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August 17, 2025Open Access

One mechanism of spleen-kidney yang deficiency IBS-D: intestinal microbiota affect ATPase

QLQi LongShanghai Jiao Tong University LLLiwen LiHunan University of Traditional Chinese Medicine NDNa DengLaboratoire de Chémo-biologie synthétique et thérapeutique

Key Points

Reduced Na + -K + -ATPase and Ca 2 + -Mg 2 + -ATPase activity linked to spleen-kidney yang deficiency IBS-D development, indicating energy metabolism's importance.
ELISA tests showed significant decreases in ATPase activity among model mice with IBS-D, correlating with altered intestinal microbiota.
The use of adenine and Folium senna combined with stress methods effectively established the spleen-kidney yang deficiency IBS-D model in mice.
Correlation analysis indicated specific microbial interactions with digestive enzyme activities, highlighting potential therapeutic targets.

Abstract

Background This study aimed to investigate the effects of adenine and Folium senna combined with restraint tail-clamping stress method on the intestinal microbiota of mice with spleen-kidney yang deficiency type diarrhea predominant irritable bowel syndrome (IBS-D) from the perspective of energy metabolism. Methods Twenty SPF-grade female mice were randomly divided into two groups, normal group (CN group) and model group (MD group), with 10 mice per group. A spleen-kidney yang deficiency IBS-D model was established using adenine and Folium senna combined with restraint tail-clamping stress. After model establishment, enzyme-linked immunosorbent assay (ELISA) was used to detect the activity of Na + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase; spectrophotometry was used to measure intestinal digestive enzyme activity; fluorometric diacetate (FDA) hydrolysis spectrophotometry was used to detect intestinal microbial activity; and 16S rRNA sequencing was performed to identify intestinal microbiota in the intestinal contents. Results (1) ELISA showed a significant decrease in Na + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase activity in spleen-kidney yang deficiency IBS-D model mice. (2) Intestinal enzyme activity analysis revealed increased lactase and amylase activity, and decreased sucrase activity in model mice. (3) Intestinal microbial activity analysis demonstrated significantly enhanced in microbial activity in spleen-kidney yang deficiency IBS-D model mice. (4) Correlation analysis revealed a significant positive correlation between Heminiphilus, Clostridium, Desulfovibrio and Ca 2+ -Mg 2+ -ATPase, a significant positive correlation between Heminiphilus and Na + -K + -ATPase, a significant positive correlation between Pediococcus and sucrase, and a significant negative correlation between Clostridium and sucrase. Conclusion Reduced Na + -K + -ATPase and Ca 2 + -Mg 2 + -ATPase activities, increased sucrase and amylase activities, decreased lactase activity, and enhanced intestinal microbial activity may constitute important mechanisms underlying the development of spleen-kidney yang deficiency IBS-D.

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