| Objectiveto establish a rat model of chronic physical and psychological stressstress, observe the effect of jiaweisinisan(JWSNS)on behavior, the rateof gastric emptying, intestinal propulsion ratio, electrogastrography,gastrointestinal neurotransmitter, and explore different aspectsmechanism of JWSNS interventioning stress-induced gastrointestinaldysfunction. At the same time, set up chronic physical and psychologicalstress rat model, and observe the effect of JWSNS on gastric mucosa, gastricmucosal EGF, EGFR protein expression, gastric mucosa trefoil factor1(TFF1), endothelin1(ET-1) mRNA expression of chronic physical andpsychological stress rat model, from the morphological structure to observethe effect of JWSNS on chronic physical and psychological stress rats, anddiscuss the mechanism.MethodsThe first part:60male Wistar rats were randomly divided into6groups,namely the normal group, model group, cisapride group, JWSNS high,middle,low doses groups,10rats in each group.Except the blank group, the otherfive groups were given physical and mental stress to make model, molding4week continuously. From third weeks on, at the same time,supplementedJWSNS high, middle,low dose group with JWSNS to gastric lavage and treat,cisapride Group were supplemented cisapride suspension to gastric lavage and treat; blank, model group were treated with normal saline to gastriclavage and treat.(1) During molding, rats were observed and recorded dailyincluding mental state, excitement level, emotional response, behavioralstate, active status, sleep behavior, skin and hair color and condition,everyday observation and record the water intake and food intake amountof every rat respectively, in the day before the experiment and in the7th,14th,21th,28th d after test,use electronic balance to weigh each ratbody weight, body mass changes observed in rats; when molding end, byopen-field test (Open-field) determine cross grid movement times andupright exercise times of experimental rat in the lab and in swimming poolmeasure swimming exhaustive time of experimental rat.(2) After the JWSNSintervention,in the4th weekend, detection gastrointestinal motility inrats, use biological and functional experimental system to observationelectrogastrogram of test rats, and understand the gastric electricalactivity in amplitude and the difference in frequency of the normal group,model group, JWSNS high,middle, low dose group, cisapride group rats. aftertesting, the gastrointestinal motility and electrogastrogram data in thenormal group, model group, JWSNS high,middle, low dose group and cisapridegroup were given corresponding statistical analysis respectively.(3)After stress model,immunohistochemical determination was used todetermine gastric mucosa COX-2, colonic mucosal c-kit average values ofoptical density changes of experimental rat.The second part:60male Wistar rats were randomly divided into6groups,namely the normal group, model group, the omeprazole group, JWSNS high,middle,low dose groups,10rats in each group.Except the blank group, theother five groups were given physical and mental stress model, continuousmolding for4weeks. From third week on, during making model, supplementedJWSNS high, middle,low dose groups with JWSNS to gastric lavage and treat, supplemented omeprazole suspension to omeprazole group to gastric lavagetreatment; blank and model group were lavaged and treated with normal saline.(1) After stress molding end,gastric mucosal morphology and structure,gastric acid and gastric mucosal blood flow were detectioned.(2) Afterstress molding end,gastric mucosal EGF and EGFR protein expression weredetectioned.(3) After stress model,RT-PCR method was used to determinationthe gastric mucosa trefoil factor1(TFF1) and endothelin1(ET-1) mRNAexpression in rat model.ResultsThe first part experiment results:(1) All the rats before the experiment showed a good mental state,and with the stress time tend, third week later, the amount of drinkingwater and food intake in model group rats showed a significant downwardtrend, but after JWSNS each dose group and cisapride group were treated,the decreased tendency of drinking water and food intake were slower indifferent degree.Later period to fourth weeks, compared with the modelgroup, after drug intervention in third and fourth weeks, the drinking waterand food intake of JWSNS each dose group and cisapride group increased indifferent degrees, and there was significant difference (P＜0.05or P＜0.01); and compared with the cisapride group, drinking water and foodintake in JWSNS high dose group rats increased significantly, and therewas significant difference (P＜0.05); after the fourth weekend of themodel, compared with the model group, the weight growth in JWSNS each dosegroup and cisapride group increased to varying degrees, difference thereis significant (P＜0.05or P＜0.01); and compared with the cisapridegroup, in fourth weekend, the body weight of JWSNS high dose group increasedsignificantly, and there was significant difference (P＜0.05); in fourthweekend, compared with the model group, the cross grid movement and vertical movement scores in JWSNS each dose group and cisapride group weresignificantly increased, and there was significant difference (P＜0.05or P＜0.01); and compared with the cisapride group, the cross gridmovement and vertical movement scores of JWSNS high dose group weresignificantly increased, and there was significant difference (P＜0.05or P＜0.01); after fourth weekend of model, compared with the model group,exhaustive swimming time of JWSNS high, middle, low dose groups andcisapride group extended in different degree, and there was significantdifference (P＜0.05or P＜0.01), and compared with the cisapride group,exhaustive swimming time of JWSNS high dose group were significantlyincreased, and there was significant difference (P＜0.05).(2) compared with the model group, the rate of gastric emptying andsmall intestinal propulsion ratio of JWSNS each dose group and cisapridegroup were significantly increased, and there was significant difference(P＜0.05or P＜0.01); and compared with the cisapride group, rate ofgastric emptying of JWSNS high dose group was not elevated (P＞0.05),and small intestinal propulsion ratio increased significantly (P＜0.05);compared with the model group, slow wave rhythm, frequency, power, wavefrequency, gastric slow wave amplitude of JWSNS each dose group andcisapride group increased, there was a significant difference (P＜0.05,P＜0.01), wherein gastric waves of JWSNS high dose group were near normallevels; and compared with the cisapride group, slow wave rhythm and fastfrequency of JWSNS high dose group rats were significantly enhanced (P＜0.05or P＜0.01);(3) compared with the model group, mean optical density of gastricmucosa COX-2of the normal group, JWSNS each dose group and cisapride grouprats decreased, and there was significant difference (P＜0.05or P＜0.01); and compared with the cisapride group, mean optical density of gastric mucosa tissue COX-2in JWSNS middle,high dose groups rats decreased,and there was significant difference (P＜0.05or P＜0.0l). Comparedwith the model group, colonic mucosa c-kit average optical density valuesof the normal group, JWSNS each dose group and cisapride group ratsincreased in different degree, and the difference was significant (P＜0.05or P＜0.01); and compared with the cisapride group, colonic mucosatissue c-kit average optical density values of JWSNS middle,high dosegroups rats increased, and there was a significant difference (P＜0.05or P＜0.0l).The second part experiment results:(1) under the effect of JWSNS, compare the change of gastric mucosalblood flow: compared with the model group, the gastric mucosal blood flowin JWSNS each dose group and omeprazole group rats increased in differentdegree, and the difference was significant (P＜0.05or P＜0.01); andcompared with the omeprazole group, gastric mucosal blood flow of JWSNSmiddle,high dose groups rats increased, and there was a significantdifference (P＜0.05or P＜0.0l). under the effect of JWSNS, comparethe change of gastric juice acidity (pH value) change comparison: comparedwith the model group, gastric juice pH of JWSNS each dose group andomeprazole group rats increased in different degree, and the differencewas significant (P＜0.05or P＜0.01); and compared with the omeprazolegroup, gastric juice pH value of JWSNS middle,high dose groups rats wasnot elevated,and without significant difference (P＞0.05). under theeffect of JWSNS, compare the change of of gastric mucosa action in ratsunder the naked eye: gastric mucosa damage loss in JWSNS high dose grouprat is most obvious, and was close to the normal level, gastric mucosa ofthe omeprazole group, JWSNS middle and low dose group rats were in differentdegree improved. under the effect of JWSNS, compare the change of gastric mucosa ulcer index (UI): compared with the model group, gastric mucosaulcer index (UI) of JWSNS each dose group and omeprazole group ratsincreased in different degree, and the difference was significant (P＜0.05or P＜0.01); and compared with the omeprazole group, gastric mucosaulcer index (UI) of JWSNS high dose groups rats increased significantly,and there was significant difference (P＜0.05). under the action ofJWSNS, the comparison of gastric mucosa pathological observation andinjury severity rating change of each group rat: gastric mucosal bleedingnecrotic ulcer is apparent in the rats of the model group. microscopicallygastric mucosal lesion degree of JWSNS high dose groups rats,compared withthe model group,was significantly reduced, gastric mucosaes in JWSNSmiddle and low dose group were in different degree improves; under theeffect of JWSNS, compare the change of gastric mucosal epithelial cellsultrastructure: in the model group rats, the gastric epithelial cellnecrosis was evident. gastric mucosal epithelial cells of JWSNS high dosegroups rats were closely connected, the structure of cells membrane areintegrity, and cell morphology were in normal. In Omeprazole group rat,thegastric mucosa epithelial cell membrane structure were integrity basicly,cytoplasmic organelles were in the lack of uniform distribution. In JWSNSmiddle and low dose group rats,gastric mucosa epithelial cells return tonormal in different levels；(2) the comparison of gastric mucosa tissue EGF and EGFR proteinaverage optical density value change of all experiment groups: comparedwith the model group, gastric mucosa tissue EGF protein average opticaldensity of JWSNS each dose group and omeprazole group rats decreased, andthere was significant difference (P＜0.05or P＜0.01); and compared withthe omeprazole group, gastric mucosa EGF protein average optical densityvalue of JWSNS middle,high dose groups rats decreased, and there was significant difference (P＜0.05or P＜0.0l); compared with the modelgroup, gastric mucosa average optical density values of EGFR protein inJWSNS each dose group and omeprazole group rats decreased, and there wassignificant difference (P＜0.05or P＜0.01); and compared with theomeprazole group, gastric mucosa EGFR protein average optical density valueof JWSNS middle,high dose groups rats decreased, and the difference wereof significant meaning (P＜0.05or P＜0.0l)；(3) compared with the model group, gastric mucosa trefoil factor1(TFF1) relative expression levels in JWSNS each dose group and omeprazolegroup rats increased in different degree, and there were significantdifferences (P＜0.05or P＜0.01); and compared with the omeprazole group,gastric mucosa trefoil factor1(TFF1) the relative expression levelsof JWSNS middle,high dose groups rats increased in different degree, andthere were significant differences (P＜0.05or P＜0.0l);Compared withthe model group, gastric mucosa endothelin1(ET-1) mRNA relativeexpression levels of JWSNS each dose group and omeprazole group ratsdecreased, and there was significant difference (P＜0.05or P＜0.01);and compared with the omeprazole group, gastric mucosa endothelin1(ET-1)mRNA relative expression levels of JWSNS middle,high dose groups ratsdecreased, and there was significant difference (P＜0.05or P＜0.0l)；ConclusionFrom the first part,we can know that:(1) JWSNS can improve significantly behavioral abnormalities ofchronic stress rat model, and lift depression of model rat caused by stress；(2) JWSNS can significantly improve gastric wave anomaly of chronicpsychological stress rats, promote gastric emptying and small intestinalpropulsion.Its role may be realized through promoting gastric electricalactivity, increasing gastric smooth muscle contraction and other pathways, at the same time,JWSNS promote gastrointestinal motility of rats withdose-effect effects relationship.(3) JWSNS can improve significantly gastrointestinal neurotransmitterabnormalities of chronic stress model rats, ease gastrointestinaldysfunction of a rat model resulted in due to gastrointestinalneurotransmitter disorders.From second part, we can know that:(1) JWSNS can improve significantly gastric mucosal morphology andstructure, gastric acid and gastric mucosal blood flow abnormalities ofchronic stress rat model, and lift gastric mucosal injury of the rat modelleaded to due to chronic stress.(2) JWSNS can improve significantly gastric mucosal tissue cellfunction of chronic stress rat model, and lift gastric mucosa tissue EGF,EGFR protein abnormal expression caused by stress in a rat model.(3) JWSNS can improve significantly abnormal expression of gastricmucosa trefoil factor1(TFF1), endothelin1(ET-1) mRNA of chronicstress rat model, and lift gastric mucosal damage caused by stress in arat model. |
PDF Full Text Request