Atractyloside-A ameliorates spleen deficiency diarrhea in mice by modulating Lactobacillus johnsonii to enhance butyrate production, activating GPR43, and inhibiting the assembly of NLRP3

Background Spleen-deficiency diarrhea (SDD) is recognized in Traditional Chinese Medicine (TCM) as a prevalent gastrointestinal ailment, primarily characterized by chronic and persistent diarrhea, akin to classifications in Western medicine. The disruption of the intestinal barrier and the appearance of intestinal inflammation are the direct causes of the occurrence of SDD. Therefore, restoring the integrity of the intestinal barrier and reducing inflammation is an important strategy for relieving SDD. Atractyloside-A (AA), a major bioactive compound derived from the post-processing of Atractylodes lancea (Thunb.) DC., is recognized as a crucial component that enhances the spleen-strengthening effects following the processing of this herb. Previous studies have indicated that AA is effective in treating SDD through the modulation of gut microbiota; however, the specific molecular mechanisms underlying this effect remain unclear. This study aimed to elucidate the molecular pathways by which AA exerts its therapeutic effects on SDD. Results This study showed that AA significantly increased the abundance of Lactobacillus johnsonii , promoted butyrate production, enhanced GPR43 expression, and modulated the TLR4/NF-κB signaling pathway. This modulation inhibited the assembly of the NLRP3 inflammasome, alleviated inflammation, reversed mucin synthesis damage, and reduced the pathological symptoms associated with SDD. Moreover, administration of Lactobacillus johnsonii , NaB, and SCFAs individually resulted in significant improvements in various SDD-related phenotypes. However, the beneficial effects of AA and NaB were abolished following the administration of AAV-shGPR43. Conclusion AA enhances butyrate production by regulating Lactobacillus johnsonii , activates GPR43, and inhibits the assembly of the NLRP3 inflammasome through modulation of the TLR4/NF-κB pathway, thereby ameliorating SDD in mice.