Abstract：
The immunosuppressive microenvironment is a vital factor for the hepatocellular carcinoma (HCC) progression. However, effective treatment is lacking at current. Shenlian decoction (SLD) is a registered herbal therapy for the HCC treatment, but the underlying mechanism of SLD remains largely elusive. Here, we aimed to explore the anti-tumor effect of SLD in the treatment of HCC. SLD was intragastrically given after the tumor initiation in β-catenin/C-Met or DEN and CCl4 induced HCC mouse model. The tumor growth levels were evaluated by liver weight and histological staining. The tumor-infiltrating immune cells were detected by immunological staining and flow cytometry. The mechanism of the SLD was detected by non-targeted proteomics and verified by a cell co-culture system. The result showed that SLD significantly attenuated HCC progression. SLD promoted macrophage infiltration and increased the M1/M2 macrophage ratio within the tumor tissues. Non-targeted proteomics showed the inhibition of complement C5/C5a signaling is the key mechanism of SLD. Immunological staining showed SLD inhibited C5/C5a expression and C5aR1+ macrophage infiltration. The suggested mechanism was demonstrated by application of C5aR1 inhibitor, PMX-53 in mouse HCC model. Hepatoma cell-macrophage co-culture showed SLD targeted hepatoma cells and inhibited the supernatant-induced macrophage M2 polarization. SLD inhibited AMPK/p38 signaling which is an upstream mechanism of C5 transcription. In conclusion, we found SLD relieved immune-suppressive environment by inhibiting C5 expression. SLD could suppress the C5 secretion in hepatoma cells via inhibition of AMPK/p38 signaling. We suggested that SLD is a potential herbal therapy for the treatment of HCC by alleviating immune-suppressive status.
摘要：
免疫抑制微环境是肝细胞癌（HCC）进展的重要因素。然而,目前缺乏有效的治疗。参连汤（SLD）是一种用于HCC治疗的注册草药疗法，但SLD的潜在机制在很大程度上仍然难以捉摸。这里,我们旨在探讨SLD在HCC治疗中的抗肿瘤作用。在β-catenin/C-Met或DEN和CCl4诱导的HCC小鼠模型中，肿瘤开始后胃内给予SLD。通过肝脏重量和组织学染色评估肿瘤生长水平。通过免疫染色和流式细胞术检测肿瘤浸润免疫细胞。通过非靶向蛋白质组学检测SLD的机制，并通过细胞共培养系统进行验证。结果显示,SLD显著减弱HCC的进展。SLD促进肿瘤组织内巨噬细胞浸润并增加M1/M2巨噬细胞比例。非靶向蛋白质组学研究表明抑制补体C5/C5a信号传导是SLD的关键机制。免疫染色显示SLD抑制C5/C5a表达和C5aR1+巨噬细胞浸润。通过应用C5aR1抑制剂，小鼠HCC模型中的PMX-53。肝癌细胞-巨噬细胞共培养显示SLD靶向肝癌细胞并抑制上清液诱导的巨噬细胞M2极化。SLD抑制作为C5转录的上游机制的AMPK/p38信号传导。总之,我们发现SLD通过抑制C5表达缓解免疫抑制环境。SLD可以通过抑制AMPK/p38信号传导来抑制肝癌细胞的C5分泌。我们建议SLD是一种潜在的草药疗法，可通过减轻免疫抑制状态来治疗HCC。
