State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, Sichuan, China; School of Medical and Life Sciences/Reproductive & Women-Children Hospital, Chengdu University of Traditional Chinese Medicine, China.
School of Medical and Life Sciences/Reproductive & Women-Children Hospital, Chengdu University of Traditional Chinese Medicine, China.
J Ethnopharmacol. 2022 Sep 15;295:115356. doi: 10.1016/j.jep.2022.115356. Epub 2022 May 11.
Huangqin decoction (HQD), composed of Scutellaria(Huangqin), Peony(Shaoyao), Liquorice(Gancao) and Jujube(Dazao), is a traditional Chinese medicine prescription, originated from treatise on Febrile Diseases, has the functions of clearing heat, stopping benefits and relieving pain. It is the original prescription for treating heat and relieving dysentery, and is commonly used in clinic for diarrhea and other diseases. In ulcerative colitis, damp-heat syndrome is the most common. However, its mechanism of action is not completely clear.
The purpose of the research is to investigate the protective effect of HQD on ulcerative colitis rats and the regulation effect of mitochondrial DNA, TLR4, p-Akt, p-PI3K protein and microbiota.
The effects of HQD anti-UC were investigated by fluorescence quantitative PCR, cytokine level and histopathological analysis in DSS-induced ulcerative colitis (UC) rats. The content of mtDNA in colon epithelial cells of rats in each group was detected by fluorescence quantitative PCR, p-PI3K, p-Akt and TLR4 protein expressions in colon tissues of rats in each group were detected by Western blotting. IL-6, IL-17 and IL-23 inflammatory factors were detected by ELISA. The effect of HQD on intestinal microbiota of rats with ulcerative colitis was studied by high-throughput sequencing technology, and the correlation between mtDNA level and inflammatory factors as well as protein expression in colonic epithelium of rats with ulcerative colitis was analyzed by SPSS23.0.
HQD significantly alleviated UC symptoms by improving the mucosal intestinal epithelial cell structure, mental state, hair gloss, fecal occult blood, lamina propria intestinal glands and inflammatory cell infiltration. And HQD reduced the pro-inflammatory cytokines in the colonic epithelium of UC rats Production of IL-6, IL-17 and IL-23. The HE stained section of colon tissue showed a complete intestinal epithelial mucosal layer structure. The structure of epithelial cells was more normal and abundant. There were more goblet cells in lamina propria adenoma, which improved the infiltration of inflammatory cells. HQD significantly inhibited the mtDNA content in rat colonic epithelial tissue, and significantly inhibited the expression of TLR4, p-PI3K and p-Akt inflammatory signaling pathways. The results of the microbiota experiment showed that the abundance of HQD in the phylum Firmicutes increased, and the number of Bacteroides phylum decreased (p < 0.05). At the genus level, HQD significantly increased Lactobacillus and Firmicutes Bacteroides, while Treponema and Bacteroides were significantly reduced (p < 0.05).
HQD has a certain protective effect on rats with damp heat ulcerative colitis. Its mechanism may be related to regulating the expression of p-PI3K, p-Akt and TLR4 proteins, mitochondrial DNA as well as microbiota.
黄芩汤（HQD）由黄芩、芍药、甘草和大枣组成，是一种传统的中药方剂，源自《伤寒论》，具有清热、止利、止痛的功效。它是治疗热痢的原方，临床上常用于治疗腹泻等疾病。在溃疡性结肠炎中，湿热证最为常见。然而，其作用机制尚不完全清楚。
本研究旨在探讨 HQD 对溃疡性结肠炎大鼠的保护作用及其对线粒体 DNA、TLR4、p-Akt、p-PI3K 蛋白和微生物群的调节作用。
通过荧光定量 PCR、细胞因子水平和组织病理学分析，研究 HQD 对 DSS 诱导的溃疡性结肠炎（UC）大鼠的抗 UC 作用。采用荧光定量 PCR 检测各组大鼠结肠上皮细胞中线粒体 DNA 的含量，采用 Western blot 检测各组大鼠结肠组织中 p-PI3K、p-Akt 和 TLR4 蛋白的表达。采用 ELISA 法检测 IL-6、IL-17 和 IL-23 等炎症因子。采用高通量测序技术研究 HQD 对溃疡性结肠炎大鼠肠道微生物群的影响，并采用 SPSS23.0 软件分析 mtDNA 水平与溃疡性结肠炎大鼠结肠上皮炎症因子和蛋白表达的相关性。
HQD 通过改善肠黏膜上皮细胞结构、精神状态、毛发光泽、粪便隐血、固有层肠腺和炎症细胞浸润，显著缓解 UC 症状。HQD 还降低了 UC 大鼠结肠上皮促炎细胞因子 IL-6、IL-17 和 IL-23 的产生。结肠组织 HE 染色切片显示完整的肠上皮黏膜层结构，上皮细胞结构更为正常，固有层腺瘤中杯状细胞增多，炎症细胞浸润减轻。HQD 显著抑制大鼠结肠上皮组织中线粒体 DNA 的含量，并显著抑制 TLR4、p-PI3K 和 p-Akt 炎症信号通路的表达。微生物实验结果表明，HQD 可增加厚壁菌门的丰度，减少拟杆菌门的数量（p<0.05）。在属水平上，HQD 显著增加了乳杆菌属和厚壁菌门双歧杆菌属，而减少了栖瘤胃菌属和拟杆菌属（p<0.05）。
HQD 对湿热型溃疡性结肠炎大鼠具有一定的保护作用，其机制可能与调节 p-PI3K、p-Akt 和 TLR4 蛋白、线粒体 DNA 以及微生物群有关。
