Li Zheng, Ting Jin, Xiaojing Wang, Yingqi Wang, Fengbin Liu, Hong M I
the First Clinical College, Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
Department of Gastroenterology, the First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
J Tradit Chin Med. 2025 Oct;45(5):998-1008. doi: 10.19852/j.cnki.jtcm.2025.05.007.
To explore the effect of Chang'an decoction (, CAD) of ameliorating the immune imbalances in ulcerative colitis (UC) by regulating Rab27 in the P53/high mobility group box 1 pathway.
The functions and important signaling pathways of the Rab27- and UC-related genes were analyzed the use of microarray data from the gene expression omnibus database, gene ontology database, Kyoto encyclopedia of genes and genomes database and gene set enrichment analysis. Dextran sulfate sodium salt-induced colitis mouse model was used to verify the bioinformatics results. Colon length, body weight, and disease activity index were measured. Hematoxylin and eosin staining was applied to validate the histopathology. Tight junction proteins were detected by immunohistochemistry. The proportions of T helper 17 cells (Th17) and regulatory T cells (Treg) in mesenteric lymph nodes were measured flow cytometry. Proinflammatory cytokines like interleukin (IL) 17 (IL-17), IL-21 and IL-22 and anti-inflammatory cytokines like transforming growth factor β and IL-10 in the serum and colon of mice were detected by enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction, respectively. The expression levels of high mobility group box 1 (HMGB1), P53 and phospho- P53 (P-P53) in colonic tissues were detected by immunofluorescence and Western blotting.
Bioinformatics analysis revealed that compared with normal tissues, the expression of Rab27 was significantly increased in UC tissues. Receiver operating characteristic curve showed that Rab27 has the potential to be used as a biomarker for the diagnosis of disease activity. Enrichment analysis showed that UC and Rab27 were mainly associated with small molecule transport, nutrient metabolism, transmembrane transport and the downstream pathway of P53. According to animal experiments, the expression of Rab27 was increased in UC tissues, which aggravated the colonic pathological damage, activated the expression of HMGB1, and also leaded to the imbalance of Th17 and Treg cells. After CAD intervention, Rab27 overexpression, weight loss, colon shortening, and pathological damage were substantial reduced, the expression of tight junction proteins, zona occludens 1 and Occludin were increased. The effect of CAD at high-dose was more obvious. In addition, CAD upgraded the number of Treg cells and the production of TGF-β and IL-10, while decreasing the number of Th17 cells and the expression of inflammatory cytokines (IL-17, IL-21, and IL-22). Moreover, colon inflammation was alleviated by CAD, as indicated by the regulation of HMGB1 and P-P53 expression.
The expression of Rab27, HMGB1 and P-P53 could be decreased by CAD, and the balance of Th17 and Treg cells as well as their related cytokines could be regulated by CAD.
探讨肠安汤（CAD）通过调节P53/高迁移率族蛋白B1（HMGB1）通路中的Rab27改善溃疡性结肠炎（UC）免疫失衡的作用。
利用基因表达综合数据库、基因本体数据库、京都基因与基因组百科全书数据库和基因集富集分析的微阵列数据，分析Rab27和UC相关基因的功能及重要信号通路。采用葡聚糖硫酸钠诱导的结肠炎小鼠模型验证生物信息学结果。测量结肠长度、体重和疾病活动指数。应用苏木精-伊红染色验证组织病理学。通过免疫组织化学检测紧密连接蛋白。采用流式细胞术检测肠系膜淋巴结中辅助性T细胞17（Th17）和调节性T细胞（Treg）的比例。分别采用酶联免疫吸附测定法和定量实时聚合酶链反应检测小鼠血清和结肠中促炎细胞因子如白细胞介素（IL）17（IL-17）、IL-21和IL-22以及抗炎细胞因子如转化生长因子β和IL-10。通过免疫荧光和蛋白质印迹法检测结肠组织中HMGB1、P53和磷酸化P53（P-P53）的表达水平。
生物信息学分析显示，与正常组织相比，UC组织中Rab27的表达显著增加。受试者工作特征曲线显示，Rab27有潜力用作疾病活动诊断的生物标志物。富集分析表明，UC与Rab27主要与小分子转运、营养代谢、跨膜转运以及P53的下游通路相关。动物实验表明，UC组织中Rab27表达增加，加重结肠病理损伤，激活HMGB1表达，还导致Th17和Treg细胞失衡。CAD干预后，Rab27过表达、体重减轻、结肠缩短和病理损伤明显减轻，紧密连接蛋白闭合蛋白1和闭合蛋白的表达增加。高剂量CAD的作用更明显。此外，CAD增加Treg细胞数量以及TGF-β和IL-10的产生，同时减少Th17细胞数量和炎性细胞因子（IL-17、IL-21和IL-22）的表达。而且，CAD可减轻结肠炎症，这通过调节HMGB1和P-P53表达得以体现。
CAD可降低Rab27、HMGB1和P-P53的表达，并调节Th17和Treg细胞及其相关细胞因子的平衡。
