Zhang Zhiyun, Wang Zhen, Wan Weiping, Li Shumin, Yang Wenzhi, Shi XiNan
Department of Anorectal, the Third Clinical Medical College, Yunnan University of Chinese Medicine, Kunming, 650500, China.
Yunnan University of Chinese Medicine, Kunming, 650500, China.
J Bioenerg Biomembr. 2025 Apr 2. doi: 10.1007/s10863-025-10056-z.
The KunMingShanHaiTang Formula (KMSHTF), adjusted by Professor Zhong Chuanhua for the treatment of ulcerative colitis (UC), is the work of a renowned veteran practitioner of Chinese medicine. However, its specific mechanism remains unknown. Consequently, it is intriguing to investigate the molecular mechanism by which KMSHTF treats UC. To elucidate the mechanism of KMSHTF in the treatment of UC in rats. Initially, the active ingredients and key target genes of KMSHTF in treating UC were analyzed using network pharmacology. Protein-Protein interaction and gene enrichment analyses were performed to predict key targets and pathways. Subsequently, UC rats were treated with KMSHTF, and the expression proteins in intestinal tissue were detected. Finally, the active compounds of KMSHTF intreating ulcerative colitis were further screened using Molecular Docking, and their pharmacological effects were validated through cell experiments. A total of 47 active compounds and 365 key target genes of KMSHTF for UC treatment were identified through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,along with the GeneCards database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment Analysis revealed that KMSHTF exerted its therapeutic effects on UC through regulating multiple pathways. In this study, the HIF-1α pathway was selected as the main molecular pathway of KMSHTF treating UC, and further validation was conducted through in vivo and in vitro experiments.Animal studies revealed that KMSHTF significantly ameliorated UC symptoms in rats, including diarrhea,rectal bleeding and specific pathological alterations in the intestinal wall. Furthermore, KMSHTF reduced pro-inflammatory cytokines IL-6 and TNF-α, up-regulated IL-4 of M2 macrophages and down-regulated iNOS and IL-1β of M1 macrophages. Additionally, it decreased the expression levels of HKII and GLUT1 related HIF-1α pathway. The three active compounds of KMSHTF, Baicalein, Palmatine and Triptonide-were selected based on their strong binding affinity with HIF-1α and HKII through computational molecular docking. Cellular experiments demonstrated that each of these compounds downregulated the protein expression levels of HIF-1α, HKII, GLUT1 and IL-6 in an intestinal wall cell model. Of Note, Baicalein exhibited the most pronounced effect. However, the overexpression of HIF-1α reversed the Baicalein-induced downregulation of HKII, GLUT1 and IL-6 at the protein level in vitro. KMSHTF may modulate macrophage metabolism to promote M2 polarization through the HIF-1α pathway, thereby contributing to its therapeutic efficacy in ulcerative colitis (UC). Baicalein, Palmatine, and Triptonide are the three core active compounds of KMSHTF that primarily contribute to this hypothesis.
昆明山海丹方（KMSHTF）由钟传华教授调整用于治疗溃疡性结肠炎（UC），是一位著名老中医的成果。然而，其具体机制尚不清楚。因此，研究KMSHTF治疗UC的分子机制很有意思。为阐明KMSHTF治疗大鼠UC的机制。首先，利用网络药理学分析KMSHTF治疗UC的活性成分和关键靶基因。进行蛋白质-蛋白质相互作用和基因富集分析以预测关键靶点和途径。随后，用KMSHTF治疗UC大鼠，并检测肠组织中的表达蛋白。最后，利用分子对接进一步筛选KMSHTF治疗溃疡性结肠炎的活性化合物，并通过细胞实验验证其药理作用。通过中药系统药理学数据库和分析平台以及基因卡数据库，共鉴定出KMSHTF治疗UC的47种活性化合物和365个关键靶基因。基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析表明，KMSHTF通过调节多种途径对UC发挥治疗作用。在本研究中，选择HIF-1α途径作为KMSHTF治疗UC的主要分子途径，并通过体内和体外实验进行进一步验证。动物研究表明，KMSHTF显著改善大鼠UC症状，包括腹泻、直肠出血和肠壁特定病理改变。此外，KMSHTF降低促炎细胞因子IL-6和TNF-α，上调M2巨噬细胞的IL-4，下调M1巨噬细胞的iNOS和IL-1β。此外，它降低了与HIF-1α途径相关的HKII和GLUT1的表达水平。基于通过计算分子对接与HIF-1α和HKII的强结合亲和力，选择了KMSHTF的三种活性化合物，黄芩素、巴马汀和雷公藤内酯醇。细胞实验表明，这些化合物中的每一种都在肠壁细胞模型中下调了HIF-1α、HKII、GLUT1和IL-6的蛋白表达水平。值得注意的是，黄芩素表现出最明显的效果。然而，HIF-1α的过表达在体外蛋白水平上逆转了黄芩素诱导的HKII、GLUT1和IL-6的下调。KMSHTF可能通过HIF-1α途径调节巨噬细胞代谢以促进M2极化，从而有助于其在溃疡性结肠炎（UC）中的治疗效果。黄芩素、巴马汀和雷公藤内酯醇是KMSHTF的三种核心活性化合物，主要促成了这一假设。
