| Objective:In this study,Xinpuyin combined with omeprazole was used to intervene in a mouse model of Helicobacter pylori(H.pylori)-related gastritis.The efficacy of Xinpu decoction combined with omeprazole in eradicating H.pylori was explored.Intervention of Toll-like receptor-2(TLR-2)/Myeloid differentiation factor 88(MyD88)/Nuclear Factor kappa B(NF-κB)signaling pathway Effect on downstream inflammatory factors Tumor necrosis factor-α(TNF-α),Interleukin-1β(IL-1β),thereby clinically eradicating H.pylori and treating Helicobacter pylori-associated gastritis provides experimental basis as well as new methods and ideas.Methods:Fifty six-eight-week old SPF C57BL/6 mice(male and female half)were selected as subjects,and were labeled according to random number table and caged according to gender.They were divided into 5 groups:Control group(Normal group,Model group),Xinpuyin group,Xinpuyin+omeprazole(PPI)group,Western medicine triple group(amoxicillin+clarithromycin+omeprazole).Model group,Xinpuyin group,Xinpuyin plus PPI group,and Western medicine triple group for HP-related gastritis modeling:Inoculation of H.pylori liquid after 1 week of adaptive feeding,3 times of gavage(once every other day).After 4 weeks of colonization,random from each One male and one female were drawn from the group,and the rapid urease method and Giemsa staining method were used to test whether the modeling was successful.On the 42nd day,the drug-administered group was intragastrically administered with the corresponding concentration of the drug,and the control group was given normal saline twice a day for 2 weeks.All mice were sacrificed 4 weeks after the end of the gavage.The H.pylori eradication rate was detected by rapid urease method and Giemsa staining method,and Observing the gastric mucosa by HE staining under light microscope.Immunohistochemistry was used to detect the protein content of TLR2,MyD88 and NF-κB in gastric tissue was detected b.Western-Blot technique was used to detect the expression of TLR-2,MyD88,NF-κB,IκB kinase(IκK-α)and phosphorylated IκB-α(p-IκB-α)in gastric tissue;PCR technique was used to detect TLR-2,MyD88,NF-κB gene expression level in gastric tissue;enzyme-linked immunosorbent assay(ELISA)to detect serum TNF-α,IL-1β expression changes.Results:1.Symptoms of mouse model of H.pylori infection-related gastritis:After the Helicobacter pylori-associated gastritis mice were modeled,the external appearance of the mice in each group was observed,and it was found that the gastric inflammation symptoms of the model group mice were more obvious.After giving drug intervention,the symptoms of Xinpuyin group,Xinpuyin group plus PPI group,and Western medicine triple group improved significantly compared with the model group,and Xinpuyin group plus PPI group and Western medicine triple group were better than Xinpuyin group.2.H.pylori eradication:The H.pylori infection was negative in the normal group,and the H.pylori infection was positive in the model group.Compared with the model group,the eradication rate of H.pylori was increased in the Xinpuyin group,Xinpuyin plus PPI group,and the western medicine triplet group,and Xinpuyin plus PPI.The eradication rate of H.pylori in the three groups and the western medicine triplet group was higher than that in the Xinpuyin group.3.Observe inflammation of gastric mucosa by HE staining:No inflammation change was seen in the normal group,and obvious pathological changes in inflammation were seen in the model group.Compared with the model group,the pathological changes of inflammation were improved in the the pathological changes of inflammation Xinpuyin group,Xinpuyin plus PPI group,and the western medicine triplet group.The pathological changes of inflammation Xinpuyin plus PPI group and western medicine were superior to Xinpuyin plus PPI group.Triad group and Xinpu drink group.4.IHC technology test results:Compared with normal values,the expressions of TLR2,MyD88,and NF-κB in gastric mucosa of the model group were significantly higher than those in the normal group(P<0.01);compared with the model group,the expression in gastric tissues of mice in the Xinpuyin group,Xinpuyin+PPI group,Western medicine triplet group was significantly reduced(P<0.01).Compared with Xinpuyin group,the expression in gastric tissues of mice in Xinpuyin+PPI group and Western medicine triplet group was significantly reduced.(P<0.05).the expression levels of TLR2,MyD88,and NF-κB in the Xinpuyin+PPI group were lower than those in the western medicine triple group(P>0.05).5.WesternBlot technology test results:Compared with the normal group,the expression levels of MyD88,NF-κB,IκK-α,and p-IκB-α in the gastric mucosa of the model group were significantly increased(P<0.01);compared with the model group,Xinpuyin Group,Xinpuyin+PPI group and western medicine triplet group significantly reduced the expression of gastric tissue in mice(P<0.01);compared with Xinpuyin group,Xinpuyin+PPI group and western medicine triplet group mice gastric tissue expression The expression level was significantly decreased(P<0.05);the expression level in Xinpuyin+PPI group was lower than that in the Western medicine triplet group,but there was no significant difference(P>0.05).6.qPCR technology test results:Compared with normal values,the expression levels of TLR2,MyD88,and NF-κB mRNA in gastric mucosa of the model group were significantly higher than those in the normal group(P<0.01);compared with the model group,the expression of mRNA in gastric tissues of mice in the Xinpuyin group,Xinpuyin+PPI group and western medicine triplet group was significantly reduced(P<0.01);compared with Xinpuyin group,mRNA expression in gastric mucosa of mice in Xinpuyin+PPI group and western medicine triplet group was significantly reduced(P<0.05),and the expression of mRNA in Xinpuyin+PPI group was lower than that in the western medicine triad group,but there was no significant difference(P>0.05).7.Elisa technology test results:Compared with the normal group,the expression levels of TNF-α and IL-1β in the serum of the model group mice were significantly increased(P<0.01);compared with the model group,the expression levels in the serum of the mice in the Xinpuyin group,Xinpuyin + PPI group and the western medicine triplet group were significantly reduced(P<0.01);compared with the Xinpuyin group,the expressions in the serum of the Xinpuyin+PPI group and the western medicine triplet group were significantly reduced(P<0.05).The expression levels of TNF-α and IL-1β in the serum of Xinpuyin+PPI group were lower than those in the western medicine triplet group,but there was no significant difference(P>0.05).Conclusion:1.The mechanism of Helicobacter pylori-associated gastritis may be related to the activation of TLR-2/MyD88/NF-κB signaling pathway and the release of downstream inflammatory factors TNF-α and IL-1β.2.Xinpuyin has the effect of eradicating H.pylori.Combined with PPI,the effect is betterand comparable to Western medicine triple.3.Xinpuyin and Xinpuyin combines with PPI may exert anti-inflammatory effects by inhibiting the activation of TLR-2/MyD88/NF-κB signaling pathway and the release of TNF-α and IL-1β.While Xinpuyin combines with PPI,the effect is more significant. |
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