Deng Heng, Li Ming, Fang Xiaoli, Tang Kun, Xu Shuqing, Ding Rui, Li Zilong
Department of Anorectal Surgery, Second Affiliated Hospital, Anhui University of Chinese Medicine, Hefei, China.
Department of Anorectal Medicine, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, China.
Front Pharmacol. 2025 Aug 11;16:1646545. doi: 10.3389/fphar.2025.1646545. eCollection 2025.
Although our prior clinical study demonstrated the efficacy of Zuoqing granules (ZQGs) in treating ulcerative colitis (UC), the underlying immunomodulatory mechanisms remained unclear. This study systematically investigated ZQG's therapeutic effects through macrophage polarization modulation and related signaling pathways using both and models.
, dextran sulfate sodium (DSS)-induced UC rats were divided into normal control, DSS model, and ZQG treatment groups (high/medium/low doses). Colon tissues were analyzed using histopathology [hematoxylin and eosin (HE) staining], macrophage phenotyping (CD86/CD206 immunofluorescence and flow cytometry), signaling pathway assessment (PPAR-γ, NF-κB p65, p-NF-κB p65, and STAT1 through Western blot/qPCR), and cytokine profiling (TNF-α, IL-6, IL-10, IL-1β, and IL-4 using ELISA). , lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages were treated with ZQG-conditioned medium and a PPAR-γ antagonist (GW9662) to validate direct effects.
ZQG treatment dose-dependently (1) ameliorated colonic mucosal damage, reducing histological scores by 52% compared to that in the model group, (2) modulated macrophage polarization by increasing the M2 phenotype (CD206 cells, 3.25-fold increase) while decreasing M1 macrophages (CD86 cells, 70% reduction), (3) upregulated PPAR-γ expression (2.0-fold increase) while suppressing NF-κB activation (43% decrease in p-NF-κB) and STAT1 signaling (48% and 40% reductions in protein and mRNA levels, respectively), and (4) rebalanced inflammatory cytokines, with 55%-62% reductions in TNF-α, IL-6, and IL-1β and 185%-210% increases in IL-10 and IL-4. studies further confirmed that ZQG directly shifted macrophage polarization via PPAR-γ, inhibiting M1 polarization (an effect abolished by GW9662).
ZQG ameliorates UC by modulating macrophage plasticity through the PPAR-γ/NF-κB/STAT1 axis, a mechanism validated as PPAR-γ-dependent. These findings elucidate its clinical efficacy and support its use as a multi-target UC therapy.
尽管我们之前的临床研究证明了左清颗粒（ZQGs）治疗溃疡性结肠炎（UC）的疗效，但其潜在的免疫调节机制仍不清楚。本研究使用大鼠和细胞模型，通过巨噬细胞极化调节和相关信号通路系统地研究了ZQG的治疗效果。
将葡聚糖硫酸钠（DSS）诱导的UC大鼠分为正常对照组、DSS模型组和ZQG治疗组（高/中/低剂量）。使用组织病理学[苏木精和伊红（HE）染色]、巨噬细胞表型分析（CD86/CD206免疫荧光和流式细胞术）、信号通路评估（通过蛋白质印迹/qPCR检测PPAR-γ、NF-κB p65、p-NF-κB p65和STAT1）以及细胞因子分析（使用ELISA检测TNF-α、IL-6、IL-10、IL-1β和IL-4）对结肠组织进行分析。此外，用ZQG条件培养基和PPAR-γ拮抗剂（GW9662）处理脂多糖（LPS）刺激的RAW264.7巨噬细胞，以验证直接作用。
ZQG治疗呈剂量依赖性地（1）改善结肠黏膜损伤，与模型组相比，组织学评分降低52%；（2）通过增加M2表型（CD206细胞，增加3.25倍）同时减少M1巨噬细胞（CD86细胞，减少70%）来调节巨噬细胞极化；（3）上调PPAR-γ表达（增加2.0倍），同时抑制NF-κB激活（p-NF-κB减少43%）和STAT1信号传导（蛋白质和mRNA水平分别降低48%和40%）；（4）重新平衡炎性细胞因子，TNF-α、IL-6和IL-1β减少55%-62%，IL-10和IL-4增加185%-210%。细胞研究进一步证实，ZQG通过PPAR-γ直接改变巨噬细胞极化，抑制M1极化（GW9662可消除该作用）。
ZQG通过PPAR-γ/NF-κB/STAT1轴调节巨噬细胞可塑性来改善UC，这一机制经细胞实验验证为PPAR-γ依赖性。这些发现阐明了其临床疗效，并支持将其用作UC的多靶点治疗药物。
