| Objective: To observe the effect of Yufuning Granule on gastric tissue morphology,serum TNF-alpha,IL-6,IL-8 and expression of TLR4,I-kappa B and NF-kappa B proteins in rats with chronic non-atrophic gastritis by free drinking ammonia water combined with starvation and satiety disorder,and to explore the mechanism of Xuefuning Granule on the signal transduction pathway of NF-kappa B in rats with chronic non-atrophic gastritis.Methods: Sixty Wistar rats,weighing 180-220 g,SPF grade,half male and half female,were randomly divided into normal group,model group,control group and treatment group,10 rats in low,medium and high dose groups,10 rats in each group.Normal control group was fed with standard feed.Model group,control group and treatment group were fed with 0.02% ammonia water every day freely.At the same time,they were fed with starvation and satiety disorder(1).The chronic non-atrophic gastritis model was duplicated by fasting for two days.After 90 days,the model was given treatment.The normal group continued to be fed with standard rat feed,and normal saline was given daily(0.8ml/100g).The model group was given normal saline(0.8ml/100g)daily,while the control group was given omeprazole(6.6g/kg),while the treatment group was given low,medium and high doses of Yaofuning Granules(1.7g/kg,5g/kg,15g/kg),10 rats in each group.At the same time,the model group,the control group and the treatment group continued to drink 0.02% ammonia water freely.After 14 days of continuous administration,the gastric tissues were taken for HE staining to observe the pathological changes of gastric mucosa.The expression of TNF-a,IL-6 and IL-8 in serum was determined by enzyme-linked immunosorbent assay(ELISA).The expression of TLR4,I-kappa B and NF-kappa B in gastric tissues was detected by immunohistochemical staining.Polymerase chain reaction(RT-PCR)was used to detect the expression of I-kappa B protein in gastric tissue at transcriptional level;Western blot was used to detect the protein content of TLR4,I-kappa B and NF-kappa B in gastric tissue of rats.Results:1.Effect of Xiefuning Granule on Pathological Changes of Gastric Mucosa in Rats with Chronic Non-atrophic GastritisAfter 90 days of modeling,the general state of rats,including hair,diet,mental state,defecation and weight,was significantly different between the model group and the normal group(p< 0.05).After intervention,the weight of the high-dose group increased faster than that of the normal group(p> 0.05).The weight of the medium-dose group increased faster than that of the normal group(p> 0.05).The weight of the low-dose group increased slowly,with statistical significance compared with that of the normal group(p< 0.05).There was no significant difference between the dose group and the control group(p> 0.05).there was no significant difference between the middle dose group and the control group(p > 0.05).there was significant difference between the low dose group and the control group(p<0.05).The results of HE staining showed that the epithelium of gastric mucosa in model group was incomplete,superficial epithelium necrosis and exfoliation,gastric mucosa and submucosa showed focal or diffuse distribution,gastric mucosa congestion and edema,even erosion and hemorrhage,lymphocyte and plasma cell infiltration in lamina propria,but there was no obvious gland destruction.Compared with the normal group,the difference was significant(p< 0.05),suggesting that the model was successful.The gastric mucosal epithelium of model group was incomplete,necrosis and exfoliation of epithelial cells,lymphocyte and plasma cell infiltration were the most obvious,and there was significant difference compared with normal group(p< 0.05).The pathological changes of gastric mucosa of rats in high,middle,low dose group and control group were improved in varying degrees,and the difference was statistically significant compared with model group(p< 0.05).There was no significant difference in the gastric mucosa between the middle dose group and the control group(p< 0.05),but there was no significant difference in the gastric mucosa between the low dose group and the control group(p> 0.05).2.Effect of Xiefuning Granule on Serum TNF-alpha,IL-6 and IL-8 in Rats with Chronic Non-atrophic GastritisTNF-alpha model group compared with the normal group,the difference was significant,with statistical significance(p<0.05),high,medium,low dose group and control group were lower than the model group(p<0.05),with statistical significance;high and medium dose group rats TNF-alpha slightly better than the control group,with no significant difference(p> 0.05),and low dose group rats compared with the control group(p< 0.05),there was significant difference(p < 0.05).Statistical significance.IL-6 model group compared with the normal group,the difference was significant,with statistical significance(p< 0.01),the content of IL-8 is shown in table 4,model group compared with the normal group,the difference was significant,with statistical significance(p< 0.01).IL-6 and IL-8 in high,medium and low dose groups and control groups were lower than those in model group(p< 0.05),with statistical significance;IL-6in high and medium dose groups was significantly better than that in control group(p<0.05),with statistical significance;compared with control group,IL-6 in low dose group had no statistical significance(p> 0.05);and IL-6 in low dose group was higher than that in high and medium dose groups(p< 0.05),with statistical significance.3.Effect of Xiefuning Granule on TLR4,I-kappa B and NF-kappa B in Gastric Tissue of Rats with Chronic Non-atrophic GastritisThe results of immunofluorescence showed that there were significant differences between TLR4 and NF-kappa B model group and normal group(p< 0.05),suggesting that the levels of TLR4 and NF-kappa B in serum of rats in chronic non-atrophic gastritis model group increased.TLR4 in high and middle dose group was lower than that in model group(p< 0.01),with statistical significance;TLR4 in low dose group was lower than that in model group(p< 0.05),with statistical significance;TLR4 in high and middle dose group was not significantly different from that in control group(p> 0.05),without statistical significance;TLR4 in low dose group was significantly lower than that in control group(p< 0.05);NF-kappa B in high,medium and low dose group was lower than that in control group(p< 0.05).The model group(p< 0.05)had statistical significance;there was no significant difference between the high and middle dose groups(p> 0.05)and the control group(p< 0.05);there was statistical significance between the low dose group and the control group(p < 0.05).The results of immunofluorescence showed that there was a significant difference between the I-kappa B model group and the normal group(p< 0.05).The high,middle and low dose groups were higher than the model group(p< 0.05),with statistical significance;there was no significant difference between the high and middle dose groups and the control group(p> 0.05),and there was no statistical significance between the low dose group and the control group(p< 0.05).4.Effect of Xiefuning Granule on I-kappa B Protein mRNA in Gastric Tissue of Rats with Chronic Non-atrophic GastritisRT-PCR results showed that the level of I-kappa B in gastric tissue of rats was significantly different from that of normal group(p< 0.05),suggesting that the serum I-kappa B level in chronic non-atrophic gastritis model group was decreased.The high,middle and low dose groups were higher than the model group(p< 0.05),with statistical significance;the high and middle dose groups had significant difference(p< 0.05)compared with the control group;the low dose group had no statistical significance compared with the control group(p> 0.05);the low dose group was lower than the high and middle dose groups(p< 0.05),with statistical significance.5.Effect of Xiefuning Granule on the Protein Contents of TLR4,I-kappa B and NF-kappa B in Gastric Tissue of Rats with Chronic Non-atrophic GastritisTLR4,I-kappa B and NF-kappa B model group compared with the normal group,the difference was significant,with statistical significance(p< 0.05).TLR4 and NF-kappa B in high,medium and low dose groups were lower than those in model group(p< 0.05),with statistical significance;TLR4 in high and medium dose groups was significantly different from that in control group(p< 0.05),with statistical significance;there was no significant difference between high and medium dose groups(p> 0.05),and there was no statistical significance;there was no significant difference between low dose group and control group(p< 0.05).Statistical significance;I-kappa B high,medium and low dose group and control group were higher than the model group(p< 0.05),with statistical significance;I-kappa B in high and medium dose group had no significant difference compared with the control group(p> 0.05),without statistical significance;rats in low dose group had statistical significance compared with the control group(p< 0.05).Conclusion:1.Xiefuning Granule can improve the general state of chronic non-atrophic gastritis rats,including hair,diet,mental state,defecation and weight.2.Xiefuning Granule can reduce the index of gastric mucosal damage in rats with chronic non-atrophic gastritis and the degree of pathological damage of gastric mucosa.3.The expression of TNF-alpha,IL-6 and IL-8 in serum of rats with chronic non-atrophic gastritis could be reduced by Xiefuning Granule.4 From the molecular level and protein quantitative analysis,it can be confirmed that Huiweining Granule can reduce the content of TLR4 and NF-kappa B and enhance the expression of I-kappa B in rats with chronic non-atrophic gastritis.5.By reducing the expression of TNF-a,IL-6,IL-8,inhibiting inflammatory factors,reducing inflammation,and inhibiting the levels of TLR4 and NF-kappa B in NF-kappa B signaling pathway,the granule can enhance the level of inhibitory activator I-kappa B.6.The granule can regulate the signal transduction system of NF-kappa B in rats with chronic non-atrophic gastritis.It can affect the binding of NF-kappa B to the nucleus and target gene.The granule may be related to the inhibition of the signal transduction system of NF-kappa B in rats with chronic non-atrophic gastritis. |
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