School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
Guangxi Scientific Experimental Center of Traditional Chinese Medicine, Guangxi University of Chinese Medicine, Nanning, China.
J Ethnopharmacol. 2019 Mar 1;231:39-49. doi: 10.1016/j.jep.2018.08.033. Epub 2018 Aug 28.
Rhubarb Peony Decoction (RPD) is a formula of traditional Chinese medicine chronicled in Jin Gui Yao Lve, commonly used to treat ulcerative colitis (UC). However, the underlying mechanism of RPD treating UC remains elusive. In our study, we investigated the therapeutic efficacy of RPD and potential mechanism involved in inhibiting dextran sulfate sodium (DSS)-induced ulcerative colitis in mice.
The colitis was induced by DSS in mice for 5 days and estimated body weight loss, disease activity index (DAI) and colon length. Histological changes were observed by H&E staining. The number and abundance of gut mircrobiota were measured with 16 S rDNA sequencing. GC-MS was used to detect the concentration of short chain fatty acids (SCFAs) in cecum. Flow cytometry analyzed the proportion of Th17 and Treg cells in mesenteric lymph nodes (MLNs). IL-17A and Foxp3 in colon were determined by immunohistochemical analyses. The level of cytokine was determined by Multi-Analyte Flow Assay Kit.
Administration of RPD significantly alleviated the pathological changes of UC mice, involving rescued the inflammation-related reduction of colon length, ameliorated body weight loss and damaged tissue. In addition, RPD altered the gut microbiota, involving restored α diversity, increased significantly the abundance of Firmicutes and Actinobacteria, decreased the Proteobacteria and Bacteroidetes. Furthermore, the number of Butyricicoccus pullicaecorum, a butyrate-producing bacterium, were augmented obviously by RPD. Besides, RPD restored the content of SCFA in intestinal tract. Additionally, the proportion of Th17 cells and Treg cells in mesenteric lymph nodes, likewise, the expression of IL-17A and Foxp3 in colon were regulated by RPD, contributing to the restoration of Th17/Treg balance. Moreover, RPD significantly decreased the level of IL-6, TNF-α, IFNγ, IL-10, IL-17A, IL-21, IL-22 in colon, simultaneously increased Treg-related cytokine TGF-β at dose-dependently.
These results demonstrated that RPD had effect on ulcerative colitis, which was related to regulating gut microbiota, especially Butyricicoccus pullicaecorum, and SCFAs to restore the gut Th17/Treg homeostasis.
大黄牡丹汤（RPD）是《金匮要略》中记载的一种中药方剂，常用于治疗溃疡性结肠炎（UC）。然而，RPD 治疗 UC 的潜在机制仍不清楚。在我们的研究中，我们调查了 RPD 的治疗效果以及其在抑制葡聚糖硫酸钠（DSS）诱导的小鼠溃疡性结肠炎中的潜在机制。
通过 DSS 在小鼠中诱导结肠炎 5 天，评估体重减轻、疾病活动指数（DAI）和结肠长度。通过 H&E 染色观察组织学变化。通过 16S rDNA 测序测量肠道微生物的数量和丰度。GC-MS 用于检测盲肠中短链脂肪酸（SCFA）的浓度。通过流式细胞术分析肠系膜淋巴结（MLNs）中 Th17 和 Treg 细胞的比例。通过免疫组织化学分析检测结肠中 IL-17A 和 Foxp3 的水平。通过多分析物流式分析试剂盒测定细胞因子水平。
RPD 给药显著缓解了 UC 小鼠的病理变化，包括挽救了与炎症相关的结肠长度减少、改善了体重减轻和受损组织。此外，RPD 改变了肠道微生物群，包括恢复了 α 多样性，显著增加了厚壁菌门和放线菌门的丰度，减少了变形菌门和拟杆菌门的丰度。此外，RPD 明显增加了丁酸产生菌 Butyricicoccus pullicaecorum 的数量。此外，RPD 还恢复了肠道内 SCFA 的含量。此外，RPD 调节肠系膜淋巴结中 Th17 细胞和 Treg 细胞的比例，以及结肠中 IL-17A 和 Foxp3 的表达，有助于恢复 Th17/Treg 平衡。此外，RPD 显著降低了结肠中 IL-6、TNF-α、IFNγ、IL-10、IL-17A、IL-21 和 IL-22 的水平，同时在剂量依赖性增加 Treg 相关细胞因子 TGF-β。
这些结果表明，RPD 对溃疡性结肠炎有治疗作用，这与调节肠道微生物群有关，特别是与丁酸产生菌 Butyricicoccus pullicaecorum 和 SCFAs 有关，以恢复肠道 Th17/Treg 平衡。
