Yiqi Jiedu Huayu decoction inhibits precancerous lesions of chronic atrophic gastritis by inhibiting NLRP3 inflammasome-mediated pyroptosis

Figure 1 Yiqi Jiedu Huayu attenuated pathological changes in gastric mucosa of chronic atrophic gastritis rats with precancerous lesion of gastric cancer. The chronic atrophic gastritis mouse model with precancerous lesion was established by oral administration of N-methyl-N’-nitro-N-nitrosoguanidine and Helicobacter pylori, and they were further treated with Yiqi Jiedu Huayu. A and B: Body weight and daily food intake of rats were presented; C: Hematoxylin-eosin staining was employed to examine pathological changes in gastric mucosa. Black arrows indicated disruption of the various components of the gastric wall structure, green arrows denoted atrophied gastric glands, and yellow arrows pointed to infiltrating inflammatory cells; D: Alcian blue-Periodic acid Schiff staining was performed to analyze gastric intestinal metaplasia; E: Ki67 level in gastric mucosa was assessed using IHC. The measurement data were presented as mean ± SD. n = 10. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. YJHD: Yiqi Jiedu Huayu decoction; HE: Hematoxylin-eosin.

Figure 2 Yiqi Jiedu Huayu increased the secretion of gastric mucosal-related factors and reduced inflammatory factors secretion in chronic atrophic gastritis rats with precancerous lesion of gastric cancer. The gastric mucosal-related factors and inflammatory factors were measure before and after Yiqi Jiedu Huayu decoction treatment in the chronic atrophic gastritis rats with precancerous lesion of gastric cancer. A: Serum interleukin-6 level was determined by enzyme-linked immunosorbent assay (ELISA); B: Serum tumor necrosis factor-α level was determined by ELISA; C: Serum C-reactive protein level was determined by ELISA; D: Serum gastrin level was determined by ELISA; E: Serum pepsin level was determined by ELISA; F: Serum somatostatin was determined by ELISA; G: Serum prostaglandin E2 level was determined by ELISA. The measurement data were presented as mean ± SD. n = 10. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. YJHD: Yiqi Jiedu Huayu decoction; IL-6: Interleukin-6; TNF-α: Tumor necrosis factor-α; CRP: C-reactive protein.

Figure 3 Yiqi Jiedu Huayu inhibited NLRP3 inflammasome-mediated pyroptosis in chronic atrophic gastritis rats with precancerous lesion of gastric cancer. We performed an orally taken of N-methyl-N’-nitro-N-nitrosoguanidine and Helicobacter pylori in rats to induce chronic atrophic gastritis with precancerous lesion combined with Yiqi Jiedu Huayu decoction treatment. A and B: Serum interleukin (IL)-18 and IL-1β levels were detected using enzyme-linked immunosorbent assay; C: Immunohistochemistry was performed to assess NLRP3 level in gastric mucosa; D-F: NLRP3, IL-18 and IL-1β mRNA levels in gastric mucosa were measured using quantitative real-time polymerase chain reaction. The measurement data were presented as mean ± SD. n = 10. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. YJHD: Yiqi Jiedu Huayu decoction; IL: Interleukin.

Figure 4 Yiqi Jiedu Huayu decoction inhibited NLRP3 inflammasome activation by inactivating TLR4/NF-κB and IL-6/STAT3 pathways. A: We performed an orally taken of N-methyl-N’-nitro-N-nitrosoguanidine and Helicobacter pylori in rats to induce chronic atrophic gastritis with precancerous lesion combined with Yiqi Jiedu Huayu decoction treatment, and TLR4, p-p65, p65, p-STAT3 and STAT3 protein levels in gastric mucosa were assessed by western blot (n = 10); B-D: The interaction between NLRP3 promoter between H3K9ac/p-STAT3/p-65 was analyzed using chromatin immunoprecipitation assay. The measurement data were presented as mean ± SD. All data were obtained from at least three replicate experiments. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. YJHD: Yiqi Jiedu Huayu decoction; IgG: immunoglobulin G.