Mechanism Study On Invigorating Spleen And Resolving Dampness Grains Cure Irritable Bowel Syndrome Of Spleen Deficiency

Objective: Invigorating spleen and resolving dampness grains were advanced from clinical decoction, Invigorating spleen and resolving dampness decoction has good curative effect to irritable bowel syndrome of deficiency of the spleen. The aim of this study was to confirm the curative effect of the Invigorating spleen and resolving dampness grains, discuss the mechanism of action of Chinese traditional medicine from it can improve the MTL in plasma,the content of NOS and NO in colon tissue based on the curative effect, systemically open out the effect characteristic of the Invigorating spleen and resolving dampness grains relatively.Methods:1. Invigorating spleen and resolving dampness grains were made by water,low temperature,dynamic extraction; frost condense; spray drying; dry granulation.2.The rats model of irritable bowel syndrome of deficiency of the spleen was made by the mothed of qiaoshi: All the rats are forbidden eating for 10 hours before the experiment, but drinking freely. The model group(M),the positive controll group(PC),the low-dose group(LD),the Mid-dose group(MD) and the High-dose group(HD) of invigorating spleen and resolving dampness grains were given Sennae decoction (0.6g/ml) by intragaster (dose:10ml/Kg), qd. Then binding the back legs of the rats, give them stress and stimulation, last for 2hours.The experiment lasts 2 weeks and the normal group(N) only give water during the experiment.3.The loose stools'grade in 5 hours after given Sennae decoction: the loose stools'grade expresses the grade of the loose stools, distinguished by the smutch's size which was made on the filter paper by the loose stools, it was separate into 8 grades(0,1,2,3,4,5,6,7), stat. the grade of the loose stools one by one, and then got the average grade according to the formula.4.Mothed to measure the advance rate of carbon powder in the small intestines: One and a half hours after the last medicine, give each rat 1ml 5%carbon powder mixed with water, 30minutes later, kill them, get the small intestines, measure the advance rate.5.Mothed to measure the quantity of Motilin(MTL) in blood plasma: Use Radioimmunoassay according to xiejianqun's experiment. Firstly, execute the sample of blood plasma. After give the carbon powder, general anaesthesia by aether, get 2ml blood from the cardinal wein in abdominal cavity, put the blood in the test tube which contents 30μl EDTA-Na2 and 30μl Aprotinin, centrifugate 15minutes with the speed of 3000p/min, take the blood plasma and keep it in -20℃. Secondly, measure the quantity of MTL. All the work must be done according to the instruction of the reagent box.6. Mothed to measure the quantity of Nitric Oxide Synthetade (NOS) and Nitrogen Monoxidum (NO) in colon: using nitrate/nitrite colorimetric assay. Firstly, execute the sample of colon: get a piece of colon from more than 2cm above the anus, weighed, add some isotonic Na chloride to make the tissue bonogenate, centrifugate for 10min with the speed of 3000r/min, keep the supernatant; Secondly, measure the quantity of NOS and NO: measuring by nitrate/nitrite colorimetric assay. All the work must be done according to the instruction of the reagent box.7. Local colon's pathoscopy: take 3 rats from each group randomly, get a piece of colon to make sure no inflammation of pathological change in the mucous membrane of colon.8.Preparation the ex vivo intestinal canal and Taishi liquid: 8.1 Taishi liquid's preparation: get Sodium Chloride (NaCl)8.0g, PotassiumChloride(KCl)0.2g, Sodium Bicarbonate(NaHCO3)1.0g, Magnesium Chloride(MgCl2)0.1g, Sodium Dihydrogen Phosphate(NaH2PO4)0.05g, melt it in 800ml distilled water; Calcium Chloride(CaCl2)0.2g melt in 200ml distilled water, mix it with the formal one slowly, add 1.0g Glucose before used.8.2 Mothed to excute the ex vivo intestinal canal: One and a half hours after the last medicine, give each rat 1ml 5%carbon powder mixed with water, 30minutes later, kill them, get the small intestines, after measuring the advance rate get a piece of intestinal canal about 2cm long, put it in Taishi liquid at time, washed, keep them in -4℃in Taishi liquid.8.3 Turn on the Water bath, Electrophysiolograph and the computer, 0.5h later put the ex vivo intestinal canal on the tensile force transmitter in waterbath with 10ml Taishi liquid, stabled for 0.5h, tracing the cinesi-curve. Measure the frequency and the amplitude. 9.Analysis all the data by One-way ANOVA in the SPSS software, describe it by x±s.Results:1.After 2weeks'therapy, the loose stools'grades and the advance rates of carbon powder in the small intestines of invigorating spleen and resolving dampness grains group and invigorating spleen and resolving dampness soup group are significantly decreased compared with the model group of irritable bowel syndrome of deficiency of the spleen, the curative effects are better than the masculine controlled group. But there is no differences between the two groups.2. After 2weeks'therapy, the loose stools'grades and the advance rates of carbon powder in the small intestines of invigorating spleen and resolving dampness grains'MD and HD groups are significantly decreased compared with the M group of irritable bowel syndrome of deficiency of the spleen. The PC group and the LD group'loose stools'grade and the advance rate of carbon powder in the small intestines are decreased too, but no better than the MD and HD group.3.The quantity of MTL in the blood plasma is significantly decreased compared with the M group after 2week'therapy by Mid-dose and High-dose invigorating spleen and resolving dampness grains, it is decreased too compared with the PC group and the LD group, the difference is significant. The invigorating spleen and resolving dampness grains can improve the level of MTL in blood plasma of irritable bowel syndrome of deficiency of the spleen rats. 4.The level of NOS and NO in local colon tissue of Mid-dose,High-dosegroups is significantly higher than the M group and the PC group, the difference is significant. The invigorating spleen and resolving dampness grains can improve the level of NOS and NO in local colon tissue of irritable bowel syndrome of deficiency of the spleen rats.5.The amplitude of ex vivo intestinal canal in irritable bowel syndrome of deficiency of the spleen rats is significantly decreased compared with the M group after 2 weeks'therapy by Mid-dose,High-dose invigorating spleen and resolving dampness grains. The amplitude in the PC group is decreased too compared with model group, but no differences in statistics. There are some quantity-effect relationships between the 3 invigorating spleen and resolving dampness grains groups.Conclusions:1. Invigorating spleen and resolving dampness grains and invigorating spleen and resolving dampness soup all have good therapy effect to irritable bowel syndrome of deficiency of the spleen.2. Invigorating spleen and resolving dampness grains has good therapy effect to irritable bowel syndrome of deficiency of the spleen. The mechanism is to achieve the therapy aim by improving the level of MTL in blood plasma and the level of NOS,NO in local colon tissue.