| Objective:To investigate the Repairing Effect and mechanism of TCM Combined Therapy of harmonizing stomach,regulating the vital energy flow and Shou-Gong(gecko)treatment on the repairing of gastric mucosa of chronic atrophic gastritis model rats from different aspects of histomorphology(staining sections of HE and CD34),anti-oxidative damage(SOD、MDA),cell proliferation and apoptosis(P53 and PCNA)and signal pathway(Wnt pathway).Methods:72 healthy SPF grade pure breed SD male rats were selected and fed in a barrier environment.After adaptive feeding of one week,the chronic atrophic gastritis model was made by feeding deoxycholate citrate lactose saccharose(agar)water solution with ethanol and hunger& gorge method.The model rats were grouped as blank control group(K),model group(M),TCM group(Z),Weifuchun group(W),folic acid group(Y)in 10 rats per group.Intragastric administration was given from the third day after the success of modeling,and the blood samples were sampled after 28 days of drug intervention.The pathological changes of gastric mucosa weremonitored by HE staining.The CD34 immunohistochemical staining of gastric mucosa was introduced to detect the microvessel count and density.The ELISA method was used to evaluate the contents of serum gastrin(GAS),superoxide dismutase(SOD)/malondialdehyde(MDA),serum somatostatin(SS)/serum prostaglandin E2(PGE2).The immunohistochemistry method was applied to evaluate the gene expression level of gastric mucosal P53 and PCNA,and the protein expression level of GSK-3 beta,beta-catenin,cyclin D1 and COX-2.Results:1.Effects on the histomorphology of chronic atrophic gastritis model ratsHE staining showed the erosion and defects of the gastric mucosa of the model group rats.Some severe cases have gastric ulcer lesions,with mucosal thinning.The glands are disorganized and in different sizes,and the number also reduced for more than 2/3,with some intestinal metaplasia and atypical hyperplasia lesions.Gastric mucosa erosion and defects can be occasionally seen in the TCM group,with normal mucosal thickness and no gastric ulcer lesions.The glands are orderly arranged,with no significant reduction in number(less than 1/3),no intestinal metaplasia and atypical hyperplasia lesions.There were few erosion and defects in gastric mucosa of the Weifuchun group,with occasional gastric ulcer lesions.The thickness of mucosa layer was increased,and the arrangement of the glands was slightly disordered.The number of the glands was reduced for about 1/2.Occasional intestinal metaplasia and atypical hyperplasia can be spotted.In the folic acid group,there were few erosions and defects in the gastric mucosa,and there were occasional gastric lesions.The thickness of the mucosal layer was increased and the arrangement of the glands was slightly disordered in a decreased number for about 1/2.Occasional intestinal metaplasia and atypical hyperplasia can be observed.The microvessel count(MVC)and density(MVD)of gastric mucosa detected by CD34 immunohistochemical staining showed that: the MVD of the model group was significantly lower(P<0.01)comparing to that of the blank control group;that of the treatment group was significantly increased(P<0.01)comparing to that of the model group;immunohistochemical staining showed the a different level of hyperemia responses of the blank group,while the model group had a hyperplasia of gastric mucous epithelium and glands cell,and a reduction of the interstitial blood vessels of mucosa and submucosa.After the treatment of TCM,the microvessel density of gastric mucosa was increased towards the normal level.The statistical analysis showed that there was no difference between the treatment group and the blank control group(P<0.05).2.The levels of GAS,SS,PGE2,SOD and MDA in the serum of chronic atrophic gastritis rats.The serum gastrin(GAS)levels of the model group rats were significantly lower(P < 0.01)comparing to that of the blank control group.The gastrin(GAS)levels of the TCM group,the folic acid group and the Weifuchun group were significantly increased(P<0.01)comparing to that of the model group.Compared with the Weifuchun group,the serum gastrin(GAS)level of the TCM group increased(P < 0.05).Statistically,there is no significantdifference in serum gastrin(GAS)(P > 0.05)between the TCM group and the folic acid group.Compared with the blank control group,the serum somatostatin(SS)and prostaglandin(PGE2)of the model group decreased significantly(P < 0.01).The serum somatostatin(SS)and prostaglandin(PGE2)of the TCM group significantly increased(P<0.01)comparing to that of the model group.The serum somatostatin(SS)and prostaglandin(PGE2)level of the folic acid group and Weifuchun group were increased(P >0.05)after treatment.No statistical difference between the TCM group and the folic acid group,Weifuchun group in the change of serum somatostatin(SS)and prostaglandin(PGE2)level.Compared with the blank control group,the serum superoxide dismutase(SOD)activity decreased and the malondialdehyde(MDA)value increased(P<0.05)in the model group.Compared with the model group,the SOD activity of TCM group was significantly increased,and the MDA decreased significantly(P<0.01).A similar change is also observed for the folic acid group and Weifuchun group.No statistical difference between the TCM group and the folic acid group,Weifuchun group in the change of SOD and MDA level.3.Effects on the gene expression of PCNA and P53 in rat gastric mucosaCompared with the blank control group,the level of P53 in the gastric mucosa of the model group was significantly higher than that of the normal group(P<0.01)and the pathological changes of gastric mucosa is evident.The P53 index of different groups was lowed but the histopathological changes of gastric mucosa were significant.The difference between the TCM group and model group(and Weifuchungroup)was significant(p<0.01).No big difference between the TCM group and folic acid group(P > 0.05).Compared with the control group,the level of PCNA in the gastric mucosa of the model group was significantly higher than that of the normal group(P<0.01)and the pathological changes of gastric mucosa is evident.The PCNA index of different treatment groups was lowered but the histopathological changes of gastric mucosa were significant.Chinese medicine treatment group and model group were significantly different(p<0.01);Chinese medicine treatment group and the stomach of the spring group was significantly different(P< 0.01).The difference between the TCM group and model group(and Weifuchun group)was significant(p<0.01).4.Effects on the gastric mucosa Wnt signal pathway in ratsCompared with the blank control group,the expression of GSK-3β in the TCM group,the folic acid group and the Weifuchun group was increased;The difference was statistically meaningful(P<0.01).When compared in pairs among the three groups,the TCM group was better the Weifuchun group(P<0.01)and the folic acid group(P<0.01).Statistically,there was no difference between the Weifuchun group and the folic acid group(P > 0.05).Compared with the blank control group,the expression of β-catenin in the model group was increased;The difference was statistically meaningful(P<0.01).The expression of GSK-3β in the TCM group,the folic acid group and the Weifuchun group was decreased;the difference was statistically meaningful.When compared in pairs among the three groups,the TCM group was better the Weifuchun group(P<0.05).Statistically,it is not meaningfulto compare the folic acid group and the TCM group(P > 0.05)or the Weifuchun group.Compared with the blank control group,the expression of cyclin D1 in the model group was increased;the difference was statistically meaningful(P<0.01).The expression of GSK-3β in the TCM group,the folic acid group and the Weifuchun group was decreased;the difference was statistically meaningful(P<0.05).When compared in pairs among the three groups,the TCM group was better the Weifuchun group(P<0.01).Compared with the blank control group,the expression of COX-2in the model group was increased;the difference was statistically meaningful(P<0.01).The expression of GSK-3β in the TCM group,the folic acid group and the Weifuchun group was decreased;the difference was statistically meaningful(P<0.01).When compared in pairs among the three groups,the TCM group was better the Weifuchun group(P<0.01)and the folic acid group(P<0.01).Conclusions:TCM Combined Therapy of harmonizing stomach,regulating the vital energy flow and Shou-Gong(gecko)treatment has better effects in repairing the ventriculi phthisis,improving neovascularization and promoting angiogenesis of gastric mucosa than that of Weifuchun and folic acid treatments.It can increase the levels of gastrin,somatostatin and prostaglandin E2,reduce MDA,suppress inflammation,repair damaged gastric mucosa,and increase SOD activity.Such effects can be related to the content of gastrin,somatostatin,prostaglandin E2,SOD and MDA in gastric mucosa.It can prevent the overexpression of P53 and PCNA in gastric mucosa,thus preventing the transformation of chronic atrophic gastritis into gastric cancer,indicating its advantages in treating the chronic atrophic gastritis.Its repairing effect of gastric mucosa can be related to the suppression of the overexpression of P53 and PCNA.It can improve the expression of GSK-3β,cyclin D1 and COX-2,reduce the expression of β-catenin of gastric mucosa.The gastric mucosal repair mechanism is likely to improve the growth factor in gastric mucosa,activate the Wnt pathway to reverse the gastric mucosa atrophy.In summary,the TCM Combined Therapy of harmonizing stomach,regulating the vital energy flow and Shou-Gong(gecko)treatment has a better effect on the repair of gastric mucosa than that of Weifuchun method or folic acid method.It can effectively suppress the oxidative stress reaction and improve the repair ability of gastric mucosa.This thesis provides some solid experimental data for the treatment of chronic atrophic gastritis with TCM combined therapy of harmonizing stomach,regulating the vital energy flow and Shou-Gong(gecko)treatment. |
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