| ObjectiveTo study the effects of Ziyin jiangwei Decoction treatment on Interstitial Cells of Cajal(ICC)、 myenteric plexus of rats with diabetic gastroparesis.To explore some causes of diabetic gastroparesis,and to discuss Ziyin jiangwei Decoction "descending the stomach qi and removing the stomach ivisible lump" and to adjust the mechanism of diabetic gastroparesis.Methods1 Rats with diabetic gastroparesis made models and groupingHealthy male SD rats were divided into experimental group and normal group.The experimental group were made model by intraperitoneal injection with streptozotocin(STZ,60mg/kg) one off.Choosing successful diabetes model,Raising eight weeks,and replicate the rat model with diabetic gastroparesis successfully(Preliminary experiments have confirmed eight weeks in the rat model with diabetic gastroparesis). Choosing stomach yin insufficiency Model rats and randomly divided them into 3 groups:Chinese medicine high dose group, Chinese medicine medium dose group, Chinese medicine low dose group,others were divided into two groups:model group and western medicine group.2 Intervention methodForm 9 weeks all the rats began gavaging.Chinese medicine group were given Ziyin jiangwei Decoction,Low、medium and high dose group, were respectively given concentration 3.125 g(kg·d),6.25 g (kg·d),12.5 g (kg·d) (in accordance with the original medicinal content) of decoction(The low dose group of daily dosage is 1/2dose group, high dose group are 2 times of low dose group).The western medicine group was given 2.92mg/(kg·d) domperidone solution, the model group and normal group were given equal sterile water, all the rats were gavaged once everyday,7 times aweek, a total of 8 weeks.3 General situationObserving all the rats’eating, drinking water, feces, fur colour and mental status periodically.Besides,measuring rat s’tail peripheral blood sugar weekly.4 Measuring small intestine transmission rateand motilinAfter gavaging treatment for 8 weeks, Indian ink 0.5mL were respectively gavaged to rats that had been fasted by 24 hours.After 30 minutes, intraperitonealing injection chloral hydrate (3mL/kg) to anesthesia in rats, opening the abdominal cavity,taking blood 10 ml from abdominal aorta,saving them in -20 ℃ refrigerator.Measuring the length from pylorus to ileocecus as the total length of the small intestine,the length from the pylorus to ink as the length of small intestine propulsion,small intestine transmission rate(%)= length of small intestine propulsion (cm)/total length of the small intestine (cm) x 100%.5 Immunohistochemical staining and IPP image analysisUsing immunohistochemical method to observe the expression of duodenum c-kit, synaptophysin(SYN), Protein gene product 9.5 (PGP9.5). Using IPP 6.0 software for image analysis and data statistics.Results1 General situationCompared with normal group, model groups have the symptom of much drinking, eating and urine, weight reduction and lack of mental state.Compared with model group, the symptom of western medicine group and three Chinese traditional medicine groups reduced relatively, mental state is slightly better.2 Blood glucoseCompared with normal group, before treatment、at the end treatment of 4 weeks and 8 weeks, fasting glucose were significantly higher (P< 0.01).At the end treatment of 4 weeks,compared with model group, treatment group in addition to the high dose group, the rest had no statistical significance (P>0.05);Comparing between the treatment groups, middle and low dose group had no statistical significance (P> 0.05), the two compared with western medicine group had no statistical significance (P> 0.05);Compared with the high dose group, model group 、western medicine group, middle dose group were higher, statistical significance (P< 0.05).At the end treatment of 8 weeks,compared with model group, treatment groups had no statistical significance (P> 0.05).Between the treatment groups, except for the high dose group is lower than western medicine group, the comparison was statistically significant (P< 0.05), other groups had no statistical significance (P>0.05).3 MotilinGroups of rats serum motilin statistical did not see obvious difference (P> 0.05).4 The small intestine transmission rateAt the end treatment of 8 weeks,no significant differences between model group and western medicine group (P>0.05), but were significantly lower than that of traditional Chinese medicine group of high, medium and low three dose groups and normal group,the two groups of model group、the western medicine group had significant difference with the rest of the group (all P< 0.01).Chinese traditional medicine group between high、medium and low three dose groups had no obvious difference (P> 0.05).5 Immunohistochemical staining and image analysis of duodenum c-kit、SYN、PGP9.5Immunohistochemical results showed that each group of duodenum were seen positive cells of c-kit、SYN、PGP9.5, and the positive immune reaction product into a tan.Expression with the highest positive product area and average optical density was normal group (P<0.05);Model group is the lowest (P<0.05).Three chinese medicine groups (TCM) had no obvious difference (P> 0.05).C-kit and SYN had no obvious difference between the group of western medicine and Chinese medicine three dose groups (P> 0.05).PGP9.5 comparison between the treatment group, high dose group is higher than western medicine group, statistically significant (P< 0.05).Conclusion1 Diabetic gastroparesis rat model were copied successfully, and through the small intestine transmission rate determination, verifying the diabetic gastroparesis rats do exist delayed gastric emptying.2 The mechanism of action that Ziyin jiangwei Decoction treats diabetic gastroparesis may not through the motilin, but probably by the way of adjusting the abnormal part of the ICC、 myenteric nerve plexus to improve the symptoms of rats with diabetic gastroparesis,and Ziyin jiangwei Decoction has certain help to drop fall blood sugar.3 The effect of Ziyin jiangwei Decoction was similar to Domperidone,and through comparing different dose groups,which may provide more reference for the TCM therapy and reasonable dosage exploration. |
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