| Objective: This study is based on Traditional Chinese Medicine(TCM)theory "spleen governing muscles",trying connotate its "muscles" to the stomach,which is a internal muscular organ.We established the rat GU model induced by glacial acetic acid,and combined with our previous research,to discuss the effect of Jiuwei Yiwei Mixture(JYM)for gastric mucosal repair and MEK/ERK signal pathway in rats with gastric ulcer.Methods: 66 healthy specific pathogen free(SPF)rank SpragueDawley male rats were chosen,raised in SPF environment barrier.Adaptive feeding for 7 days,12 rats were randomly selected as the pretreatment group(group Y),give JYM by intragastric administration for 7 days,while the remaining groups were given an equal volume of physiological saline.7 days later,another 12 rats were randomly selected as control group(group K),except for group K,the rat GU model induced by glacial acetic acid were established according to Okabe’s method.3 days after duplicating model,2 rats were randomly selected from group K,group Y and the rats without grouping,to evaluate whether GU were successfully copied on rats.The remaining 40 rats were randomly divided into model group(group M),Jiuwei Yiwei Mixture group(group J),JianpiYiqi group(group P),hydrotalcite group(group L),add group K and group Y,there were 6 groups in total with 10 rats in each group.3 days after the model making operation,the rats were given intragastric administration,drug intervention lasted for 28 days.After the final administration,all the rats were killed by fasting for 24 h,samples of blood and stomach tissue were collected.HE staining was used to observe the pathological changes of the GU marginal tissues in rats,the absent width of gastric mucous and the thickness of regenerated mucous were measured.Enzyme linked immunosorbent assay(ELISA)method was used to measure the content of SDH and TFF3 in serum and the content of PCNA and TGF-α in gastric mucosa homogenate.The expression levels of EGFR,ZO-1 and Occludin in gastric tissue were measured by immunohistochemistry;WesternBlot was used to detect the CyclinD1,p-MEK and p-ERK in the GU marginal tissues;Real-Time PCR(RT-PCR)detection of the expression of MEKmRNA and ERKmRNA in the GU marginal tissue.Results:(1)Effect of the absent width of gastric mucous and the thickness of regenerated mucous: The gastric mucosa was normal in group K,was thinned and accompanied by significant morphological and pathological changes in group M,while were basically normal in group J,group P,group L and group Y.Compared with the group M,the absent width of gastric mucous in the rest groups were significantly decreased(P<0.01)and the thickness of regenerated mucous were significantly increased(P<0.01).Compared with the group L,both the absent width of gastric mucous and the thickness of regenerated mucous were no significant difference(P>0.05)in group J and group P.The absent width of gastric mucous in group Y were significantly lower than in group J,group P and group L(P<0.01),while the thickness of regenerated mucous were significantly higher(P<0.01).(2)Effect of gastric mucosa cell proliferation: Compared with the group K,the expression of PCNA in gastric tissue of group M was increased(P<0.05),while in group J,group P,group L and group Y were significantly increased(P<0.01).Compared with the group M,the expression of PCNA in gastric tissue of the rest groups were increased(P < 0.05 or P < 0.01).There were no significantly differences between group J,group P and group L(P>0.05).Compared with the group Y,the expression of PCNA in gastric tissue of group P was decreased(P<0.05),while no significantly differences were showed in group J and group L(P>0.05).Compared with the group K,the expression of CyclinD1 in the rest groups were significantly increased(P<0.01).Compared with the group M,the expression of CyclinD1 in group J,group P,group L and group Y were increased(P<0.01).The expression of CyclinD1 in group L was lower than group J and group Y(P<0.01),while higher than group P(P<0.05).Compared with the group Y,the expression of CyclinD1 in group P was decreased(P < 0.05),while no significantly differences were showed in group J(P>0.05).(3)Effect of the expression of gastric mucosal repair related gastrointestinal peptides: (1)Compared with the group K,the expression of TGF-α and EGFR of the rest groups were significantly increased(P<0.01).Compared with the group M,the expression of TGF-α and EGFR in group J,group P,group L and group Y were also significantly increased(P<0.01).Compared with the group L,the expression of TGF-α and EGFR showed no significantly differences in group J and group P(P>0.05).Compared with the group Y,the expression of TGF-α and EGFR of group J,group P and group L were significantly decreased(P<0.01).(2) Compared with the group K,the expression of TFF3 in the rest groups were significantly increased(P<0.01).Compared with the group M,the expression of TFF3 in group J,group P,group L and group Y were significantly increased(P<0.01).There were no significantly differences between group J,group P,group L and group Y(P>0.05).(4)Effect of the expression of tight junction proteins in gastric mucosa: Compared with the group K,the expression of ZO-1 and Occludin in gastric tissue in group M,group J and group P were significantly decreased(P<0.01),while it showed no differences in group L and group Y(P > 0.05).There were no significantly differences between group L and group Y(P>0.05),and the expression of ZO-1 and Occludin between group P and group J also showed no differences(P>0.05).The expression of ZO-1 and Occludin in group L and group Y were higher than in group P and group J(P<0.05).(5)Effect of mitochondrial function related markers SDH expression: Compared with the group K,the expression of SDH showed no significantly differences between group J,group P and group Y(P>0.05).SDH expression in group M and group L were decreased lower than the other groups(P<0.01).Compared with the group M,the expression of SDH were increased in group J,group P,group L and group Y(P<0.01).No significantly differences were showed between group J,group P and group Y(P>0.05),and the expression of SDH in those 3 group were higher than group L(P<0.01).(6)Effect on MEK/ERK expression in gastric mucosa: Compared with the group K,the expression of MEK/actin and ERK/actin decreased,while p-MEK/actin and p-ERK/actin were lower in group M(P<0.05).Compared with the group M,the expression of MEK/actin and ERK/actin increased,p-MEK/actin and p-ERK/actin were higher in group J,group P,group L(P < 0.05).Compared with group L,p-MEK/actin was lower than group J while higher than group P and group Y(P<0.01);Compared with group Y,p-MEK/actin and MEK/actin were lower than group J while higher than group P(P<0.01);there were no significantly differences between group L and group Y in p-ERK/actin(P>0.05),while both two groups were higher than group J and group P(P<0.01);Compared with the group L,MEK/actin was lower than group J while higher than group P(P<0.01),and no significantly differences between group Y(P>0.05);Compared with the group L,ERK/actin was lower than group J and group Y,while higher than group P(P<0.01);Compared with the group Y,ERK/actin was higher than group P(P<0.01),and showed no significantly differences with group J(P>0.05).Compared with the group K,the amplification of p-MEK mRNA and p-ERK mRNA was decreased in group M(P<0.05).Compared with the group M,the amplification of p-MEK mRNA and p-ERK mRNA were increased in group J,group P,group L and group Y(P<0.05).Compared with the group L,no significantly differences were showed in p-MEK mRNA and p-ERK mRNA between group J,group P and group Y(P>0.05).Compared with the group Y,no significantly differences were showed in p-MEK mRNA and p-ERK mRNA in group J and group P(P>0.05).Conclusions: 1.JYM can be used to repair the pathological damage of gastric mucosa induced by glacial acetic acid,increase the thickness of regenerated mucosa and reduce the absent width of gastric mucous,improve the histological maturity of regenerated mucosa.2.JYM can increase the expression of PCNA and CyclinD1 in gastric mucosa,which might be the mechanism of promoting the proliferation of mucosal cells and repairing the mucosal defects.3.JYM can increase the expression of TGF-α and TFF3,which play an important role in the maintenance of gastrointestinal mucosal structure and injury repair process,activated EGFR as well.It’s suggested that the increase expression of TGF-α and TFF might be related to participate in the regulation of cell proliferation and migration in the process of gastric mucosal repair.4.JYM can enhance the expression of tight junction proteins ZO-1 and Occludin of mucosal epithelium,preadministration intervention can further enhance this trendence,these suggested that the tight junction structure can be strengthened,which indicate enhanced mucosal mechanical barrier function.JYM showed some advantages not only in gastric mucosal repair,but also in gastric mucosal injury prevention.5.JYM can increase the expression of SDH,a function marker of mitochondrial,show that strengthening the spleen-Qi therapy can enhance mitochondriain function in gastric tissue,suggesting that those kind of TCM prescriptions might contribute to GU repair.6.JYM can up-regulate MEK and ERK expression,the mechanism of JYM’s promoting gastric mucosal repair might improve the utility of gastrointestinal peptides,activate MEK/ERK pathway,promote the process of gastric mucosal cell proliferation and migration,as a result,gastric mucosal were repaired and reconstructed.7.The comprehensive index of groups given JYM is better than that given only invigorating Spleen and replenishing Qi,which is JYM’s decomposed recipes.It indicates that although JYM’s effect on GU prevention and treatment is centered around TCM’s Spleen,its echanism is still characteried by multi targets and complex.This is in line with various factors involved in the pathogenesis of GU.This study suggests that the JYM can participate the treatment and prevention of GU effectively,its mechanism might involve several aspects:improve the utility of gastrointestinal peptides,activate MEK/ERK pathway,promote gastric mucosal cell proliferation and migration,enhanced mucosal mechanical barrier function,et al.In same level,this study verify the connotation of“spleen governing muscles”is included the spleen governing growth and repair of visceral muscles.This might provide some new ideas for the prevention and treatment of GU. |
PDF Full Text Request