| ObjectiveThis experiment aims to explore a more efficient and sucessful animal model of exercise-induced stress ulcer(ESU),and through the detection of gastric mucosal oxidative stress products,cytokines and JAK2/STAT3 signal-related protein expression,explore the protective mechanism of Banxia Xiexin Decoction(BXD)on the gastric mucosa of rats with exercise stress ulcer.MethodsExperiment one:40 rats were randomly divided into blank group(n=8),one-time exhaustion group(n=8),incremental training exhaustion group(n=12)and daily exhaustion group(n=12).Except for the blank group,all groups were modeled by exhaustive swimming with 3%body weight.The rats in the one-time exhaustion group only performed once weightbearing exhausted swimming,and the incremental training exhaustion group increased the swimming time every week to increase the amount of exercise.The rats in the exhaustion group swim to exhaustion every day.The animal behavior analysis was completed through the open-field test before the last exhaustive swimming.After the last exhaustive swimming,the samples were collected to observe the anatomical morphology and pathological changes of the gastric mucosa of each group,calculate the ulcer index,and detect the SOD and MDA in the rat serum content.Experiment two:50 healthy SD rats were randomly divided into blank group(n=10),model group(n=10),traditional Chinese medicine group(n=10),western medicine group(n=10),JAK2/STAT3 pathway inhibitor(AG490)group(n=10).The model was created by swimming exhaustion with a one-time weight bearing 3%of the body weight.Drug intervention was started one week before modeling.The traditional Chinese medicine group was given BXD and the western medicine group was given omeprazole.The AG490 group started to model after 30 minutes of intraperitoneal injection of AG490.Observe the anatomical morphology and pathological changes of the gastric mucosa of each group,calculate the ulcer index,and determine the expression of TNF-α,IL-1β,and IL-6 in the gastric mucosal tissue by enzyme-linked immunosorbent assay.Western Blot was used to detect the protein expression of JAK2/STAT3 signaling pathway in gastric mucosa.Results1.Exploration of Modeling1.1 Open-field test:The total distance of rats in the one-time exhaustion group and incremental training group is not significantly different from that of the blank group.P>0.05.The total distance of the rats in the daily exhaustion group is significantly less than that of the blank group,P<0.05.The average speed and activity of the rats in the daily exhaustion group were significantly lower than that of the blank group.P<0.05.Compared with the blank group,the other groups had no significant difference,P>0.05.Judging from the activity trajectory diagram.there is no significant difference in the activity of the one-time exhaustion group and the incremental training exhaustion group compared with the blank group.The activity of the rats in the daily exhaustion group is poor and lacks the desire to explore.The weight gain of rats in the daily exhaustion group and the incremental training exhaustion group was slow,and the weight growth rate was significantly lower than that of the blank group,whose difference was statistically significant,P<0.05.The water intake and food intake of the one-time exhaustion group were equivalent to those of the control group,P>0.05;while the incremental training exhaustion group and the daily exhaustion group had significantly less food intake than the control group,and the amount of drinking water was significantly higher than that of the blank group,P<0.05,in which the difference in the daily exhaustion group was more obvious.1.2 Ulcer index:Compared with the blank group,the gastric ulcer index of the rats in the one-time exhaustion group,the incremental training exhaustion group,and the daily exhaustion group was significantly increased.P<0.05,in which the one-time exhaustion group was the highest,followed by the daily exhaustion group and the incremental training exhaustion group.1.3 Pathological observation:All three modeling methods can cause stress gastric ulcer.The mucosal structure of the blank group is intact without defects,the glands are arranged neatly,and there is no obvious cell or tissue proliferation.The gastric mucosa of rats in the one-time exhaustion group showed defective ulcers on the gastric mucosal epithelium,a large amount of red-stained necrotic material that lost normal tissue structure,and degenerated yellow or tan necrotic tissue,as well as exfoliated epithelial cells,and the ulcer reaches deep to the muscularis mucosa,a large number of red blood cells and inflammatory cells infiltrate and infiltrate.In the daily exhaustion group,the gastric mucosal epithelial interstitial edema was obvious.The cells secreted a large amount of mucus,and the gastric pits were deep Ushaped,and a large number of red blood cells and inflammatory cells were gathered locally,or the gastric mucosal epithelium was arranged irregularly,and even worse erosion and loss of the epithelial tissue of the gastric mucosa occur,but it does not exceed the lamina propria.The gastric mucosal epithelial glands of the rats in the incremental training exhaustion group are arranged disorderly,partially absent or eroded,the epithelial cells secrete more mucus,cell edema,a little vacuole-like change,and red blood cell aggregation can be seen in the gap.1.4 Serum SOD and MDA content test:Compared with the blank group,the SOD content in the serum of the rats in the one-time exhaustion group,the incremental training exhaustion group,and the daily exhaustion group were significantly increased.P<0.05.and the expression level of the one-time exhaustion group and the daily exhaustion group is the highest.Compared with the blank group,the MDA content of the one-time exhaustion group and the incremental training exhaustion group was close to the expression level of the blank group,and the difference was not statistically significant.P>0.05.The content of the rats in the daily exhaustion group was significantly higher than that of the blank group,P<0.05.1.5 Survival rate and model-building success rate:The survival rate and model-building success rate of the one-time exhaustion group and the incremental training exhaustion group are significantly better than the daily exhaustion group.2.Research on the protective mechanism of BXD on exercise-induced stress ulcer2.1 Expression of TNF-α,IL-1β,and IL-6 in gastric mucosa:The expression of IL-1β,IL-6,and TNF-α in gastric mucosa of the blank group was significantly lower than that of the model group,and the difference was statistically significant(P<0.05).The expression levels of inflammatory factors IL-1{,IL-6 and TNF-α in the traditional Chinese medicine group and the western medicine group were also significantly lower than those in the model group,and the difference was statistically significant(P<0.05);IL-1β,IL in the traditional Chinese medicine group-6.There was no significant difference in the expression of TNF-αcompared with the western medicine group(P>0.05).2.2 Ulcer index:The UI of the gastric mucosa of the blank group was significantly lower than that of the model group,the difference was statistically significant(P<0.05).The UI of the traditional Chinese medicine group,the western medicine group and the AG490 group was significantly lower than that of the model group,and the difference was statistically significant(P<0.05).There was no significant difference in UI between the traditional Chinese medicine group and the western medicine group(P>0.05).2.3 Anatomical morphology and pathological observation of the gastric mucosa:The gastric mucosa of the model group is obviously congested,red and swollen,with a large number of linear and flaky bleeding spots;the western medicine group,the traditional Chinese medicine group and the AG490 gastric mucosa have a little congestion and a mildly bleeding.In the blank group,the mucosal structure was intact and there was no defect,the epithelial layer,lamina propria,and muscular layer were distinct,the glands were arranged neatly,and there was no obvious cell or tissue hyperplasia.The epithelial layer of the gastric mucosa of rats in the model group fell off;the glands were arranged disorderly;a large number of red blood cells and inflammatory cells gathered,reaching the muscular layer of the mucosa,eroding normal cells;a large amount of red staining in the local area,and a lot of yellow or brown necrosis in the epithelium organization.The gastric mucosa in the traditional Chinese medicine group is arranged regularly,the mucosa is slightly thickened.and there are strips of hemorrhage in the interstitial tissue.In the western medicine group,the gastric mucosal epithelial structure was basically intact;the glands were arranged neatly,accompanied by slight tissue edema;a small amount of red blood cells were diffusely distributed in the interstitial space,and a few vacuole-like changes.The gastric mucosal structure of rats in the AG490 group was basically intact,without obvious gastric mucosal epithelial ulcers or deletions,but there was still a little erosion of the superficial gastric mucosal epithelial tissue,and the epithelial cells were arranged slightly dislocated.However,there is local bleeding,and red blood cell aggregation can be seen between the cells.2.4 JAK2/STAT3 pathway related protein expression:There are little protein content of JAK2 in gastric mucosa,and the signal intensity of p-JAK2 in the model group is slightly higher than that in the blank group(P<0.05),and the phosphorylation of STAT3 is more active than that in the blank group.Compared with the model group,the expression levels of p-JAK2 and p-STAT3 in the Chinese medicine group,the western medicine group and the AG490 group were significantly reduced.Compared with the western medicine group,the expression of p-STAT3 was not significantly different in the traditional Chinese medicine group.ConclusionOne-time exhaustion swimming,incremental training exhaustion swimming and daily exhaustion swimming can all cause exercise-induced stress ulcers in rats.The survival rate and model-building success rate of one-time exhaustion swimming and incremental training swimming are higher than daily Exhausted swimming,and the modeling effect is better.Based on time and economic cost considerations,the one-time exhaustive swimming model is more worthy of recommendation.The occurrence of exercise stress ulcers is related to oxidative stress,which reduces the SOD value in the body and increases the MDA content.Exercise stress ulcer may be caused by activating the JAK2/STAT3 pathway.Inhibiting the phosphorylation of JAK2 and STAT3 can reduce gastric mucosal damage.BXD can inhibit JAK2/STAT3 signaling pathway,reduce the expression of TNF-α,IL-1β,and IL-6 in gastric mucosa,inhibit inflammation,and reduce the occurrence of exercise-induced gastric ulcer. |
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