Study Of Expression Of NLRP3Inflammasomes In The Mice Of Helicobacter Pylori Infection

Objectives:To construct Helicobacter pylori infected C57BL/6mouse model, monitor theexpression levels of NLRP3inflammsomes signal pathway related moleculars in thegastric tissues, systematically analyze the role of NLRP3inflammsomes signalpathway and the moleculars in inflammatory response caused by Helicobacter pyloriinfection.Methods:6-8weeks old C57BL/6mice were randomly divided into3groups as control,infection and treatment group. The mice of infection group were challenged with H.pylori, while control group with PBS for five times every48hours. Four weeks afterchallenge, some mice were sacrificed to isolate gastric tissue and culture for H. pylorito identify the infection was successful. At the same time, some gastritis mice weretreated with pantoprazole, darithromycin, as well as ampicillin. The isolated gastrictissue and blood of mice were used for the following studies:(1) Histopathology observe the inflammatory response in gastric tissue byhematoxylin and eosin stain;(2) Monitor mRNA levels of NLRP3, ASC, caspase-1, IL-1β, IL-18, IL-33in micegastric tissue by RT-PCR;(3) Detect cytokine levels of IL-1β, IL-18and IL-33in mice serum by ELISA;(4) Analyze expression of caspase-1in gastric tissue by Western blot.Results:(1) H. pylori infect mice developed severe inflammatory response in gastric tissuecompared with control mice and the extent of inflammation was in a time-dependent maner. Treatment can successfully alleviate the inflammatory response.(1)Gastric tissue of control group is normal, more time did H. pylori infect, theinflammation of gastric tissue of mice become worse, after therapy, the inflammationwill alleviate;(2)Compared with control group and treatment group, the mRNA expression ofNLRP3, ASC, Caspase-1, IL-1β, IL-18, IL-33increased in the gastrictissue;(P<0.05)(3)Contents of IL-1β, IL-18and IL-33in serum of mice was higher than that in theother groups;(P<0.05)(4)After infected by H. pylori, gastric tissue of mice will produced more and moreCaspase-1;(5)As the eradication of H. pylori, the expression of NLRP3, ASC, Caspase-1, IL-1β,IL-18and IL-33reduced.Conclusion:(1) The expression of NLRP3inflammsomes and correlative molecular in the gastrictissue of infected mice increased suggest that NLRP3inflammsomes signal pathwaycould be activated by H. pylori infection;(2) There is a positive correlation between the inflammation of gastric tissue after theH. pylori infection and the expression of NLRP3inflammsomes and correlativemolecular(NLRP3, ASC, caspase-1, IL-1β, IL-18, IL-33).