| Objective: To study the streptozotocin(STZ) intraperitoneal injection replication of diabetic gastroparesis(diabetic gastroparesis, DGP) model in rats,Given medicine Mazhihuazhuotongluo decoction and cisapride Intervention treatment of the model, using the c-kit antibody as gastric antrum cajal cells(interstitial cell of Cajal, ICC)immunological marker, testing the Motilin and Gastrin content of gastrointestinal hormones in rats by radioimmunoassay, by observing The general conditions of rats, small intestinal impelling in rat, gastric antrum ICC count and MTL, GAS content of hormone expression level, so as to study the effects of Mazhihuazhuotongluo Decoction on diabetic gastroparesis rats small intestinal impelling in rat, gastric antrum distribution of ICC number and MTL, GAS content, to investigate the effect of Mazhihuazhuotongluo decoction on gastrointestinal DGP in rats and the possible mechanism, and to provide the experimental basis for the clinical treatment of DGP by traditional Chinese medicine.Methods: Selected 40 healthy male Wistar rats of clean grade, weight 200±20g, each rat was caged and fed independently, adaptive given ordinary feed for one weeks, the general conditions of rats, were normal for the experiment. The second week 40 rats were randomly divided into normal group(N)8, The other 32 were used to replicate DGP model. Before the experiment the rats stabilized 3 days,fasting 12 h, free drinking water, with 1% of streptozotocin(STZ)(55mg/kg) replicate diabetic rat by a single intraperitoneal injection, after 1 week the diabetic model was successfully replicated by determination of blood glucose concentration higher than 16.7mmol. When the diabetes mellitus model replicated successfully, the 32 rats were randomly divided into four groups, each group 8: model group(M), Mazhihuazhuotongluo decoction group(Z), cisapride group(X), combined Mazhihuazhuotongluo decoction with cisapride group(Z+X). The medicine was given after 1 weeks, Chinese medicine group was given Mazhihuazhuotongluo of traditional Chinese medicine particle prescription 6.3g/kg/d gavage, western medicine group rats were given cisapride 3.6 mg/kg/d by gavage; combined group daily morning dose of medicine given equal by gavage, daily afternoon given the same dose of cisapride by gavage. Normal control group and model group were given an equal volume of saline daily by gavage, once a day. during the experiment the general condition of rats were recorded, the gavage last 6 weeks totaly, After the last administration, the rats were fasted for 24 h, water ban 4h,lavaged the rats with 2ml India blue ink before death, and killed by decapitation, keep the blood specimens form carotid artery in vitro Rapidly. And then immediately took out the whole gastrointestinal tissue in rats and the gastric antrum tissue of rats 0.5cm, detected the small intestinal impelling rate in rat by measured the length from the pylorus to ink front, and from pylorus to ileocecal by a ruler, to observe the pathological changes of intestinal tissue by microscope. Using the c-kit antibody as a specific marker of gastric antrum ICC, the expression and distribution characteristics of c-kit antibody positive ICC was detected by immunohistochemical staining in gastric antrum tissue of rats in each group, The average optical density(A) value of gastric antrum c-kit immune reaction positive products were detected by the FR-980 biological electrophoresis image, the expression levels of the hormone MTL, GAS etc content by radioimmunoassay.Results:1 General states of the ratsNormal rats had good spirit, normal activities, drinking water, food intake, urine volume changed insignificantly. body weight increasing gradually. The model group after modeling appear polydipsia, polyphagia, polyuria, Since then continued, Body weight did not increase, the fur is dryness, edgy yellowing. The general situation of traditional Chinese medicine group, western medicine group, combined group of rats is improved compared with the model group. There was one rat died in the model group due to improper modeling during the entire experiment.2 The changes of body weight in ratsCompared with the normal group, model group, Chinese medicine group, western medicine group and combined group rats’ body weight reduced significantly, the difference was statistically significant(P<0.05); traditional Chinese medicine group, western medicine group, combined group, the body weight of rats increased compared with the model group, the difference was statistically significant(P<0.05); traditional chinese medicine group,western medicine group and the combined group, without significant changes in body weight, no significant difference(P>0.05).3 Changes of blood glucose in ratsThe blood glucose of model group, Chinese medicine group, western medicine group, combined group rats were significantly higher than that in the normal group, the difference was statistically significant(P<0.05); the blood glucose level of Chinese medicine group and combined group decreased than that in the model rats, the difference was statistically significant(P<0.05); The blood glucose of Chinese medicine group and the combined group were decrease, the difference no statistical significance(P>0.05). The western medicine group compared with the model group, the hypoglycemic effect was not statistically significant(P>0.05).4 The hematoxylin eosin(HE) staining of rat gastric antrum smooth muscle tissueThe normal group rats gastric mucosa structure was integrated, and rich in blood vessels, the gland arranged neatly, there were many interstitial cells; the gastric antrum ucosal of Chinese medicine group, western medicine group and combined group were complete, The arrangement was neat, there were abundant blood vessels, there were quantity interstitial cells; The gastric antral mucosa of rats in the model group did not complete, the number of blood vessels was decrease, the glands arranged in disorder, the number of interstitial cells were decreased.5 The small intestinal impelling rates in ratCompared with the normal group, model group, Chinese medicine group, western medicine group and combined group of small intestinal impelling rate of rats decreased, the difference was statistically significant(P<0.05); traditional Chinese medicine group, western medicine group and combined group of small intestine impelling in rats were increased significantly than that in the model group, the difference was statistically significant(P<0.05); traditional Chinese medicine group and the western medicine group of small intestinal impelling rate had no obvious change, no significant difference(P>0.05).6 The ICC expression of rat gastric antrum tissueCompared to normal group, Traditional Chinese medicine group, western medicine group and combined group rats, the distribution of positive cells decreased, the average optical density decreased significantly, the difference was statistically significant difference(P<0.05). the average optical density of the model group reduced and expression decreased(P<0.05), c-kit positive cells can be seen in the deeper color in traditional Chinese medicine group and combined group and Western medicine group, the average optical density value was higher than that of the model group(P<0.05), and the traditional Chinese medicine group and combined treatment group the average light density values were significantly higher than those in the western medicine group(P<0.05); There was no significant difference between Traditional Chinese medicine group and the combined group(P>0.05), but more than in western medicine group(P<0.05).7 The expression level of MTL content of rats in each groupThe traditional Chinese medicine group, western medicine group and combined group the MTL content was obviously lower than that of model group, the difference was statistically significant(P<0.05), there was no significant difference between the three group(P>0.05), there was no significant difference between the combined group and the normal group(P>0.05).8 The expression level of GAS content of rats in each groupCompared with the model group, Chinese medicine group, western medicine group and combined group GAS decreased obviously(P<0.05), in which no obvious differences between the traditional Chinese medicine group and Western medicine group(P>0.05); Combined group compared with the traditional Chinese medicine group and Western medicine group, the content of GAS decreased, the difference was statistically significant(P<0.05), and had no evident statistical significance with normal group(P>0.05).Conclusion:1 Diabetic gastroparesis rats had gastrointestinal motility disorders, Mazhihuazhuotongluo decoction could reduce the level of blood glucose in DGP rats, improve the rat small intestinal propulsion rate, may be this is one of the possible mechanisms for the treatment of DGP.2 Mazhihuazhuotongluo decoction can increase the average optical density rats with DGP of gastric antrum, may be through regulation of the gastric antrum interstitial cells of Cajal(ICC) number, and promote gastric emptying in treatment of DGP rats.3 Treatment of DGP level, Mazhihuazhuotongluo decoction may improve DGP rats MTL, GAS and other gastrointestinal hormones, provides the experimental basis for the treatment of Chinese medicine DGP. |
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