Ma Chunli, Buren Qiqige, Eerde Mutu, Qing Ming, Bao Xiufang, Zhao Linyun, Cao Lili, Zhang Xin, Bai Qigeqi, Chen Meiling, Hong Mei, Bao Yulong, Lian Hua
School of Basic Medicine, Inner Mongolia Medical University, Hohhot, Inner Mongolia, 010110, People's Republic of China.
Medical Innovation Center for Nationalities, Inner Mongolia Medical University, Hohhot, Inner Mongolia, 010110, People's Republic of China.
Drug Des Devel Ther. 2025 Jul 18;19:6145-6181. doi: 10.2147/DDDT.S527657. eCollection 2025.
To elucidate the mechanisms underlying the therapeutic effects of Tonglaga-5 (TLG5) in the treatment of chronic diarrhea.
The chemical compositions of TLG5 in both in vitro and systemic circulation were analyzed using UPLC-Q/TOF MS. Network pharmacology was applied to predict the therapeutic effects of TLG5 on chronic diarrhea. Molecular docking was utilized to assess the binding affinity between key bioactive compounds and their corresponding targets. A total of 24 male Sprague-Dawley (SD) rats were randomly assigned to three groups: control, model, and TLG5 treatment. Corresponding administration included a standard diet, a high-lactose diet of varying concentrations, and TLG5 by gavage during modeling. Transcriptomic analysis was employed to assess gene expression changes in intestinal tissues. Alterations in the intestinal microbiota were evaluated using 16S rRNA sequencing. qPCR and immunohistochemistry technologies were adopted to analyze the expression of genes and proteins associated with intestinal barrier integrity. ELISA was performed to quantify levels of bile acids and short-chain fatty acids (SCFAs).
UPLC-Q/TOF MS identified 118 compounds in vitro and 52 in the systemic circulation. Network pharmacological analysis revealed that the in vitro and in vivo components of TLG5 were associated with five core targets and five key compounds, respectively. RNA sequencing analysis showed that TLG5 treatment inhibited the inflammatory pathway. 16S rRNA sequencing demonstrated that TLG5 administration led to an upregulation of beneficial microbial populations and a concomitant downregulation of pathogenic bacteria. The TLG5 treatment group presented a reduction in necrotic areas and inflammatory cell infiltration, as well as preservation of the mucosal structure, with a marked decline in inflammatory lesions. Moreover, TLG5 treatment resulted in a significant increase in the levels of SCFAs and bile acids in the cecum.
TLG5 exerts therapeutic effects on chronic diarrhea through multiple mechanisms, including modulation of intestinal microbiota diversity, enhancement of intestinal barrier function, and attenuation of inflammatory responses.
阐明通拉嘎 -5（TLG5）治疗慢性腹泻的作用机制。
采用超高效液相色谱 - 四极杆飞行时间质谱（UPLC-Q/TOF MS）分析TLG5的体外化学成分及体内循环成分。运用网络药理学预测TLG5对慢性腹泻的治疗作用。利用分子对接评估关键生物活性化合物与其相应靶点之间的结合亲和力。将24只雄性Sprague-Dawley（SD）大鼠随机分为三组：对照组、模型组和TLG5治疗组。相应的给药方式包括标准饮食、不同浓度的高乳糖饮食，以及在造模期间通过灌胃给予TLG5。采用转录组分析评估肠道组织中的基因表达变化。利用16S rRNA测序评估肠道微生物群的变化。采用qPCR和免疫组织化学技术分析与肠道屏障完整性相关的基因和蛋白质表达。通过酶联免疫吸附测定（ELISA）定量胆汁酸和短链脂肪酸（SCFAs）的水平。
UPLC-Q/TOF MS在体外鉴定出118种化合物，在体内循环中鉴定出52种化合物。网络药理学分析表明，TLG5的体外和体内成分分别与五个核心靶点和五种关键化合物相关。RNA测序分析显示，TLG5治疗可抑制炎症途径。16S rRNA测序表明，给予TLG5可导致有益微生物群上调，同时病原菌下调。TLG5治疗组坏死面积和炎性细胞浸润减少，黏膜结构得以保留，炎性病变明显减轻。此外，TLG5治疗导致盲肠中SCFAs和胆汁酸水平显著升高。
TLG5通过多种机制对慢性腹泻发挥治疗作用，包括调节肠道微生物群多样性、增强肠道屏障功能和减轻炎症反应。
