Effect of mild moxibustion on intestinal microbiota and NLRP6 inflammasome signaling in rats with post-inflammatory irritable bowel syndrome

Figure 1 Diagram of moxibustion in rats. Rats were immobilized on a fixing frame. A: A moxa stick of 5 mm in diameter was ignited and placed 2-3 cm above the bilateral Zusanli (ST26) and Tianshu (ST25) for 10 min per day with 7 d of continuous treatment in the mild moxibustion group; B: The sham moxibustion group was given the same treatment as the mild moxibustion group except that moxibustion was treated with an unlit moxa stick.

Figure 2 A rat post-infectious/post-inflammatory irritable bowel syndrome model established by 2,4,6-trinitrobenzenesulfonic acid induction. A: Gross structure; B: Pathology of the normal control group and model control group on days 7, 14, 21, and 28; C: Abdominal withdrawal reflex scores of the normal and model control groups under different pressure stimulations on day 28 after model generation. ^bP < 0.01 vs normal control group. AWR: Abdominal withdrawal reflex.

Figure 3 Effects of mild moxibustion on abdominal withdrawal reflex scores and low-grade inflammation of the colon in each group. A: Abdominal withdrawal reflex score; B: Observed gross structure; C: Histological score; D: Observed histopathology; E: Pathological score. ^bP < 0.01 vs normal control group; ^dP < 0.01 vs model control group. AWR: Abdominal withdrawal reflex.

Figure 4 Effects of mild moxibustion on colonic ultrastructure of rats with post-infectious/post-inflammatory irritable bowel syndrome.

Figure 5 Effects of mild moxibustion on serum C-reactive protein levels and colonic myeloperoxidase activities in post-infectious/post-inflammatory irritable bowel syndrome rats in each group. A: Serum C-reactive protein levels; B: Myeloperoxidase activities. ^bP < 0.01 vs normal control group; ^dP < 0.01 vs model control group; ^fP < 0.01 vs sham moxibustion group. CRP: C-reactive protein; MPO: Myeloperoxidase.

Figure 6 Effects of mild moxibustion on the relative DNA abundances of targeted bacteria in feces of post-infectious/post-inflammatory irritable bowel syndrome rats in each group. A: Relative DNA abundance of Lactobacillus; B: Relative DNA abundance of Bifidobacterium; C: Relative DNA abundance of Faecalibacterium prausnitzii; D: Relative DNA abundance of Escherichia coli. ^bP < 0.01 vs normal control group; ^cP < 0.05 and ^dP < 0.01 vs model control group; ^fP < 0.01 vs sham moxibustion group.

Figure 7 Effects of mild moxibustion on the mRNA and protein expression of NLRP6, ASC, and Caspase-1 in the colon of post-infectious/post-inflammatory irritable bowel syndrome rats in each group. A: The mRNA expression levels of NLRP6, ASC, and Caspase-1; B: The protein expression of NLRP6, ASC, and Caspase-1. ^aP < 0.05 and ^bP < 0.01 vs normal control group; ^dP < 0.01 vs model control group; ^eP < 0.05 and ^fP < 0.01 vs sham moxibustion group.

Figure 8 Effects of mild moxibustion on the protein expression of NLRP6, ASC, and Caspase-1 in the colon of post-infectious/post-inflammatory irritable bowel syndrome rats. NLRP6, ASC, and Caspase-1 are localized in the cytoplasm of mucosal cells in the mucosal layer of rats in each group.

Figure 9 Effects of mild moxibustion on the mRNA and protein expression of IL-1β and IL-18 in the colon of post-infectious/post-inflammatory irritable bowel syndrome rats in each group. A: The mRNA expression levels of IL-1β and IL-18; B: Protein expression levels of IL-1β and IL-18. ^aP < 0.05 and ^bP < 0.01 vs normal control group; ^dP < 0.01 vs model control group; ^fP < 0.01 vs sham moxibustion group.

Figure 10 Effects of mild moxibustion on the mRNA and protein expression of ITLN1 and RELMβ in the colon of post-infectious/post-inflammatory irritable bowel syndrome rats in each group. A: The mRNA expression levels of ITLN1 and RELMβ; B: Protein expression levels of ITLN1 and RELMβ. ^aP < 0.05 and ^bP < 0.01 vs normal control group; ^dP < 0.01 vs model control group; ^eP < 0.05 and ^fP < 0.01 vs sham moxibustion group.