用于 IL-17 信号通路。MRL/lpr 模型小鼠可自发发展为 LN，用于评估联合治疗的保护活性和探讨其作用机制。HDW 灌胃给药可显著改善 MRL/lpr 小鼠的白细胞含量（WBC），包括淋巴细胞和中性粒细胞，以及血液中的细胞因子（IL-6、MCP-1、TNF-a）和各种自身抗体（ANA、ab-dsDNA、ab-snRNP/sm）。此外，HDW 治疗可改善肾脏中 STAT3、IL-17、Ly6G 和 MPO 的表达以及中性粒细胞 NETosis。病理和形态学分析表明，HDW 可降低尿蛋白水平、减轻炎症细胞浸润和抑制肾小球间质细胞增殖。因此，HDW 可能通过抑制 IL-6 分泌和 STAT3 诱导的 IL-17 表达来改善狼疮性肾炎。采用计算方法预测 HDW 的活性成分，亚油酸、新穿心莲内酯与 IL-6、甘草次酸、齐墩果酸、熊果酸和威灵仙甲素与 STAT3 的对接亲和力极高。综上所述，IL-6 和 STAT3/IL-17 信号通路可能是 HDW 治疗 LN 的关键调控靶点。
