Department of General Surgery (Department of Coloproctology), The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China; Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China; Biomedical Innovation Center, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Department of Anaesthesiology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Biomol Biomed. 2024 Jan 29;24(4):871-887. doi: 10.17305/bb.2024.10187.
The cellular characteristics of intestinal cells involved in the therapeutic effects of astragaloside IV (AS-IV) for treating slow transit constipation (STC) remain unclear. This study aimed to determine the dynamics of colon tissue cells in the STC model and investigate the effects of AS-IV treatment by single-cell RNA sequencing (scRNA-seq). STC mouse models were developed using loperamide, with subsequent treatment using AS-IV. Colon tissues and feces were collected for scRNA-seq and targeted short-chain fatty acid quantification. We integrated scRNA-seq data with network pharmacology to analyze the effect of AS-IV on constipation. AS-IV showed improvement in defecation for STC mice induced by loperamide. Notably, in STC mice, epithelial cells, T cells, B cells, and fibroblasts demonstrated alterations in cell proportions and aberrant functions, which AS-IV partially rectified. AS-IV has the potential to modulate the metabolic pathway of epithelial cells through its interaction with peroxisome proliferator-activated receptor gamma (PPARγ). AS-IV reinstated fecal butyrate levels and improved energy metabolism in epithelial cells. The proportion of naïve CD4+T cells is elevated in STC, and the differentiation of these cells into regulatory T cells (Treg) is regulated by B cells and fibroblasts through the interaction of ligand-receptor pairs. AS-IV treatment can partially alleviate this trend. The status of fibroblasts in STC undergoes alterations, and the FB_C4_Adamdec1 subset, associated with angiogenesis and the Wingless-related integration (Wnt) pathway, emerges. Our comprehensive analysis identifies perturbations of epithelial cells and tissue microenvironment cells in STC and elucidates mechanisms underlying the therapeutic efficacy of AS-IV.
肠细胞的细胞特征涉及黄芪甲苷（AS-IV）治疗慢传输型便秘（STC）的治疗效果，但目前尚不清楚。本研究旨在通过单细胞 RNA 测序（scRNA-seq）确定 STC 模型中结肠组织细胞的动力学，并研究 AS-IV 治疗的效果。使用洛哌丁胺建立 STC 小鼠模型，随后用 AS-IV 进行治疗。收集结肠组织和粪便进行 scRNA-seq 和靶向短链脂肪酸定量。我们将 scRNA-seq 数据与网络药理学相结合，分析 AS-IV 对便秘的影响。AS-IV 改善了洛哌丁胺诱导的 STC 小鼠的排便。值得注意的是，在 STC 小鼠中，上皮细胞、T 细胞、B 细胞和成纤维细胞的细胞比例和异常功能发生改变，AS-IV 部分纠正了这些改变。AS-IV 有可能通过与过氧化物酶体增殖物激活受体γ（PPARγ）相互作用来调节上皮细胞的代谢途径。AS-IV 恢复了粪便丁酸水平，并改善了上皮细胞的能量代谢。在 STC 中，幼稚 CD4+T 细胞的比例升高，这些细胞向调节性 T 细胞（Treg）的分化受 B 细胞和成纤维细胞通过配体-受体对相互作用的调节。AS-IV 治疗可以部分缓解这种趋势。STC 中成纤维细胞的状态发生改变，出现与血管生成和 Wnt 途径相关的 FB_C4_Adamdec1 亚群。我们的综合分析确定了 STC 中上皮细胞和组织微环境细胞的扰动，并阐明了 AS-IV 治疗疗效的机制。
