| Objective:By screening the best concentration of glacial acetic acid to establish rat models of gastric ulcer with cold syndrome; and observing the effect of five alpinia herbs on the behavioral signs, gastric juice, gastric acid, proteases, and gastric histopathological changes; to explain the function of warming stomach and dispelling cold of these five alpinia herbs; by measuring the concent of PGE2, EGF, TGF-α in stomach tissue, the concent of NO, IL-8, TNF-α, NT, ET in blood serum, the concent of AchE,5-HT, NE in brain, to explore the mechanism of these five alpinia herbs on the treatment of gastric ulcer with cold syndrome.Methods:Rats were divided into five groups, and seperately given glacial acetic acid in the concentration of 5%,10%,15%,20%,25% and cold rhizoma anemarrhenae decoction, to establish rat model of gastric ulcer with cold syndrome, ulcer area, ulcer index, dead number, pathological changes and the natural repair were observed and compared, to determine the optimal concentration of glacial acetic. After determining the optimal concentration of glacial acetic acid, 140 rats were divided into 14 groups, including bland control group, model control group,2 positive control groups,10 herbs groups,10 rats per group. Except blank control group, the other groups were induce by the glacial acetic acid with optimal concentration and Rhizoma Anemarrhenae decoction of 4℃, then the blank control group and model control group were fed with distilled water, the other groups were were fed with test herbs. Behavior symptoms were observed, such as food intake, weight, hair color, stool, and mental status; and the amount of gastric juice, gastric acid, pepsin vitality were measured; pathological changes of the stomach were observed by HE staining; the concent of PGE2, EGF, TGF-α, IL-8, TNF-β, NT, ET,5-HT, and NE were tested by enzyme-linked immunosorbent assay, the NO content were tested by the method of nitrate reduction, the concent of AchE were tested by the method of protein quantification. SPSS 19.0 software was used for statistical analysis, P<0.05 was considered to be statistically significant.Results:1. The establishment of gastric ulcer rat model with cold syndrome by cold herbs and glacial acetic acid:15% glacial acetic acid had the best effect, including making large area of ulcers, no lethal, slow and natural recovery. The best extablishment method was following:Rats were fed with rhizoma anemarrhenae decoction of 4℃ and 15% glacial acetic acid,1 times/d, the rats were fed with rhizoma anemarrhenae decoction at 8:00, and fed with glacial acetic acid at 18:00, for 4d. Indoor environment was set at 16℃ and 60% humidity.2. Studies on the efficacy of five alpinia herbs on the treatment of rat models of gastric ulcer with cold syndrome(1) Effect on the behavior symptoms① Effect on body weight:compared with model control group, the body weight of 10 herbs groups increased (P<0.05 or P<0.01).② Effect on the food-intake:compared with model control group, the food-intake of 10 herbs groups increased, the rate of increase was obvious.③ Effect on the coat color scores:compared with model control group, except high dosage of rhizoma galangae group, the coat color scores of the other groups decreased (P<0.05 or P<0.01).④ Effect on feces and mental state:The feces of tested herbs groups decreased and were dry, took shape, the mental state recovered in some extent, action was more flexible.(2) Effect on the amount of gastric juice, gastric acid, pepsin activity: compared with the model control group, high dosage galanga, high dosage galangal, low dosage galangal, high dose grass cardamom, low dosage grass cardamom, high dosage fructus galangae, low dosage fructus galangae, and high dosage alpinia oxyphylla could reduce the the amount of gastric juice (P<0.05 or P<0.01); high dosage galanga, high dosage galangal, low dosage galangal, high dose alpinia, low dosage grass cardamom, high dosage fructus galangae, low dosage fructus galangae, high dosage alpinia oxyphylla, and low dosage alpinia oxyphylla could increase the pH of gastric acid, and reduce the activity of pepsase (P<0.05 or P<0.01).(3) Effect on gastric tissue pathological changes:compared with the model control group; in high dose galanga group, inflammatory cell infiltration, mucosal surface exudation reduced, granulation tissue increased, New epithelial cells in mucosal surface was arranged; in high dosage galangal group, the necrotic tissue of the surface reduced,muscular submucosal mucous membrane structure was clear, granulation tissue increased, mucosal epithelial tissue lined up; in high dose grass cardamom group, inflammatory cells infiltration decreased, surface exudate decrease, granulation tissue increased, surface epithelial repair began to appear; in high dosage fructus galangae group, inflammatory cells infiltration reduced, granulation tissue were distributed evenly, mucosal exudation of disappeared, new epithelial tissue gradually covered the mucosal surface; in high dosage alpinia oxyphylla group, inflammatory cell infiltration decreased, epithelial began repair in part of the mucosal surface; in high dosage galanga group, mucosal exudate decreased, granulation tissue increased, mucous membrane surf ace was covered in new glands; in low dosage galanga group, mucosal exudate reduced, mucosa epithelial tissue covered the newborn; in low dosage grass cardamom group, muscular layer, the structure of the submucosa was clear, fracture of mucosal muscularis began to repair, mucosal epithelium tissue; in low dosage fructus galangae group, mucosal exudate reduced, inflammatory cells infiltration reduced, granulation tissue increased, mucosal muscularis began to repair; in low dosage alpinia oxyphylla group, the structure of muscular layer was integrity, submucosa was full of granulation tissue.3. Study on the mechanism of the function of warming stomach for dispelling cold of five alpinia Herbs on gastric ulcer rat model with cold syndrome(1) Effect on the PGE2 content in gastric tissueCompared with the model control group, the PGE2 content of high dosage galangal group and high dosage grass cardamom group signifcantly increased (P<0.01), the PGE2 content of high dosage galanga group, low dosage grass cardamom group and low dosage fructus galangae group signifcantly increased (P<0.05), low dosage galanga group, low dosage galangal group, high dosage alpinia oxyphylla group, and low dosage alpinia oxyphylla group had no significant changes (P>0.05).(2) Effect on the content of EGF in gastric tissueCompared with the model control group, the EGF content of gastric tissue of high dosage galanga group, high dosage grass cardamom group, high dosage alpinia oxyphylla group increased signifcantly (P<0.01); the EGF content of gastric tissue of high dosage galangal group, low dosage grass cardamom group, high dosage fructus galangae group increased signifcantly (P<0.05); the EGF content of gastric tissue of the EGF content of gastric tissue of, low dosage fructus galangae group, and low dosage alpinia oxyphylla group had no obviouly change (P>0.05).(3) Effect on the content of TGF-a in gastric tissueCompared with the model control group, the content of TGF-α of high dosage galanga group, low dosage galanga group, high dosage galangal group, high dosage grass cardamom group, high dosage fructus galangae group, and high dosage alpinia oxyphylla group increased signifcantly (P<0.01); the content of TGF-a of low dosage galangal group, low dosage grass cardamom group, low dosage fructus galangae group, and low dosage alpinia oxyphylla group had no obviou change (P>0.05).(4) Effect on the levels of NO in serumCompared with the model control group, the content of NO of high dosage fructus galangae group, fuzilizhong pills group and cimetidine group increased signifcantly (P<0.01); the content of NO of high dosage galanga group, high dosage galangal group, high dosage grass cardamom group, low dosage fructus galangae group, high dosage alpinia oxyphylla group increased signifcantly (P<0.05); the content of NO of low dosage galanga group, low dosage galangal group, low dosage grass cardamom group, and low dosage alpinia oxyphylla group had no significant change (P>0.05).(5) Effect on the content of IL-8 in serumCompared with the model control group, the content of IL-8 of high dosage grass cardamom group, high dosage fructus galangae group, high dosage alpinia oxyphylla group, fuzilizhong pills group, and cimetidine group decreased signifcantly (P<0.01); the content of IL-8 of high dosage galanga group, low dosage galanga group, low dosage galangal group, low dosage grass cardamom group, low dosage fructus galangae group, low dosage alpinia oxyphylla group increased signifcantly (P<0.05)(6) Effect on the content of TNF-a in serumCompared with the model control group, the content of TNF-a in serum of high dosage galanga group, low dosage galanga group, high dosage galangal group, low dosage galangal group, high dosage grass cardamom group, low dosage grass cardamom group, high dosage fructus galangae group, low dosage fructus galangae group, high dosage alpinia oxyphylla group, and low dosage alpinia oxyphylla group had no significantly change (P>0.05).(7) Effect on the content of NT in serumCompared with the model control group, the content of NT in serum of high dosage galanga group, low dosage galanga group, high dosage galangal group, low dosage galangal group, high dosage grass cardamom group, low dosage grass cardamom group, high dosage fructus galangae group, low dosage fructus galangae group, and high dosage alpinia oxyphylla group increased significantly (P<. 0.01); the content of NT in serum of low dosage alpinia oxyphylla group increased significantly (P<0.05).(8) Effect on the content of ET in serumCompared with the model control group, the content of ET in serum of high dosage galangal group, high dosage grass cardamom group, high dosage fructus galangae group, low dosage fructus galangae group, low dosage galangal group, high dosage alpinia oxyphylla group, and low dosage alpinia oxyphylla group decreased significantly (P<0.01); the content of ET in serum of high dosage galanga group, low dosage galanga group, low dosage galangal group, high dosage alpinia oxyphylla group, and low dosage alpinia oxyphylla group decreased significantly (P<0.05).(9) Effect on the content of AchE in brain tissueCompared with the model control group, the content of AchE in brain tissue of high dosage galangal group, high dosage grass cardamom group, high dosage fructus galangae group increased significantly (P<0.01); the content of AchE in brain tissue of high dosage galanga group, low dosage galangal group, high dosage fructus galangae group increased significantly (P<0.05); the content of AchE in brain tissue of low dosage galanga group, low dosage grass cardamom group, low dosage fructus galangae group, and low dosage alpinia oxyphylla group had no significantly change (P>0.05).(10) Effect on the content of 5-HT in brainCompared with the model control group, the content of 5-HT in brain of high dosage galanga group, high dosage galangal group, high dosage grass cardamom group, high dosage fructus galangae group, low dosage fructus galangae group, high dosage alpinia oxyphylla group, low dosage alpinia oxyphylla group decreased significantly (P<0.01); the content of 5-HT in brain of low dosage grass cardamom group decreased significantly (P<0.05); the content of 5-HT in brain of the content of 5-HT in brain of and low dosage galangal group had no significantly change (P>0.05).(11) Effect on the content of NE in brainCompared with the model control group, the content of NE in brain of high dosage grass cardamom group, high dosage fructus galangae group, high dosage alpinia oxyphylla group decreased significantly (P<0.01); the content of NE in brain of low dosage fructus galangae group decreased significantly (P< 0.05); the content of NE in brain of high dosage galanga group, low dosage galanga group, high dosage galangal group, low dosage galangal group, low dosage grass cardamom group, high dosage alpinia oxyphylla group, and low dosage alpinia oxyphylla group had no significantly change (P>0.05).Coclusion:1. The best concentration of glacial acetic for establishing gastric ulcer rat model of cold syndrome is 15%.2. The cold syndrome of stomach ulcer induced by glacial acetic acid related with the increasing of the content of PGE2, EGF, TGF-α, NT, NO, AchE; and the decreasing of the content of IL-8, TNF-α, ET in blood serum,5-HT, and NE.3. The mechanism of treating stomach ulcer with cold syndrome of cold syndrome of may related with the function of reducing the amount of gastric juice secretion, hydrochloric acid in gastric juice, and pepsin activity; related with the function of increasing the content of PGE2, EGF, TGF-α, NT, NO, AchE, related with the function of decreasing the content of IL-8, ET in blood serum,5-HT, NE.4. Galanga, galangal, grass cardamom, fructus galangae, alpinia oxyphylla, which have genetic relationship, have the similar pharmacological effect on the function of warming stomach and dispelling cold. |
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