Effects Of Miyou Decoction On NLRP3/Caspase-1/GSDMD Cell Pyroptosis Pathway In HP-associated Gastritis Mice

Objective: By establishing mouse models of helicobacterpylori associa ted gastritis(HAG),the effect of Mieyou Decoction on the NLRP3/Ca spase-1/GSDMD pathway in HAG mice was explored.Methods: Thirty SPF BALB/c mice were randomly divided into normal group(9 mice)and model group(21 mice).The normal group was given Bruce Broth by gavage;the model group was given the Helicobacter pylori bacterial suspension by gavage.The normal group and the model group were given intragastric administration once every other day,5 times in total.Bacteria were colonized for 2 weeks after modeling.After 2 weeks,3 mice in the normal group and the modeling model were randomly selected to take gastric tissues for rapid urease detection and Warthin-Starry silver staining for model identification.After successful modeling,the 18 mice in the model were randomly divided into a model group(6 mice),a quadruple drug group(6 mice),and a Mieyou decoction group(6 mice).The normal group and the model group were given 0.2ml normal saline by gavage,the quadruple drug group was given 0.2ml quadruple drug suspension,and the Mieyou decoction group was given 0.2ml of Mieyoutang water decoction.The above four groups were given daily gavage.The stomach was administered once for 14 days without interruption.The mice were sacrificed 30 minutes after the last gavage,and their serum and gastric tissue samples were taken for examination.Elisa method was used to detect the levels of IL-1β and TNF-α in serum samples.The rapid urea test detects HP planting and eradication.HE staining was used to observe the inflammation of gastric mucosa.Western blotting and immunohistochemical methods were used to detect the expression of gastric pyrolysis-related proteins NLRP3,caspase-1,and GSDMD.Results: 1.General observation of mice: After receiving medication,the mice in the Silian drug group and Mieyou decoction group have significantly improved their mental status compared with the model group,their appetite improved,their weight increased,their water intake increased,their activity increased,and their stools formed.2.The results of rapid urease and W-S silver staining indicated that the model mice were successfully planted with HP.3.Rapid urease test after treatment to detect HP eradication in HAG mice.The results showed that the eradication rate of the quadruple drug group was 100%,and the eradication of the Mieyou decoction group was 80%.4.HE staining: The gastric mucosal inflammation of mice in the model group was obvious,and the gastric inflammation of HAG mice was significantly reduced after treatment with quadruple drugs or Mieyou decoction.5.Elisa test: the levels of IL-1β and TNF-α in serum samples of the normal group are low.The expression of IL-1β and TNF-α in the model group was significantly higher than that in the normal group(P<0.01).Compared with the model group,the expression of IL-1β and TNF-α in the Mieyou decoction group and the quadruple drug group were significantly decreased(P<0.01).After treatment with the Mieyou decoction group and the quadruple drug group,the expression of IL-1β and TNF-α inflammatory factors in serum samples were reduced,but there was no significant difference between the two(P>0.05).6.Immunohistochemical method for detection of pyrolysis-related proteins: The pyrolysis-related proteins NLRP3,caspase-1,and GSDMD were low expression in the gastric mucosa of mice in the normal group.The expression of NLRP3,caspase-1and GSDMD protein in the gastric mucosa of the model group was significantly higher than that of the normal group,with a significant difference(P<0.01).Compared with the model group,the protein expressions of NLRP3,caspase-1 and GSDMD in the Mieyou decoction group and the quadruple drug group were significantly decreased,with significant differences(P<0.01).The protein expressions of NLRP3,caspase-1,and GSDMD decreased in the Mieyoutang group and the quadruple drug group,but there was no significant difference between the two(P>0.05).7.Western blotting method for pyrolysis-related proteins:The pyrolysis-related proteins NLRP3,caspase-1,and GSDMD were low in the gastric mucosa of the normal group.The expression of NLRP3,caspase-1 and GSDMD protein in the model group was significantly higher than that in the normal group,and there was a significant difference(P<0.01).Compared with the model group in the Mieyou decoction group and the quadruple drug group,the protein expression of NLRP3,caspase-1and GSDMD decreased significantly(P<0.01).The protein expressions of NLRP3,caspase-1,and GSDMD were decreased in the Mieyou decoction group and the quadruple drug group,but there was no significant difference between the two groups(P>0.05).Conclusion: HP infection of GES-1 cells can induce pyrolysis and ca use severe inflammation.After treatment with Mieyou decoction,the g eneral condition of model mice was improved,the expression of NLR P3,caspase-1,and GSDMD protein in gastric tissue decreased,and ga stric inflammation was reduced.Therefore,it is speculated that Mieyo u decoction can regulate the pyrolysis pathway of NLRP3/caspase-1/G SDMD cells,inhibit the occurrence of pyrolysis,reduce gastric inflam mation,and provide new targets and new ideas for theprevention and treatment of HAG.