| Objective To investigate the therapeutic effects of Balanophora involucrata extract on acetic gastric ulcer in rats,and to investigate the possible mechanism of action through the detection of proteins such as oxidative stress,inflammatory cytokines,epidermal growth factor and epidermal growth factor receptor.Methods Sixty male SD rats were randomly divided into sham-operated group,GU model group,omeprazole-positive group(3.6·mg-kg-1),and Balanophora involucrata low,medium and high dose treatment group(100,200 and 400·mg-kg-1).The GU model group was prepared by cautery with acetic acid injection,and the administration of the drug was started by gavage 1 day after surgery and continued for 10 day.The rats were continuously observed for general conditions such as mental,dietary and body weight.Samples were collected on the 5th and 10th day after administration to observe the general morphology of ulcers,and to measure and calculate the ulcer area and inhibition rate of each group;the histopathological changes of ulcers were observed by visual inspection combined with HE staining;the enzymatic assay was performed to detect the superoxide dismutase（SOD）activity,malondialdehyde（MDA）content and glutathione peroxidase（GSH-PX）activity in the gastric tissues of each group;the enzyme-linked immunosorbent assay（ELISA）was performed to detect the histopathological changes in the gastric tissues of each group.ELISA)to detect the contents of tumour necrosis factor-α（TNF-α）and interleukin-6（IL-6）in the gastric tissues of each group of rats;immunohistochemical staining（IHC）and protein immunoblotting（WB）to detect the localization and quantification of epidermal growth factor（EGF）and epidermal growth factor receptor（EGFR）proteins in the gastric tissues of each group of rats.Results 1.General:On the 5th day stage,compared with the sham-operated group,the rats in the GU model group were in a poorer mental state,with reduced diet and activity and reduced body weight（P<0.05）;compared with the GU model group,the rats in each treatment group had reduced all activities and increased body weight（P<0.05 or P<0.01）;on the 10th day stage,except for the sham-operated group,the rats in each group had significantly increased activity compared with before on the 10th day stage,except for the sham-operated group,the rats in all groups showed a significant increase in activity,smooth hair,and basic recovery of diet,and the weight of the GU model group decreased compared with that of the sham-operated group（P<0.05）;compared with the GU model group,the weight of the Balanophora involucrata treatment group increased（P<0.01 or P<0.001）.2.Ulcer area and ulcer inhibition rate:compared with the model group,the ulcer area of the Balanophora involucrata treatment group decreased on the 5th and 10th day stages（P<0.01）,and the ulcer inhibition rate decreased（P<0.01）.3.Pathological morphology:except for the sham-operated group,all groups showed round-like ulcerative lesions in the stomach lining,which were visible to the naked eye on the 5th and 10th day stages.HE staining showed that the gastric mucosa was defective at the ulcerative site,while the gastric mucosa of the sham-operated group was intact,with clear layers of the gastric wall and neatly arranged glands;the ulcerative site of the GU model group showed obvious inflammatory cell infiltration and disrupted glandular structure.The inflammatory cell infiltration was reduced,new granulation tissue was seen and the glandular structure tended to be normal in the Balanophora involucrata treatment group.4.Enzyme kit results:on the 5th and 10th day stages,compared with the sham-operated group,the gastric tissues of rats in the GU model group showed reduced SOD and GSH-PX activity（P<0.05 or P<0.01 or P<0.001）and increased MDA content（P<0.01 or P<0.001）.The SOD and GSH-PX activities were increased（P<0.05 or P<0.01 or P<0.001）and the MDA content was decreased（P<0.05）in the gastric tissues of the rats treated with Balanophora involucrata compared with the GU model group.5.ELISA results:On the 5th and 10th day stages,the TNF-αand IL-6 protein contents in the gastric tissues of the rats in the GU model group were increased（P<0.05 or P<0.01 or P<0.001）and decreased compared with the sham-operated group（P<0.05）.The results of IHC and WB assays:EGF and EGFR protein was expressed in the gastric mucosa,ulcer surface and inflammatory ooze when GU occurred.On the 5th and 10th day stages,EGF and EGFR protein expression increased in the GU model group compared with the sham-operated group（P<0.01 or P<0.001）;On the 10th day stages,,EGF and EGFR protein expression increased further in the Balanophora involucrata treatment group compared with the GU model group（P<0.05or P<0.01）.Conclusion Balanophora involucrata has therapeutic effects on acetic acid gastric ulcer in rats,and its mechanism may be related to reducing oxidative stress injury and inflammatory injury,increasing EGF and EGFR protein expression,and promoting the repair of damaged gastric mucosa. |
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