Abstract: BACKGROUND: Immunosuppression leads to impaired body immune function and aggravates the disease. Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body, but its regulatory mechanism has not been elucidated.
OBJECTIVE: To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. 
METHODS: Twenty-four Sprague-Dawley rats were randomly assigned to three groups: control, model, and herb-partitioned moxibustion groups, with eight rats in each group. The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days. No interventions were administered to the control and model groups after modeling. In contrast, the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan, Shenque, Guanyuan, and Zusanli acupoints using a combination of moxa and herbal cakes, once a day, for 10 consecutive days, with samples being collected the day after the end of the intervention. Peripheral blood was collected from all groups of rats to measure their white blood cell count. RNA-seq was performed on the Illumina sequencing platform, and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. 
RESULTS AND CONCLUSION: Compared with the control group, the model group exhibited a significant decrease in white blood cell count (P < 0.001). RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups, with 1 565 upregulated and 1 461 downregulated. There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group, with 280 upregulated and 255 downregulated. The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group. However, after moxibustion with herbal cakes, these genes were upregulated. Protein-protein interaction network analysis identified 10 core targets, including Oasl, Oas2, Isg15, Herc6, Mx2, Helz2, Mx1, Syk, Hspa1a, and Ret. According to GO and KEGG analyses, moxibustion with herbal cakes regulated the body through pathways related to immune response, viruses, angiogenesis, and the autoimmune system. To conclude, there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets, including Oasl, Oas2, Isg15, Herc6, Mx2, Helz2, and Mx1. The intervention exhibits regulatory effects in the pathways related to immune responses, viral activities, and angiogenesis.

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