Li Min-Yao, Wu Yu-Zhu, Qiu Jian-Guo, Lei Jun-Xuan, Li Mu-Xia, Xu Nan, Liu Yu-Hong, Jin Zhen, Su Zi-Ren, Lee Simon Ming-Yuen, Zheng Xue-Bao, Xiao-Qi Huang
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China; Dongguan Institute of Guangzhou University of Chinese Medicine, Dongguan, China.
Shenzhen Key Laboratory of Hospital Chinese Medicine Preparation, Shenzhen Traditional Chinese Medicine Hospital, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, China.
J Ethnopharmacol. 2023 Jul 15;311:116430. doi: 10.1016/j.jep.2023.116430. Epub 2023 Mar 28.
Huangqin Decoction (HQD), a traditional Chinese medicine (TCM) formula chronicled in Shang Han Lun, is safe and effective for treatment of ulcerative colitis (UC).
To investigate the effect of HQD against dextran sulfate sodium (DSS)-induced UC mice by regulating gut microbiota and metabolites, and further explore the mechanism of fatty acid metabolism on macrophage polarization.
Based on 3% dextran sulfate sodium (DSS)-induced UC mice model, clinical symptoms observation (body weight, DAI, and colon length) and histological inspection were used to evaluate the efficacy of HQD and fecal microbiota transplantation (FMT) from HQD-treated mice. The gut microbiota and metabolites were detected by 16S rRNA sequencing and metabolomics analysis. The parameters of fatty acid metabolism, macrophage polarization, and FFAR1/FFAR4-AMPK-PPARα pathway were analyzed by immunofluorescence analysis, western blotting, and real-time PCR. Then, the effects of FFAR1 and FFAR4 on macrophage polarization were examined by agonists based on LPS-induced RAW264.7 cell model.
The results showed that FMT, like HQD, ameliorated UC by improving weight loss, restoring colon length, and reducing DAI scores and histopathological scores. Besides, HQD and FMT both enhanced the richness of gut microbiota, and modulated intestinal bacteria and metabolites to achieve a new balance. Untargeted metabolomics analysis revealed that fatty acids, especially long-chain fatty acids (LCFAs), dominated in HQD against DSS-induced UC by regulating the gut microenvironment. Further, FMT and HQD recovered the expression of fatty acid metabolism-related enzymes, and simultaneously activated FFAR1/FFAR4-AMPK-PPARα pathway but suppressed NF-κB pathway. Combined with cell experiment, HQD and FMT promoted macrophage polarization from M1 toward M2, which were well associated with anti-inflammatory cytokines and combined with the activated FFAR4.
The mechanism of HQD against UC was related to regulating fatty acid metabolism to mediate M2 macrophage polarization by activating the FFAR4-AMPK-PPARα pathway.
黄芩汤（HQD）是《伤寒论》中记载的一种中药配方，对溃疡性结肠炎（UC）的治疗安全有效。
通过调节肠道微生物群和代谢产物，研究HQD对葡聚糖硫酸钠（DSS）诱导的UC小鼠的影响，并进一步探讨脂肪酸代谢对巨噬细胞极化的作用机制。
基于3%葡聚糖硫酸钠（DSS）诱导的UC小鼠模型，通过观察临床症状（体重、疾病活动指数和结肠长度）和组织学检查来评估HQD和经HQD治疗小鼠的粪便微生物群移植（FMT）的疗效。通过16S rRNA测序和代谢组学分析检测肠道微生物群和代谢产物。通过免疫荧光分析、蛋白质印迹法和实时PCR分析脂肪酸代谢、巨噬细胞极化和游离脂肪酸受体1/游离脂肪酸受体4-AMPK-过氧化物酶体增殖物激活受体α（FFAR1/FFAR4-AMPK-PPARα）通路的参数。然后，基于脂多糖（LPS）诱导的RAW264.7细胞模型，通过激动剂检测FFAR1和FFAR4对巨噬细胞极化的影响。
结果表明，FMT与HQD一样，通过改善体重减轻、恢复结肠长度以及降低疾病活动指数评分和组织病理学评分来改善UC。此外，HQD和FMT均提高了肠道微生物群的丰富度，并调节肠道细菌和代谢产物以实现新的平衡。非靶向代谢组学分析表明，脂肪酸，尤其是长链脂肪酸（LCFAs），通过调节肠道微环境在HQD抗DSS诱导的UC中起主导作用。此外，FMT和HQD恢复了脂肪酸代谢相关酶的表达，同时激活了FFAR1/FFAR4-AMPK-PPARα通路，但抑制了核因子κB（NF-κB）通路。结合细胞实验，HQD和FMT促进巨噬细胞从M1型向M2型极化，这与抗炎细胞因子以及激活的FFAR4密切相关。
HQD抗UC的机制与通过激活FFAR4-AMPK-PPARα通路调节脂肪酸代谢以介导M2巨噬细胞极化有关。
