| ObjectiveTo investigate the expression of NLRP3 and IL-1β and the distribution characteristics of microecology in spleen-stomach damp-heat syndrome in Helicobacter pylori-related gastric diseases(HPGD),and to explore the pathogenesis of spleen-stomach damp-heat syndrome in HPGD.MethodsIn this study,a total of 107 cases of HPGD were collected,including 49 cases of chronic superficial gastritis(CSG),45 cases of chronic atrophic gastritis(CAG)and 13 cases of gastric cancer(GC).All cases followed the diagnostic criteria of TCM syndrome,of which 70 cases were diagnosed as spleen-stomach damp-heat syndrome and 37 cases were diagnosed as spleen-qi deficiency syndrome.At the same time,10 normal subjects were recruited in the physical examination department.The diagnosis of the disease was confirmed by HE histopathological examination,the infection and degree of Helicobacter pylori were determined by rapid urease and methylene blue staining,the expression of NLRP3 and IL-1β protein was detected by immunohistochemical method,and the composition and structure of gastric mucosal flora were detected by 16 SrDNA high-throughput gene sequencing.The characteristics of gastric mucosal flora in different syndrome types and disease types of HPGD were studied by species abundance,α diversity,β diversity,LDA Effect size analysis and metastat analysis.Results Part Ⅰ Study on Hp infection in different syndromes and types of Helicobacter pylori-related gastric diseases1.General information.:There was no significant difference in age and disease distribution between HPGD spleen-stomach damp-heat syndrome,spleen-qi deficiency syndrome and normal control group.The prevalence rate of spleen-stomach damp-heat syndrome in male was higher than that in spleen-qi deficiency syndrome and normal control group(P < 0.05).There was no significant difference in sex and age between CSG,CAG,GC and normal control group(P < 0.05).2.Comparison of Hp infection in different syndromes and diseases of HPGD: The infection rate of Hp in spleen-qi deficiency group was higher than that in spleen-stomach damp-heat syndrome,but there was no statistical difference(P > 0.05).The Hp infection rate of different diseases showed a trend of CAG > CSG > GC(P > 0.05),There was no statistical difference in the degree of Hp infection among different syndrome types and disease types.3.Comparison of inflammation in different syndromes and diseases of HPGD: The incidence of inflammation in spleen-qi deficiency syndrome group was higher than that in spleen-stomach damp-heat syndrome group,but there was no statistical difference(P >0.05).The incidence of inflammation of different diseases was GC > CAG > CSG(P <0.05).There was no statistical difference in the degree of inflammation among different syndrome types and diseases.Part Ⅱ The expression of NLRP3 and IL-1β in different syndromes and types of Helicobacter pylori-related gastric diseases1.There were significant differences in the expression of NLRP3 and IL-1β between spleen-stomach damp-heat syndrome,spleen-qi deficiency syndrome and normal control group.After compared with each other,we found that the expression of NLRP3 and IL-1βin spleen-stomach damp-heat syndrome was higher than that in normal control group(P <0.05),but there was no statistical difference between spleen-stomach damp-heat syndrome and spleen qi deficiency syndrome(P > 0.05).2.There were statistical differences in the expression of NLRP3 and IL-1β among normal control group(NC),CSG,CAG and GC of gastric cancer.After compared with each other,we found that the expression of NLRP3 and IL-1β in CAG was higher than that in NC(P <0.05).3.There was no significant correlation between the degree of intestinal metaplasia and the expression of NLRP3 and IL-1β in Helicobacter pylori-related gastric diseases.Part III Study on the changes of gastric microecology in different syndromes and types of Helicobacter pylori-related gastric diseases1.Analysis of gastric mucosal flora of HPGD spleen-stomach damp-heat syndrome.(1)Species abundance analysis: At the Pylum level,Proteobacteria,Firmicutes,Bacteroidetes,Actinobacteria and Fusobacteria are the bacteria enriched together in spleen-stomach damp-heat syndrome,spleen-qi deficiency syndrome and normal control group.(2)Diversity analysis: There was significant difference in shannon and simpson index among spleen-stomach damp-heat syndrome group,spleen-qi deficiency syndrome group and normal control group(P < 0.05).There was no significant difference in sobs index,chao index and ace index among the three groups.Among them,there was significant statistical difference in shannon and simpson index between spleen-stomach damp-heat syndrome and normal control group,but there was no statistical difference in each index between spleen-stomach damp-heat syndrome and spleen qi deficiency syndrome.The results showed that there were significant differences in species abundance and diversity among spleen-stomach damp-heat syndrome group,spleen-qi deficiency syndrome group and normal control group,and the species abundance and diversity of spleen-stomach damp-heat syndrome were lower than those in normal control group.(3)Significant difference of microflora in spleen-stomach damp-heat syndrome:(1)Spleen-stomach damp-heat syndrome and normal control group: At genus level,the dominant flora in the spleen-stomach damp-heat syndrome group was Helicobacter pylori,and the dominant bacteria in the normal control group were Rothia,Neisseria and Enhydrobacter bacteria.(2)Spleen-stomach damp-heat syndrome and spleen-qi deficiency syndrome group: At genus level,the dominant flora of spleen-qi deficiency syndrome group are Clostridium,Lactobacillus and Alloprevotella,while spleen-stomach damp-heat syndrome group had no dominant bacteria group.2.Analysis of gastric mucosal flora among different types of HPGD:(1)Species abundance analysis: At the pylum level,Firmicutes,Bacteroidetes,Actinobacteria and Fusobacteria are the bacteria co-enriched in NC,CSG,CAG and GC.(2)Diversity analysis: There were significant differences in sobs,chao,ace,shannon and simpson indexes among NC,CSG,CAG and GC groups(P < 0.05).The results showed that there were significant differences in species abundance and diversity among the four groups.There were significant differences in sobs,chao,ace,shannon and simpson indexes between CAG and CSG.There were significant differences in shannon and simpson indexes between GC and NC,CAG and NC(P < 0.05).The results showed that the species abundance and diversity of GC were significantly lower than those of NC,and the species abundance and diversity of CAG were significantly lower than those of NC and CSG.There was no significant difference in species diversity between GC and CAG.(3)Significant difference flora analysis: at genus level,the dominant flora of CSG were Neisseria,Alloprevotella,Rothia,Prevotella and so on.The dominant bacteria of CAG were Corynebacterium＿1 and Faecalibacterium GC.The dominant bacteria were Lactobacillus,Lachnoanaerobaculum,Klebsiella,Endomicrobium,Geodermatophilus,Oleiphilus,Jonesia,Alcanivorax and Thermus.Conclusion1.The high expression of NLRP3 and IL-1β in Helicobacter pylori-related gastric diseases may be one of the inherent pathogenic mechanisms of spleen-stomach damp-heat syndrome.2.The decrease of gastric mucosal flora diversity and species abundance of spleen-stomach damp-heat syndrome may be one of the internal pathogenic mechanisms of Helicobacter pylori-related gastric diseases.Helicobacter pylori,Rothia,Neisseria,Paracoccus and Enhydrobacter may be one of the flora markers that distinguish spleen-stomach damp-heat syndrome from healthy people.The flora of Fusobacterium,Lactobacillus and Alloprevotella may be one of the flora markers to distinguish spleen-stomach damp-heat syndrome from spleen-qi deficiency syndrome.3.The significant decrease of species abundance and microflora diversity in gastric microecology may be one of the main factors leading to the development of HPGD. |
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