| Objective: To explore its regulation mechanism on esophageal and gastric motility in rats with mixed reflux esophagitis by observing the effects of clove-depressing qi prescription on serum brain-gut petide,interstitial cells of Cajal(ICCs),stem cell factor(SCF)and tyrosine kinase receptor(c-kit)in lower esophageal tissue of rats.Methods: Thirty-two SD rats were stratified by body weight and randomly divided into two groups: sham operation group(8 rats)and model group(24 rats).The rats in the model of mixed reflux esophagitis received partial duodenum ligation plus cardiomyotomy.One week after the model was established,the remaining surviving rats in the model group were randomly divided into three groups according to body weight stratification: model plus normal saline group,model plus western medicine group,model plus Chinese medicine group.The traditional Chinese medicine group was treated with the granule suspension of clove depressing qi decotion.The western medicine control group was given mosapride citrate dispersible tablets combined with omeprazole capsule suspension.The other groups were given the same amount of normal saline.The course of treatment are coincidently 14 days.After the end of the intervention period,the rats were fasted for 24 hours,but water is allowed.Each rat was given 2% blue dextran for 0.5ml.After 30 minutes,the rats were sacrificed and blood from the abdominal aorta,the stomach contents and the lower esophageal tissues were taken.The pathological changes of esophageal tissues were observed under light microscope.ELISA was used to detect the concentrations of serum GAS,MTL,NO and VIP.The gastric emptying rate of each group was evaluated by measuring the absorbance of residual pigment in the stomach by a microplate reader.The ultra micro-structure of ICCs cells in rat esophageal tissues was observed by electron microscopy.The expression of ckit and SCF protein in esophageal tissue of rats was observed by western blot.The expression of SCF m RNA and c-kit m RNA were observed by real-time PCR.IBM SPSS Statistics 22.0 software was applied and one-way ANOVA was selected for statistical analysis of experimental results.The difference was considered statistically significant if P < 0.05.Results: 1.General state: Before the model was established,there was no significant difference in body weight among the groups.After one week of model establishment,compared with the sham operation group,the body weight of the model group,western medicine group and Chinese medicine group were decreased significantly.Compared with the model group,the body weight of the rats in the western and the Chinese medicine group increased significantly on the seventh and fourth day of intragastric administration(P<0.05,P<0.01).The difference between the Chinese medicine group and the western medicine group was not obvious.2.Pathological changes of rats’ esophageal mucosa: The esophageal mucosa of rats in the sham operation group was normal:no pathological changes were observed except for a small amount of inflammatory cells in the epithelial cell layer.In the model group,squamous epithelial hyperplasia and the mucosal lamina propria extension was obviously visible.A large number of scattered vascular in the submucosa and neutrophils and lymphocytes in the epithelial layer were observed,and a few mucosal erosions and ulcers were seen.Compared with the model group,the inflammatory cell infiltration in the esophageal mucosa of Chinese and western medicine group were significantly reduced,the squamous epithelial hyperplasia and the lamina propria nipple extension were mildly manifested,and a small amount of scattered vasodilation and hyperemia were found.No erosion and ulceration were found.3.Gastric emptying in rats: Compared with the sham operation group,the concentration of pigment in the stomach of the model group increased,and the rate of gastric emptying decreased,the difference was statistically significant(P<0.001).Compared with the model group,the concentration of pigment in the stomach of the rats in the western medicine group and the traditional Chinese medicine group was significantly reduced,and the gastric emptying rate was significantly increased(P<0.01,P<0.05),but the difference between the latter two group was not obvious.4.Changes of serum brain-intestinal peptide concentration in rats: Compared with the sham operation group,the serum concentrations of GAS and MTL in the model group were significantly decreased,and NO and VIP were significantly increased.Compared with the model group,the serum GAS and MTL concentrations in the western medicine group and the Chinese medicine group were significantly increased,with NO and VIP reduced,but the difference between the latter two groups was not significant.5.Ultrastructure of esophageal ICCs in rats: Transmission electron microscopy showed that ICCs in sham-operated rats were mainly distributed in the submucosa and myenteric plexus.ICCs possessed large,elliptical,star-shape or irregular nucleus and less perinuclear cytoplasm,which was rich in cell organelle including endoplasmic reticulum(ER),mitochondria(Mi),Golgi(G),vesicles and other etc.There were typical caveolaes(arrows)on the surface of cell membrane.It was closely connected to the surrounding smooth muscle and nerve fiber ends.The ultrastructure of ICCs in the model group was fuzzy,with abnormal changes such asnuclear pyknosis,enlarged perinuclear space,cytoplasmic lysis,organelle degeneration,mitochondrial swelling,and even vacuolization,endoplasmic reticulum expansion,severe degranulation,and loose gap junctions between ICC and surrounding cells.The above-mentioned performance of the western and Chinese medicine group was not as obvious as the model group.6.Expression of SCF,c-kit protein in rats’ esophagus: Compared with the sham operation group,the expression levels of SCF and c-kit protein in the lower esophageal sphincter of the model group were significantly lower(P<0.0001),and the difference was statistically significant.Compared with the model group,the expression levels of SCF and c-kit protein in the western medicine group were significantly up-regulated in the lower esophageal sphincter(P<0.05),and the SCF and c-kit proteins in the Chinese medicine treatment group were highly expressed(P<0.001,P<0.01).However,the difference between the Chinese medicine group and the western medicine group was not significant(P>0.05).7.Expression of SCF,c-kit gene m RNA in rats’ esophagus: Compared with the sham operation group,the expression level of SCF and c-kit m RNA in the lower esophageal sphincter of the model group was low,and the difference was statistically significant(P<0.001,P<0.0001).Compared with the model group,the expression levels of SCF and c-kit m RNA in the western and Chinese medicine group were significantly up-regulated(P<0.05,P<0.01),but the difference between Chinese and western medicine group was not obvious.Conclusion: 1.Clove descending qi prescription colud increase the GAS,MTL concentration in serum,reduce the content of NO,VIP,and correct neuroendocrine imbalance,thereby promoting gastric emptying,and reinforcing the anti-reflux barrier of the lower esophageal sphincter.2.Clove descending qi prescription may enhancing the pressure of lower esophageal sphincter through up-regulating the transcription and expression levels of SCF,c-kit m RNA and protein in esophageal tissue and promoting the repair of ultra micro-structure of ICCs cells. |
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