Influence Of Banxia Xiexin Decoction On Interstitial Cells Of Cajal, Enteric Nervous System And CBS In Diabetic Rats

ObjectiveTo study the effects of Banxia xiexin Decoction treatment on Interstitial Cells ofCajal,Enteric nervous system and CBS of rats with diabetes mellitus.Methods1. Method to obtain diabetic rat models The60healthy male SD rats were dividedinto normal control group, diabetes mellitus model group(DM model group), Banxiaxiexin Decoction group (Chinese herb group) and domperidone group (westernmedicine group).To obtained diabetic rat models, all rats were injected intraperitoneally(ip) streptozotocin (STZ,55mg/kg) except ones in the normal control group. Thesuccessful models were selected in6weeks.2. Intervention methods Daily gavage treatment of Banxia xiexin Decoction [5.4g/(kg·d)] to Chinese herb group, domperidone [5mg/(kg·d)] to western medicine group,and normal saline to the others were lasted in8weeks.3. General situation Eating, drinking, feces, fur color and mental condition of themodel rats were observed periodically. Besides, glucose levels and weight of the ratswere recorded every week.4. Measuring of gastric residual rate and small intestinal propulsive rate Gavagetreatment of semi-solid nutrients (1mL/100g) to the rats obtained in step2that hadbeen fasted by12hours. After30minutes, ligating the pylorus and cardia of the anesthetic rat quickly. Then taking out the stomach, ligating pylorus and ileocecal,isolating mesenteric without tension. The gastric residual rate and small intestinalpropulsive rate were calculated as follows: Gastric residual rate (%)=(Total weight ofthe stomach-Pure weight of the stomach)/(weight of the nutrients)×100％Small intestinal propulsive rate (%)=(Length of the jejunum propulsion)/(Total lengthof the jejunum)×100％The total length of the jejunum was calculated by measuringthe length of the pylorus to the ileocecal. The length of the jejunum propulsion wascalculated by measuring the diffusion length of nutrients.5.Immunohistochemistry Immunohistochemical detection of gastric antrumImmunohistochemical detection of gastric antrum c-kit, SCF and intestinal c-kit, SCF,SYP, PGP9.5, CBS, changes in SP expression were experimented, repectively.6.Expression of c-kit, SCF protein, and CBS protein in jejunum by western blotExpression of c-kit, SCF protein, and CBS protein in jejunum was detected by themethod of western blot. Immunoblotting detection of protein extracted from thejejunum of the rats was experimented. Changes of c-kit and SCF protein expressionwere observed by the gray value.Results1. General situation Weight of the DM model group increased compared to that ofthe normal control group. Besides, both glucose level and gastric residual rate increasd.Symptom of much drinking, eating and urine of the DM group relieved because of themedicine of Chinese herb group and western medicine group. Well mental condition,more weight (P<0.05) and less gastric residual rate (P<0.05) of Chinese herb group andwestern medicine group, lower glucose level (P<0.05) of Chinese herb group wereconfirmed. 2. Immunohistochemical results of antrum, jejunum c-kit and SCF of the diabeticrat The average optical density (P<0.05) of antrum, jejunum c-kit and SCF of theDM model group decreased significantly compared to that of the normal control group;The average optical density (P<0.05) of antrum, jejunum c-kit and SCF of the Chineseherb group and western medicine group increased significantly compared to that of theDM model group. The average optical density (P<0.05) of antrum c-kit of Chinese herbgroup was higher than that of western medicine group.3.Immunohistochemical results of jejunum CBS of the diabetic ratThe optical density (P<0.05) of jejunum CBS of the DM model group increasedcompared to that of the normal control group; The optical density (P<0.05) of s jejunumCBS of the Chinese herb group and western medicine group decreased compared to thatof the DM model group. There was little statistical difference in CBS optical densitybetween Chinese herb group and western medicine group (P＞0.05).4. Immunohistochemical results of jejunum SYP, PGP9.5and SP of the diabeticrat The average area of positive products and absorbance (P<0.05) of SYP, PGP9.5and SP of the DM model group decreased significantly compared to that of the normalcontrol group; The average area of positive products and absorbance (P<0.05) of SYP,PGP9.5and SP of the Chinese herb group and western medicine group increasedsignificantly compared to that of the DM model group. It was lower than that of thenormal control group nevertheless (P<0.05).5. Expression of jejunum CBS, c-kit and SCF protein of the diabetic ratExpression of c-kit protein and SCF protein of the Chinese herb group and westernmedicine group (P<0.05) increased significantly compared to that of the DM modelgroup, It was lower than that of the normal control group nevertheless (P<0.05). There was no obvious difference in expression of c-kit protein and SCF protein betweenChinese herb group and western medicine group (P＞0.05). Expression of CBS proteinof the Chinese herb group and western medicine group was lower than that of the DMmodel group, It was higher than that of the normal control group nevertheless (P<0.05).Conclusions1. There are obstacles in the gastrointestinal motility for diabetic rats.2. Banxia xiexin Decoction contributes to reverse the impaired gastric antrum ICC andgas neurotransmitter H2S according to the fact that it can promote the expression ofc-kit and SCF and decrease the expression of CBS. Because of this, it promotes thegastrointestinal motility obstacles of diabetic rats.3. Banxia xiexin Decoction promotes the expression of jejunum myenteric plexus SYP、PGP9.5and SP. Because of this, it has partial recovery to the impaired jejunummyenteric plexus of the DM rats.4. Abnormal synthesis of H2S may be one reason for the intestinal motility obstaclesaccording to the fact that abnormal expression of jejunum CBS of the diabetic rat wasconfirmed.