Moxibustion at ST36 activates peritoneal macrophages in CTX-induced immunosuppression in mouse model via IFN-γ

Background: To explored whether moxa cone moxibustion can reduce peritoneal inflammation by increasing the content of peritoneal macrophages and B cells via interferon-gamma. Methods: The mice were randomly divided into three groups with six mice in each group: the control group, model group, and moxibustion group, and the model was established in mice using cyclophosphamide. In the moxibustion group, the mice received moxa cone moxibustion at Zusanli (ST36) for 7 days. Analysis of Peritoneal cell were detected by flow cytometry and immunofluorescence, the protein expression level in the peritoneal fluid were measured with mouse cytokine antibody arrays and verified by enzyme linked immuno sorbent assay test, and RNA-Sequencing was used for peritoneal cell RNA analysis. Results: Our results showed that moxa cone moxibustion could reduce the loss of large peritoneal macrophages and B1 cells (P < 0.05). With the cytokine array analysis and enzyme linked immuno sorbent assay test of peritoneal fluid, we found that IFN-gamma was up-regulated in moxibustion group (P < 0.05). There were 169 genes were down-regulated in the model group and up-regulated in the moxibustion group while 19 genes that were up-regulated in the model group and down-regulated in the moxibustion group via RNA-sequencing. Kyoto Encyclopedia of Genes and Genomes pathway analysis of 188 intersect differentially expressed genes were found that the top 3 pathways with the highest enrichment of up-regulated genes included Hematopoietic cell lineage, Inflammatory bowel disease and Malaria. The differentially expressed genes visualization protein-protein interaction network shows the top 10 genes including Ifng, Grb2, CCR7, CTLA4, CXCR5, Foxp3, kit, PRF1, CD5 and klrg1. Conclusion: These findings showed that moxa cone moxibustion can alleviate chemotherapy-induced diarrhea by reducing the loss of large peritoneal macrophages and B1 cells in the peritoneal cavity, possibly through up-regulating inflammatory bowel disease signaling pathway via interferon-gamma to regulate the survival and function of large peritoneal macrophages and B1 cells.