State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
School of Preclinical Medicine, Chengdu University, Chengdu, 610106, China.
J Ethnopharmacol. 2023 May 10;307:116181. doi: 10.1016/j.jep.2023.116181. Epub 2023 Feb 2.
Huanglian-Houpo Decoction (HLHP), a classical prescription, has been used to treat gastrointestinal diseases for hundreds of years in TCM. However, the effective constituents and underlying mechanisms of HLHP in the treatment of ulcerative colitis (UC) have not been fully investigated.
This study aimed to reveal the potential anti-UC mechanisms of 50% ethanol extraction of HL and HP (EHLHP), combining transcriptomes and network pharmacology, as well as the animal experiment verification.
Primarily, we identified the chemical composition of EHLHP via UPLC-QE-MS analysis. A visualization network with components-targets-pathways on UC treatment were constructed using network pharmacology. And then, the transcriptomics sequencing method was applied to screen out the differentially expressed genes (DEGs) of EHLHP in the treatment of UC. The key targets and pathways of EHLHP were selected by the combination of the network pharmacology and transcriptomics results. Ultimately, the potential mechanisms of EHLHP on DSS-induced UC mice were verified.
A total of 34 components of EHLHP were identified by UPLC-QE-MS analysis. Combined with the analysis of network pharmacology and transcriptomics, there were 262 DEGs between the normal group and the model group, and 151 DEGs between the model group and the EHLHP group. At the same time, there are 79 interaction paths, such as PI3K-Akt signaling pathway, MAPK signaling pathway, etc. These results indicated that the anti-UC mechanisms would be involved in calcium signaling pathway, inflammatory signaling pathway (JAK-STAT, TNF-α, cGMP-PKG) and immune regulation (IL-17, B cell receptor). After 160 mg/kg and 320 mg/kg EHLHP were given to DSS induced UC mice, these typical symptoms could be significantly alleviated, such as the decrease of DAI value and inflammation level. The IHC staining results of ZO-1, Occludin and Claudin-1 suggested that the intestinal barrier of UC mice was enhanced by EHLHP. The expression of macrophages and immune cells in F4/80, CD11c, Gr-1, NK1.1 by FCM determination indicated that EHLHP could suppress UC by immunosuppression and macrophage polarization M1 to M2.
The potential mechanisms of HLHP extract on DSS-induced UC mice were revealed, by the prediction of integrated analysis of transcriptomes and network pharmacology, and subsequently animal test verification. It would provide a viable strategy to elucidate the mechanisms of TCM classical formula.
黄连-厚朴汤（HLHP）是一种经典的方剂，在中医中已有数百年的历史，用于治疗胃肠道疾病。然而，HLHP 治疗溃疡性结肠炎（UC）的有效成分和潜在机制尚未得到充分研究。
本研究旨在通过转录组学和网络药理学结合动物实验验证，揭示 50%乙醇提取 HL 和 HP（EHLHP）的潜在抗 UC 机制。
首先，我们通过 UPLC-QE-MS 分析鉴定 EHLHP 的化学成分。通过网络药理学构建了一个包含 UC 治疗中成分-靶点-通路的可视化网络。然后，应用转录组测序方法筛选出 EHLHP 治疗 UC 的差异表达基因（DEGs）。通过网络药理学和转录组学结果的结合，选择 EHLHP 的关键靶点和通路。最后，验证 EHLHP 对 DSS 诱导的 UC 小鼠的潜在机制。
通过 UPLC-QE-MS 分析鉴定了 EHLHP 的 34 种成分。结合网络药理学和转录组学分析，正常组与模型组之间有 262 个 DEGs，模型组与 EHLHP 组之间有 151 个 DEGs。同时，存在 79 个相互作用途径，如 PI3K-Akt 信号通路、MAPK 信号通路等。这些结果表明，抗 UC 机制可能涉及钙信号通路、炎症信号通路（JAK-STAT、TNF-α、cGMP-PKG）和免疫调节（IL-17、B 细胞受体）。EHLHP 以 160mg/kg 和 320mg/kg 剂量给予 DSS 诱导的 UC 小鼠后，可显著缓解疾病的典型症状，如 DAI 值和炎症水平的降低。免疫组化染色结果显示，ZO-1、Occludin 和 Claudin-1 的表达增加，表明 EHLHP 增强了 UC 小鼠的肠道屏障。FCM 测定的 F4/80、CD11c、Gr-1、NK1.1 中巨噬细胞和免疫细胞的表达表明，EHLHP 可通过抑制 UC 中的免疫抑制和巨噬细胞极化 M1 向 M2 来抑制 UC。
通过转录组学和网络药理学的综合分析预测，并随后进行动物试验验证，揭示了 HLHP 提取物对 DSS 诱导的 UC 小鼠的潜在机制。这为阐明中药经典方剂的机制提供了一种可行的策略。
