Abstract

Context

Our previous studies indicated that Yiguanjian decoction (YGJ) has an anti-hepatic-fibrosis effect and could regulate macrophage status.

Objective

To elucidate the mechanism of YGJ in regulating macrophages.

Materials and methods

Liver cirrhosis was induced by CCl₄ for 12 weeks combined with 2-acetylaminofluorene (2-AAF) for the last 4 weeks in male Wistar rats. YGJ (3.56 mg/kg) orally administered in the last 4 weeks, and SORA (1 mg/kg) as control. In vitro, RAW264.7 cells were treated with lipopolysaccharides (LPSs) to induce macrophage polarization to the M1 phenotype, and they were co-cultured with WB-F344 cells and allocated to M group, YGJ group (2 μg/mL) and WIF-1 group (1 μg/mL) with untreated cells as control. The differentiation direction of WB-F344 cell line was observed in the presence or absence of YGJ. Pathology, fibrosis-related cytokines, macrophage polarization-related components, and Wnt signalling pathway components were detected.

Results

In vivo, the expression levels of α-SMA, Col (1), OV6, SOX9, EpCAM and M1 macrophage-related components (STAT1, IRF3, IRF5, IRF8, SOCS3) significantly decreased in the YGJ group compared with those in the 2-AAF/CCl₄ group (p < 0.01 or 0.05). In vitro, the expression levels of M1 macrophage-related components, including STAT1, NF-κB, IRF3, IRF5, and SOCS3, in RAW264.7 cells decreased significantly in the YGJ group compared with those in the M group (p < 0.05 or p < 0.01). The expression levels of Wnt3A, FZD5, LRP-5/-6, and β-catenin significantly increased in the YGJ group compared with those in the M group (p < 0.05 or p < 0.01). In addition, the expression levels of Wnt-4/-5A/-5B, and FZD2 significantly decreased in the YGJ group compared with those in the M group (p < 0.05 or p < 0.01).

Conclusion

This study suggests that the anti-cirrhosis effect of YGJ is associated with its ability to inhibit macrophage M1-polarization, which provides a scientific basis for the clinical application of YGJ.

中文翻译：

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益冠健汤抑制巨噬细胞M1极化并减轻CCl4/2-AAF诱导的肝纤维化

摘要

语境

我们之前的研究表明，益冠健汤（YGJ）具有抗肝纤维化作用，可以调节巨噬细胞状态。

客观的

阐明YGJ调节巨噬细胞的机制。

材料和方法

_{CCl 4}与 2-乙酰氨基芴 (2-AAF) 联合在雄性 Wistar 大鼠中的最后 4 周诱导肝硬化12 周。YGJ (3.56 mg/kg) 在过去 4 周内口服给药，SORA (1 mg/kg) 作为对照。在体外，RAW264.7细胞用脂多糖（LPSs）诱导巨噬细胞极化为M1表型，与WB-F344细胞共培养，分配到M组、YGJ组（2 μg/mL）和WIF -1 组 (1 μg/mL)，未处理的细胞作为对照。在有无YGJ的情况下观察WB-F344细胞系的分化方向。检测病理、纤维化相关细胞因子、巨噬细胞极化相关成分、Wnt信号通路成分。

结果

在体内，YGJ组α-SMA、Col(1)、OV6、SOX9、EpCAM和M1巨噬细胞相关成分(STAT1、IRF3、IRF5、IRF8、SOCS3)的表达水平较2组显着降低。 -AAF/CCl ₄组（p  < 0.01 或 0.05）。在体外，与M组相比，YGJ组RAW264.7细胞中M1巨噬细胞相关成分STAT1、NF-κB、IRF3、IRF5和SOCS3的表达水平显着降低（p  < 0.05或p  < 0.01)。YGJ组Wnt3A、FZD5、LRP-5/-6、β-catenin的表达水平较M组显着升高（p  < 0.05或p < 0.01)。此外，与M组相比，YGJ组Wnt-4/-5A/-5B、FZD2的表达水平显着降低（p  < 0.05或p  < 0.01）。

结论

本研究提示YGJ的抗肝硬化作用与其抑制巨噬细胞M1-极化的能力有关，为YGJ的临床应用提供了科学依据。