Carbuncle Of Myogenic Law Treatment Of Peptic Ulcer Experimental And Clinical Studies

Purpose:This topic is for the purpose of seeking the traditional Chinese medicine to disappearing carbuncle and growing muscle method to treat peptic ulcer's curative effect.Through we study and contract in aspects of histopathology, molecular biology,serology and so on,after gastric ulcer rat treatment,as well as in the peptic ulcer patient's curative effect observation,explore to treat the peptic ulcer's functional mechanism and to the clinical application value.Material and method:1 Material1.1 Object of studyAnimal experimentation:SPF SD male rat 70,weight(200±10) g,divided randomly into 7 groups,named the normal control group,the sham-operation group, the model group,kui de kang group,the decoction group,ulcer capsule group, Omeprazole group,each group of 10.Clinical experiment:The gastric mucosa specimen got from the affiliated hospital gastro-endoscope department,the specimen preserved in the liquid nitrogen,include the gastric ulcer 10 examples,the chronic erosive gastritis 10 examples,chronic superficial gastritis 9 examples.Clinical observation:60 outpatients serviced from February 2007 to October 2008 which are conformed to the selected standard,divide into 2 groups randomly. Treatment group are 30 examples,control group are 30.1.2 Medicines and reagentDisappearing carbuncle and growing muscle decoction;Kui de kang granule, Ulcer capsule;Omeprazole tablete;TFF2 upstream and downstream primers; Cimetidine tablete.100%glacial acetic acid,10%solution of chloral hydrate,10%formal dehyde solution,IL-1β,IL-2 ELIASA enzyme immunoassay kit;aprotinin;NO,NOS kit; DEPC;agarose;sniff of bromide(EB);TRIZOL solution;isopropanol;chloroform; DNA-marker DL2000;RT-PCR kit;100%ethanol.1.3 Main experiment instrumentElectric thermostat temperature box,constant temperature oscillator, TDL-5A desktop centrifuge,721B spectrophotometer,Leica RM2135 slicer;Olympus optical microscope;UV-1601 spectrophotometer;INFINITE M200 microplate reader; SN-682 type r Radioimmunoassay Counter;vernier caliper;Milli-Qultrapure water machine;DY89-Ⅰelectric machine glass homogenate;JY92-Ⅱultrasonic cell pulverizer;DU-600 protein,nucleic acid analyzer;BIOFUGE28RS low-temperature high-speed centrifuge;Indeslt low-temperature refrigerator;PE9600 PCR instrument;EPS-300 Bio-Rad;DYCP-33A electrophoresis tank.2 Method2.1 Animal experimentation partImproves the traditional glacial acetic acid burning method,prepares the rat gastric ulcer model.60kg calculated according to the weight of ordinary people,people with 200g of rat body weight conversion ratio of 6.25:1,calculate the corresponding concentration of each drug:Disappearing carbuncle and growing muscle decoction 1.28g/kg,Kui de kang granule 0.2 g/kg,ulcer capsule 0.018 g/kg,Omeprazole tablete 0.2 mg/kg.Each group of experiment rat after making the mold the 2nd day starts to garage,two times every day,altogether 12 days.The normal group,normal raising, does not carry on garage;The sham-operation group,the model group,gavage distilled water 2ml two times every day;The decoction group,garage 1.28g/kg decoction solution 2ml two times every day;Kui de Kang group,garage 0.2g/kg density to Kui de Kang solution 2ml two times every day;The capsule group,garage 0.018g/kg every day the density ulcer capsule solution 2ml two times every day; Omeprazole group,garage 1.28g/kg density Omeprazole solution 2ml in morning, garage distilled water 2ml in everning.Garage for 12 days,after a night of fasting subjects.Rats in each group with 10%chloral hydrate anesthesia,laparotomy,the stomach found in the pylorus, cardia cut at both ends to remove the stomach,gastric juice check;cut along the greater curvature of stomach with a saline wash to clean bench in parietal flattening observed ulcer,and then along the edge of 3mm cut ulcer gastric wall, the production of HE staining of the biopsy.Far from clearing the bowel,omentum find abdominal aorta with a needle into the scalp,blood about 5ml,per ml blood of aprotinin added to 10μl,mixing,cool home two hours;again to 3000 r/m centrifugation for 10 minutes,from the upper levels,into the EP tube,according to group numbers 1-70 mark,frozen reserve.Observation:the general morphology of gastric mucosa,the determination of ulcer area,gastric PH value of the determination,histopathological examination of gastric mucosa,ELISA assay of serum IL-1β,IL-2,nitrate reduction measured NO content was detected NOS levels in serum.2.2 clinical partsGastric mucosa of patients with each of the groups by adding 1ml TRIZOL reagent samples,with shear organization,with cells broken grinder,so the total RNA extraction,reverse transcription reaction,PCR reaction,agarose gel electrophoresis,then receive the reaction of TFF2 drawn strip.The treatment group gives Disappearing carbuncle and growing muscle decoction to treat.Every day one decoction,the water fries,takes juice 300mL, at 3 times the oral.The control group gives the stomach hot clear capsule treatment.Each time 4 grains,daily 3 times.Two groups be treated in for two monthes as one treatment courses,simultaneously take Cimetidine tablete 200mg, three times a day orally.Disable other drugs during treatment,and avoid eating rough,irritating,such as spicy foods,alcohol quit smoking.The main observation include clinical symptoms,changes in fiber endoscopy,relapse rate and adverse reactions.Results:1.Ulcer area of the model group than normal group and sham operation group (P＜0.01),but the group and there are still differences between the normal group (P＜0.01);Capsule Group,Omeprazole Group compared with the model group were significantly different(P＜0.01),the ulcer area of kui de kang group and the decoction group was significantly less than the ulcer area of Omeprazole Group and the capsule groups,the decoction group there are differences in the control group(P＜0.05).Between the acid value of the decoction group and Omeprazole Group are no significant difference(P＞0.05).2.the IL-1βin decoction group was higher than the normal group, sham-operated group,compared with a very significant difference(P＜0.01); decoction group IL-1βcontent was lower than Kui de kang Group,Capsule Group, Omeprazole group,compared with a very significant difference(P＜0.01);the highest content of model group,compared with the other groups there was a significant difference(P＜0.01);Between decoction group and Omeprazole Group are no significant difference(P＞0.05).decoction group IL-2 content lower than the normal group,sham-operated group,compared with a very significant difference(P＜0.01);the IL-2 in decoction group was higher than Kui de kang Group,Capsule Group,Omeprazole Group,compared with a very significant difference(P＜0.01);the minimum content of the model group,compared with the group there was a significant difference(P＜0.01);Between decoction group and Omeprazole Group are no significant difference(P＞0.05).3.The results showed,NO content in decoction group was significantly higher than the of the model group,but also higher than the capsule group,lower than the normal group and sham-operation group;kui de kang group compare with decoction,slightly higher than the latter;NO content of Omeprazole Group higher than the treatment group,and even higher than the normal group and sham-operated group;capsule group is higher than the NO content in the model group only,which is lower than other groups.Decoction of the NOS group were higher than the vitality of the control group,and was significantly higher than the model group;normal group and sham-operation group dynamic highest NOS.4.TFF2 each case are the expression of gastric mucosa,superficial gastritis group to express strong,followed by erosive gastritis group,gastric ulcer group and the weakest expression.Experimental results show that the expression of TFF2 ulcer group than the other two groups,and rotten to the core group have a very significant difference(P＜0.01),with the superficial group also has a very significant difference(P＜0.01);erosive group and superficial group were significantly different(P＜0.05),have statistical significance.5.This observation shows that disappearing carbuncle and growing muscle method treat toxic-heat type peptic have a better efficacy than the control group, low recurrence for six months only one example,in the course of medication did not find heart,liver,kidney and other vital organs and the toxicity of hematopoietic system damage.Conclusion:1.The experimental result may show that disappearing carbuncle and growing muscle decoction can reduce the size of ulcer in rat,promote gastric epithelial cell proliferation,so that the rapid generation of granulation tissue,and promote ulcer healing,and we also believe that disappearing carbuncle and growing muscle decoction also has a ability of inhibit acid.2.Disappearing carbuncle and growing muscle decoction was able to reduce and improve the serum IL-1β,IL-2 level,to ease the release of inflammatory mediators,reduce inflammation,and enhance the defensive function of gastric mucosa in order to promote the healing of ulcers.3.Disappearing carbuncle and growing muscle decoction will effectively enhance the NOS vitality,regulate local blood flow,moderate to increase the generation of NO to near normal levels,regulating gastric acid secretion and maintaining gastric mucosal integrity and defense functions,regulation of gastrointestinal mucosal blood flow,so as to achieve the promotion of ulcer healing.4.TFF2 in gastric ulcer mucosal tissue expressed the lowest level,its expression in gastric mucosal is relate to damage that negative correlation. We can infer its expression should be far lower than normal people.We believe that the TFF2 can protect gastric mucosal barrier to prevent attacks on the role of the invasion factor. 5.We thought that Disappearing carbuncle and growing muscle method is one kind method of curative effect accurate to treat peptic ulcer,and makes certain breakthrough hopefully in the anti-ulcer recurrence aspect.