College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, 410208, Hunan Province, China; Department of Hepatology, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, Hunan Province, China.
College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, 410208, Hunan Province, China.
J Ethnopharmacol. 2024 Jun 28;328:118010. doi: 10.1016/j.jep.2024.118010. Epub 2024 Mar 16.
Traditional herbal pair Paeoniae Radix Rubra (roots of Paeonia lactiflora Pall., Chishao in Chinese) and Aconiti Lateralis Radix Praeparata (lateral roots of Aconitum carmichaelii Debeaux, Fuzi in Chinese) are widely used for the treatment of liver diseases, demonstrating clinical efficacy against acute-on-chronic liver failure (ACLF). As the core drug pair representing the "clearing method" and "warming method" in traditional Chinese medicine (TCM), they align with the TCM syndromic characteristics of ACLF, characterized by a mixture of deficiencies and realities. However, the molecular mechanisms underlying the anti-ACLF effects of Chishao - Fuzi herbal pair remain unclear.
To reveal the immunoinflammatory status of patients with hepatitis B virus-related ACLF (HBV-ACLF) based on macrophage polarization and to explore the mechanism of action of Chishao - Fuzi herbal pair in regulating macrophage polarization against ACLF.
Peripheral blood samples were prospectively obtained from patients with HBV-ACLF, patients with chronic hepatitis B (CHB) in the immunoactive phase and healthy individuals. Flow cytometry, qRT-qPCR, and ELISA were used to reveal the activation status of monocyte-macrophages and the expression differences in related cytokines in the peripheral blood of patients with HBV-ACLF. Then, an ACLF rat model and a macrophage inflammation model in vitro were established. Hematoxylin-eosin staining, immunohistochemical staining, transmission electron microscopy, flow cytometry, western blotting, RT-qPCR, and ELISA were used to observe changes in the expression of M1/M2 macrophage markers and related inflammatory factors after Chishao - Fuzi herbal pair intervention, both in vivo and in vitro.
Patients with HBV-ACLF exhibited an imbalance in M1/M2 macrophage polarization, showing a tendency to activate M1 macrophages with high expression of CD86 and iNOS. This imbalance led to an increase in relevant pro-inflammatory factors (IL-1β, IL-6, TNF-α) and a decrease in anti-inflammatory factors (IL-10, TGF-β, VEGF), exacerbating the uncontrolled immune-inflammatory response. Chishao - Fuzi herbal pair intervention improved liver function, coagulation function, and histopathological injury in ACLF rats. It also partially ameliorated endotoxemia and inflammatory injury in ACLF. The mechanism was to restore the immune-inflammatory imbalance and prevent the exacerbation of inflammatory response to liver failure by promoting macrophage polarization toward M2 anti-inflammatory direction, inhibiting M1 macrophage activation, and increasing the levels of anti-inflammatory factors and decreasing pro-inflammatory factors.
Chishao - Fuzi herbal pair can reduce the systemic inflammatory burden of liver failure by modulating macrophage polarization and restoring ACLF immune-inflammatory imbalance. This study provides new perspectives and strategies for studying HBV-ACLF immune reconstitution and inflammatory response control.
白芍（芍药 Paeonia lactiflora Pall. 的根，赤芍在中国被称为 Chishao）和制川乌（乌头属植物川乌 Aconitum carmichaelii Debeaux 的侧根，在中国被称为 Fuzi）的传统草药组合广泛用于治疗肝脏疾病，对急性肝衰竭（ACLF）具有临床疗效。作为代表中医药“清法”和“温法”的核心药物组合，它们与 ACLF 的中医药证候特征一致，表现为虚实夹杂。然而，赤芍-制川乌草药组合抗 ACLF 的作用机制尚不清楚。
基于巨噬细胞极化，揭示乙型肝炎病毒相关 ACLF（HBV-ACLF）患者的免疫炎症状态，并探讨赤芍-制川乌草药组合调节巨噬细胞极化对抗 ACLF 的作用机制。
前瞻性采集 HBV-ACLF 患者、免疫活跃期慢性乙型肝炎（CHB）患者和健康个体的外周血样本。采用流式细胞术、qRT-PCR 和 ELISA 揭示 HBV-ACLF 患者外周血单核细胞-巨噬细胞的激活状态和相关细胞因子的表达差异。然后，建立 ACLF 大鼠模型和体外巨噬细胞炎症模型。采用苏木精-伊红染色、免疫组织化学染色、透射电镜、流式细胞术、Western blot、RT-qPCR 和 ELISA 观察赤芍-制川乌草药组合干预后 ACLF 大鼠体内和体外 M1/M2 巨噬细胞标志物和相关炎症因子表达的变化。
HBV-ACLF 患者存在 M1/M2 巨噬细胞极化失衡，表现为 CD86 和 iNOS 高表达的 M1 巨噬细胞激活倾向。这种失衡导致相关促炎因子（IL-1β、IL-6、TNF-α）增加和抗炎因子（IL-10、TGF-β、VEGF）减少，加剧了失控的免疫炎症反应。赤芍-制川乌草药组合干预改善 ACLF 大鼠的肝功能、凝血功能和组织病理学损伤，部分改善 ACLF 的内毒素血症和炎症损伤。其机制是通过促进巨噬细胞向抗炎的 M2 极化方向，抑制 M1 巨噬细胞激活，增加抗炎因子水平和减少促炎因子，来恢复免疫炎症失衡，防止炎症反应加剧导致肝衰竭。
赤芍-制川乌草药组合通过调节巨噬细胞极化，恢复 ACLF 免疫炎症失衡，减轻肝衰竭的全身炎症负担。本研究为研究乙型肝炎病毒相关 ACLF 的免疫重建和炎症反应控制提供了新的视角和策略。
