| Gastroparesis is in the exclusion of pyloric or duodenal mechanical obstruction cases the emergence of gastric motility disorders and delayed gastric emptying phenomenon,diabetes is the main cause of gastric paralysis.Diabetic gastroparesis(DGP)generally does not endanger the lives of patients,but the occurrence of DGP will interfere with the absorption of nutrients and drug efficacy of patients to the treatment and prognosis of patients with great distress.So the pathogenesis and treatment of DGP increasingly attention.We are based on TCM syndrome differentiation,Clinical application of Chinese medicine of replenishing Qi,nourishing Yin and activating blood and removing stasis treatment of DGP,can improve gastrointestinal motility,reduce the symptoms of patients,and achieved good clinical results.But the mechanism of treatment of diabetic gastroparesis is currently less studied.In this study,we investigated the effect of Chinese medicine of replenishing Qi,nourishing Yin and activating blood and removing stasis on diabetic gastroparesis rats,and observed the effect of migration and activity andutophagy of gastric smooth muscle cell,and to explore the effect and possible treatment of diabetic gastroparesis mechanism.Objective: To observe the effect of Chinese medicine of replenishing Qi,nourishing Yin and activating blood and removing stasis on the autophagy and cell migration of gastric smooth muscle in diabetic gastroparesis,and to explore its role and possible mechanism in the treatment of diabetic gastroparesis.Methods:Thirty male Sprague-Dawley(SD)rats,with body weight of 201-205 grams were randomly divided into three groups: normal control group(NC group),model group(MD group)and Chinese medicine group(ZY group),10 rats in each group.Rats in model group and traditional Chinese medicine group were injected intraperitoneally with STZ to establish diabetic rat model of gastroparesis.After the success of establishing model,the Chinese medicine group to 10 ml /(kg.d)Chinese medicine gavage,once a day.Normal group and model group were given equal volume of saline saline.Continue intragastric administration for 6 weeks.all the rats in the three groups were lavaged with phenol red solution to measure the rate of gastric emptying.Take part of the stomach tissue was frozen in liquid nitrogen and stored in the refrigerator at-80 DEG C.Protein imprinting(Western blot)method to detect Beclin-1?LC3-??JNK and p-JNK protein expression in gastric tissue.We take fresh stomach tissue,in order to extract the primary cells of smooth muscle cells and smooth muscle cell subculture,in order to observe the cell migration ability and cell activity changes.Results:1 Comparison of the general condition of ratsThe body weight of the normal group was increased before the experiment began.The spirit,the eating,the drinking water,the activity,the fur and the urine were normal,and there was no obvious change before the experiment.No rats died during the experiment.However,the other two groups of rats after the injection of STZ,appeared gradually polyphagia,polydipsia,polyuria and the spirit of malaise,activity decreased,the reaction is not sensitive,fur color dim,fecal time is irregular.Comparing with the model group,the traditional Chinese medicine group were improved in the general condition of the spirit,activity,sensitivity and diet.And there were two rats died in the model group,no rats died in the traditional Chinese medicine group.2 Comparison of body weight and blood glucose in each groupComparing with the normal group,the body weight of the model group and the traditional Chinese medicine group were significantly reduced.(P<0.05).Comparing with model group and the traditional Chinese medicine group,there was no significant change in body weight of rats(P>0.05).3 Comparison of rats gastric emptying rateCompared with the normal group,the gastric emptying rate was significantly lower in the model group and the traditional Chinese medicine group(P<0.05).Compared with the model group,the traditional Chinese medicine group was significantly higher in gastric emptying rate(P<0.05)4 Expression of gastric tissue of rats with LC3-??Beclin-1?JNK and p-JNK proteinCompared with normal rats,the expression of Beclin-1 protein in gastric tissue of model group and traditional Chinese medicine group was significantly increased(P < 0.05).Compared with the model group,the expression of Beclin-1 protein in gastric tissue of the raditional Chinese medicine group was significantly decreased(P < 0.05).Compared with normal rats,the expression of LC3-? protein in gastric tissue of model group and traditional Chinese medicine group was significantly increased(P<0.05).Compared with the model group,the expression of LC3-? protein in gastric tissue of the raditional Chinese medicine group was significantly decreased(P < 0.05).Compared with the normal group,the activity of JNK in gastric tissue of model group and traditional Chinese medicine group was significantly enhanced(P < 0.05).Compared with the model group,the expression of JNK protein in gastric tissue of the raditional Chinese medicine group was significantly decreased(P < 0.05).Compared with the normal group,the activity of p-JNKin gastric tissue of model group and traditional Chinese medicine group was significantly enhanced(P < 0.05).Compared with the model group,the expression of p-JNK protein in gastric tissue of the raditional Chinese medicine group was significantly decreased(P < 0.05).5 Comparison of the migration ability of rats gastric smooth muscle cells by Transwell method in each groupCompared with the normal group,the cell migration ability of the model group and the traditional Chinese medicine group was significantly lower than that of the normal group(P < 0.05).Compared with the model group,the migration ability of gastric smooth muscle cells in Chinese traditional medicine group was significantly increased.(P < 0.05).6 Comparison of Cell viability by CCK-8 methodCompared with the normal group,the activity of rats smooth muscle cells in the model group and the traditional Chinese medicine group was significantly lower(P < 0.05).Comparing with model group and the traditional Chinese medicine group,there was no significant change in the activity of gastric smooth muscle cells(P>0.05).Conclusions:1 Chinese medicine of replenishing Qi,nourishing Yin and activating blood and removing stasis can improve the gastric motility and promote gastric emptying in diabetic gastroparesis rats.2 Chinese medicine of replenishing Qi,nourishing Yin and activating blood and removing stasis can improve the gastric smooth muscle cells migration ability of diabetic gastroparesis rats,which may be one of the mechanisms for the treatment of diabetic gastroparesis.3 The mechanism of Chinese medicine of replenishing Qi,nourishing Yin and activating blood and removing stasis can significantly inhibit the autophagy of diabetic gastroparesis gastric smooth muscle,which may be related to the mechanism of the treatment in diabetic gastroparesis. |
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