Liu Xue-Jiao, Ye-Er-Tai Ye-Li-Ya, Jia Yue-Bo, Wu Chen-Heng, Wang Xiang-Xiang, Yang Ke-Ming, Yao Xuan, Ling Jiang-Hong
Department of Gastroenterology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200021, China.
School of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
World J Gastroenterol. 2025 Mar 7;31(9):102256. doi: 10.3748/wjg.v31.i9.102256.
Runchangningshen paste (RCNSP) is a paste made of four medicinal and edible homologous Chinese medicine mixed with honey. It is known for its ability to nourish yin and blood as well as to loosen the bowel to relieve constipation. The pathophysiology of functional constipation (FC) is associated with a reduction in mucin-2 (MUC2) secretion and microbial dysbiosis.
To investigate the underlying mechanism of RCNSP against FC through MUC2 and the gut mucosal microbiota.
Ultra-performance liquid chromatography tandem mass spectrometry characterized RCNSP composition to elucidate the material basis of action. FC model was induced loperamide gavage (16 mg/kg) twice daily for 7 days. Applying defecation function and gastrointestinal motility to assess constipation severity. Hematoxylin and eosin and Alcian blue-periodic acid-schiff staining analyzed colonic mucosal morphology. Transmission electron microscope was used to observe the ultrastructure of goblet cells (GCs). Immunofluorescence colocalization, quantitative PCR, and western blot assessed the impact of RCNSP on gene and protein expression within the NLRP6/autophagy pathway. 16S rDNA was employed to sequence the gut mucosal microbiota.
RCNSP contained 12 components with potential laxative effects. It enhanced defecation function, accelerated gastrointestinal motility, and maintained colonic mucosal integrity. RCNSP treatment significantly increased GC abundance and MUC2 production while preserving GC ultrastructure. At the molecular level, RCNSP enhanced the colocalized expression of key regulatory proteins and modulated mRNA and protein expressions in the NLRP6/autophagy pathway. Through 16S rDNA sequencing analysis, RCNSP significantly altered the mucosal microbiota composition. Specifically, it increased beneficial bacterial strains while reducing harmful ones. Simultaneously, RCNSP reduced butyrate-producing bacteria like , , , and and decreased hydrogen sulfide-producing species, such as . It also reduced bile acid-inhibiting species, such as and while increasing bile acid-producing species, such as .
Our findings suggested that RCNSP ameliorated constipation through a dual mechanism: It stimulated colonic MUC2 secretion by activating NLRP6 inflammasome-mediated autophagy and modulated the composition of the mucosal microbiota.
润肠宁神膏（RCNSP）是一种由四种药食同源的中药与蜂蜜混合制成的膏剂。它以滋阴养血、润肠通便的功效而闻名。功能性便秘（FC）的病理生理学与黏蛋白-2（MUC2）分泌减少和微生物群落失调有关。
通过MUC2和肠道黏膜微生物群研究润肠宁神膏抗功能性便秘的潜在机制。
采用超高效液相色谱串联质谱法对润肠宁神膏的成分进行表征，以阐明其作用的物质基础。通过每日两次灌胃洛哌丁胺（16 mg/kg），持续7天诱导建立功能性便秘模型。应用排便功能和胃肠动力评估便秘严重程度。采用苏木精-伊红染色和阿尔辛蓝-过碘酸-希夫染色分析结肠黏膜形态。透射电子显微镜用于观察杯状细胞（GCs）的超微结构。免疫荧光共定位、定量PCR和蛋白质印迹法评估润肠宁神膏对NLRP6/自噬途径内基因和蛋白质表达的影响。采用16S rDNA对肠道黏膜微生物群进行测序。
润肠宁神膏含有12种具有潜在通便作用的成分。它增强了排便功能，加速了胃肠蠕动，并维持了结肠黏膜的完整性。润肠宁神膏治疗显著增加了杯状细胞的丰度和MUC2的产生，同时保持了杯状细胞的超微结构。在分子水平上，润肠宁神膏增强了关键调节蛋白的共定位表达，并调节了NLRP6/自噬途径中的mRNA和蛋白质表达。通过16S rDNA测序分析，润肠宁神膏显著改变了黏膜微生物群的组成。具体而言，它增加了有益菌菌株，同时减少了有害菌。同时，润肠宁神膏减少了如[具体菌名1]、[具体菌名2]、[具体菌名3]和[具体菌名4]等产丁酸菌，并减少了如[具体菌名5]等产硫化氢菌。它还减少了如[具体菌名6]和[具体菌名7]等抑制胆汁酸的菌，同时增加了如[具体菌名8]等产生胆汁酸的菌。
我们的研究结果表明，润肠宁神膏通过双重机制改善便秘：它通过激活NLRP6炎性小体介导的自噬刺激结肠MUC2分泌，并调节黏膜微生物群的组成。
