| Objective:By establishing a mouse model of Helicobacter pylori associated gastritis with damp-heat syndrome in the spleen and stomach,using the Qingre Qushi decoction to intervene in HAG mice,and observing the concentration and content of IL-1β、IL-18,the changes in m RNA content of micro RNA-204-5p 、 TLR4 、 NF-κB in the serum of mice before and after intervention,to explore the effects of Mieyou decoction on micro RNA-204-5p and TLR4/NF-κB signal pathway on inflammation of gastric mucosa epithelial cells in HAG mice.Methods:Twenty-eight SPF grade BALB/c mice were randomly divided into a blank group(8mice)and a model group(20 mice).The blank control group was given a normal feed diet and cloth type broth by gavage,while the model group was given a fat and sweet feed diet and Hp suspension by gavage,all of which were gavage once every other day,a total of 5 times.After 14 days of bacterial modeling and colonization,two mice were randomly selected from the blank control group and the modeling group to undergo rapid urease test and Warthin-Starry silver staining for gastric mucosa tissue.If the rapid urease test is positive,and clusters of short spiral black or dark brown substances are visible on the silver stained section under the microscope,it is considered that the modeling is successful.After successful modeling,18 mice in the modeling group were randomly divided into three groups: model control group(6 mice),standard quadruple group(6 mice),and Mieyou decoction group(6 mice),combined with the original blank control group(6 mice),a total of four groups.Both the blank control group and the model control group were given 0.2 ml of normal saline by gavage,while the standard quadruple group and Mieyou decoction group were given quadruple drug suspension and Mieyou decoction liquid0.2 ml by gavage,respectively.The four groups were gavage once a day for two consecutive weeks.Thirty minutes after the end of the last administration,the mice were killed,and serum and gastric mucosa tissue samples were taken for examination.HE staining was used to observe the pathological changes in the gastric mucosa of the mice,the content of IL-1β and IL-18 inflammatory factors in the serum was detected by Elisa method,and the m RNA content of micro RNA-204-5p,TLR4,and NF-κB in the gastric tissue was detected by RT-PCR.Results:1.General observation: Compared with the model control group,the mental state,food intake,and water consumption of mice in the standard quadruple group and the Mieyou decoction group improved after treatment,and their weight increased.The symptoms of mice in the standard quadruple group were significantly improved.2.Index detection: The HE staining results suggest that the gastric mucosal inflammation in the model control group mice is significantly improved,while the gastric mucosal inflammation in the standard quadruple group and Mieyou decoction group mice after treatment is significantly improved.The levels of serum IL-1β and IL-18 in model control group were significantly higher than those in blank control group by Elisa method(P<0.01);The expression of IL-1β and IL-18 in the standard tetralogy group and Mieyou decoction group was significantly lower than that in the model control group(P<0.01).RT-PCR detection showed that the m RNA content of micro RNA-204-5p in the gastric tissue of mice in the model control group decreased,while the m RNA content of TLR4 and NF-κB increased,with a statistically significant difference compared to the blank control group(P<0.01);The m RNA content of micro RNA-204-5p in the standard tetralogy group and Mieyou decoction group significantly increased,while the m RNA expression of TLR4 and NF-κB decreased,with a significant difference compared to the model control group(P<0.01);After treatment with standard quadruple drug or Mieyou decoction,the m RNA content of micro RNA-204-5p in gastric mucosa of both groups of mice increased,while the m RNA content of TLR4 decreased,with no statistically significant difference between the two groups(P>0.05).The content of NF-κB m RNA decreased,which was significantly higher in the standard quadruple group than in the Mieyou decoction group(P<0.05).Conclusion:The expression of micro RNA-204-5p in Hp infected gastric mucosa was significantly reduced,and increased expression of TLR4、NF-κB,thereby activating TLR4/NF-κB inflammatory signaling pathway,causing the inflammatory factor IL-1β、IL-18 and so on,so that gastric mucosa has severe inflammatory reaction.After treatment with Mieyou decoction,the expression of micro RNA-204-5p in gastric tissue increased,the expression of TLR4、NF-κB decreased,and the inflammation of gastric mucosa improved.It is speculated that Mieyou Decoction may promote the expression of micro RNA-204-5p gene in gastric mucosa,negatively regulate TLR4/NF-κB signaling pathway,inhibit cell apoptosis,thereby reducing gastric mucosal inflammation,and play a role in treating damp-heat syndrome of spleen and stomach in HAG. |
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