Experimental Research About Substance Basis And Mechanism Of Qi-regulating Class Prescriptions In Treating Functional Dyspepsia(FD)

Objective: The main objective of this research is: 1 To attempt to make FD animal model with"irregular food intake match with clamping the tail and stimulation method". 2 To research the Substance Basis and Mechanism of Qi-regulating Class Prescriptions(Sinisan, Zhizhuwan, Xiaoxianxiongtang) in treating FD, discuss the scientific evidence of"regulating activities of qi"method in treating FD.MethodsExperment 1: The improvement of the FD animal modelThe 1st step: To make the model32 health apinoid male Wistar rats, mean weight of 200g, were divided into 4 groups(8 of one group), named: 1.Normal group(NG): Fed by normal ways. 2. Irregular food intake group(IG): Fed in odd days not even days. 3. Clamped the tail and stimulation group(CG): Fed by normal ways and clamped the 1/3 extremity of the tail with long forceps, each time lasted 30 minutes, between two times was half an hour's interval; whole jobs ended after the 4th hour. 4. Irregular food intake match with clamping the tail and stimulation group(ICG): Fed in odd days and clamped the tail in even days, each time lasted 10 minutes, between two times was an hour's interval; whole jobs ended after the 4th hour. Observed the change of the food and drink amount, weight, color of capill, dejecta and temper.The 2nd step: To verificate the modelLeft the rats durining 24 hours without feeding and drinking. Then fill them with amylum pasta(1g/ml) by intragastric administration (10ml/kg). After 1 hour's standing, killed them by decapitation. Cut the belly open, deligated the cardia and janitor, took out of the stomach and weighted it. Then cut the stomach open, washouted the content by distilled water, blot it by filter paper, then weighted the empty stomach. Calculated the empty rate, and observed the stomach mucosa.Experment 2: Experimental research about substance basis and mechanism of Qi-regulating class prescriptions in treating FDThe 1st step: To make the model and therapy48 health apinoid male Wistar rats, mean weight of 200g, were divided randomly into 6 groups(8 of one group). Selected 1 group as blank contrast group(BG), fed by normal ways for 14 days. Other 5 groups were made models by"irregular food intake match with clamping the tail and stimulation method"for 14 days. Then filled the BG rats with normal saline by intragastric administration(10ml/kg) for 14 days; and named other 5 groups as: model contrast group(MG), Sinisan group (SG), Zhizhuwan group(ZG), Xiaoxianxiongtang group(XG) and Domperidone group(DG), filled respectively with normal saline, Sinisan decoction, Zhizhuwan decoction, Xiaoxianxiongtang decoction and Domperidone liquor by intragastric administration for 14 days. Observed the change of the general condition and the amount of food and drink.The 2nd step: To detect the indicatrixesExperment of stomach emptying: Left the 6 groups durining 24 hours without feeding and drinking. Then fill them with amylum pasta(1g/ml) by intragastric administration (10ml/kg). After 1 hour's standing, killed them by decapitation, gathered 4ml of blood, put into the anticoagulation tube and placed in the refrigerator, waited for detecting NO, AchE and MTL. Then cut the belly open, took out of the stomach and weighted it. Then cut the stomach open, washouted the content, weighted the empty stomach. Calculated the empty rate, and observed the stomach mucosa. Cut the sinus ventriculi and put it into formaldehyde liquor of 10% for fixation, the remanent stomach tissue were reserved and levigated into homogenate of 10% waited for detecting No and AchE.The NO of the stomach tissue and blood serum had been detected by"nitrate reductase method"; the AchE of the stomach tissue and blood plasma had been detected by"colorimetric method"; the MTL of the blood plasma had been detected by"radioimmunoassay method".To observe the constitution of the sinus ventriculi tissue: Take out the sinus ventriculi(reserved at the experment of stomach emptying) from the formaldehyde liquor, dewatered by gradient alcohol, made clearing by dimethyl benzene, embeded by ozocerite, made microtome section in 4μm's thickness, stained by hematoxylin and eosin, observed the constitution of the sinus ventriculi tissue by the light microscope.ResultsExperment 11 The general condition and stomach mucosa The NG rats had steady amount of food and drink, pure white and soft capill; tectonic dejecta without thick smell; gentle temper and good mental status, erythroic colored and unabroken stomach mucosa. There were no significant change in the IG rats about the items above-mentioned. CG and ICG rats had indiscriminate amount of food and drink, yellow and dry capill; deformed and foul dejecta; crabby temper and bad mental status. The stomach mucosa of them were congested and edematus. Ulcer and erosion could be seen in CG rats'stomach mucosa.Compared about the increasing quantities of the weight (g): IG(22.99±4.51), CG(12.26±5.32) and ICG(17.54±8.77) were all significant lower than NG(33.71±7.45) (P<0.01). Compared among the model groups, CG was significant lower than IG(P<0.01).2 Compared about the stomach empty rate (%)Compared with NG(27.00±4.06), CG(15.94±4.09) and ICG(20.88±1.19) were both significant lower (PCG<0.01,PICG<0.05). Compared among the model groups, CG was significant lower than IG(24.75±1.10) (P<0.01).Experment 21 The general condition and stomach mucosaBG rats had steady amount of food and drink; significant increasing of the weight; pure white and soft capill; tectonic dejecta without thick smell; gentle temper and good mental status; erythroic colored and unabroken stomach mucosa. MG rats had indiscriminate amount of food and drink; yellow and dry capill; deformed and foul dejecta; crabby temper and bad mental status; congested and edematus stomach mucosa without impairment. DG and three Chinese herbal groups were as same as MG before treatment; while after treatment, improvement had taken place about the amount of food and drink, weight, capill color and dejecta; with the gentler temper, better mental status and erythroic colored and unabroken stomach mucosa.2 Compared about the stomach empty rate (%)Compared with MG(16.25±3.82), Sinisan(26.19±0.84),Zhizhuwan (25.50±1.00) and Xiaoxianxiongtang(25.38±1.19) all could significant increase the stomach empty rate(PSG<0.01,PZG, XG<0.05); Sinisan was significant better than Domperidone(22.69±0.96) (P<0.01). The three Chinese herbal prescriptions were equipotent (P>0.05).3 Compared about the NO of stomach tissue and blood serumStomach tissue (μmol/gprot): Compared with MG(4.08±0.23), Sinisan(2.80±0.20), Zhizhuwan(3.10±0.24) and Xiaoxianxiongtang (3.39±0.14) all could significant decrease the NO (P<0.01), and were all significant better than Domperidone(3.68±0.23) (P<0.01)。Compared among the three Chinese herbal prescriptions, the effectiveness was (from high to low): Sinisan, Zhizhuwan, Xiaoxianxiongtang (P<0.05).Blood serum (μmol/L): Compared with MG(59.75±10.96), Sinisan (42.56±4.55), Zhizhuwan(45.53±5.34) and Xiaoxianxiongtang(46.64±10.37) all could significant decrease the NO (P<0.05), and were equipotent with Domperidone(47.52±9.59) (P>0.05). The three Chinese herbal prescriptions were equipotent (P>0.05).4 Compared about the AchE of stomach tissue and blood plasmaStomach tissue (U/mgprot): Compared with MG(0.74±0.11), Sinisan(0.88±0.07), Zhizhuwan (0.87±0.06) and Xiaoxianxiongtang (0.84±0.06) all could significant increase the AchE activity (P<0.05), therefore increase the Ach propotional. And were equipotent with Domperidone(0.85±0.09) (P>0.05). The three Chinese herbal prescriptions were equipotent (P>0.05).Blood plasma (U/ml): Compared with MG(7.02±0.90), Sinisan (11.43±1.80), Zhizhuwan(9.96±2.20) and Xiaoxianxiongtang (8.96±1.60) all could significant increase the AchE activity (PXG<0.05, PSG, ZG<0.01), therefore increase the Ach propotional. Sinisan was significantly better than Domperidone(9.37±2.33) (P<0.05). Compared among the three Chinese herbal prescriptions, Sinisan was significantly better than Xiaoxianxiongtang (P<0.01).5 Compared about the MTL of the blood plasma (pg/ml)Compared with MG(385.21±10.98), Sinisan(430.04±8.25), Zhizhuwan(422.04±12.46) and Xiaoxianxiongtang (417.67±9.05) all could significant increase the MTL (P<0.01). Sinisan and Zhizhuwan were significant better than Domperidone(408.29±11.11) (PSG<0.01,PZG>0.05). Compared among the three Chinese herbal prescriptions, Sinisan was significant better than Xiaoxianxiongtang (P<0.05).6 The constitution of the sinus ventriculi tissueMG rats'mucosas of sinus ventriculi were unabroken, with congestion, edema and infiltrating of inflammatory cells. DG and three Chinese herbal groups'mucosas were unabroken, without such appearance above-mentioned.Conclusion1"Irregular food intake match with clamping the tail and stimulation method"is better than other departed methods in making FD animal model. This method is similar as the character of FD; the manipulation is convenient and the cycle time is not very long. Significant changes have taken place in the general condition, stomach mucosa and empty rate. It is easy to control and avoid unexpected condition.2 Sinisan, Zhizhuwan and Xiaoxianxiongtang can improve FD rat's general condition and stomach mucosa appearance; increase it's stomach empty rate; lessen the NO's inhibition of stomach empty; increase the Ach and MTL to promote the stomach empty. Sinisan's integrity effect is the best, significantly better than Domperidone in decreasing the NO of stomach tissue and increasing the AchE and MTL of blood plasma. Zhizhuwan is better than Domperidone in decreasing the NO(stomach tissue) and increasing the MTL (blood plasma). Xiaoxianxiongtang is equipotent with Domperidone. All of them can be adopted by differentiation of symptoms and signs to enhance the therapeutic effect in clinically work.