| Objective: To investigate the therapeutic effect of Huangqi Jianzhong Decoction on gastric ulcer(GU)rats with deficiency cold of spleen and stomach and explore its mechanism.Methods:(1)24 male SD rats with body mass(160-180g)were randomly divided into normal group and spleen stomach deficiency cold type gastric ulcer model group(model group),12 rats in each group.The rat model of deficiency cold of spleen and stomach was established by filling stomach with centration of 1g/ml senna leaf and swimming at exhaustion for 9 days;from the 10 th day,by filling stomach with anhydrous ethanol and aspirin suspension for 4 days,in order to construct the rat model of deficiency cold of spleen and stomach.The body mass,food intake and water intake of rats in each group were measured,the anal temperature was measured the next day,and the mental state,hair,lips,toes color and tongue image of rats were closely observed.The rats were anesthetized and killed after modeling.The gastric tissues were taken and the morphological changes of gastric mucosa were observed by naked eye and HE staining.(2)60 SD male rats,body mass(160-180g),were randomly divided into normal group,spleen stomach deficiency cold type gastric ulcer model group(model group),model + esomeprazole group(positive control group)(esomeprazole,ESO),model + Huangqi Jianzhong Decoction low dose group(HQJZT,9.27 g / kg),model + Huangqi Jianzhong Decoction high dose group(HQJZT,18.54 g / kg),12 rats in each group.Rats in the normal group were given the same amount of distilled water,the other rats were given the preventive treatment.The positive control group was given 0.004g/kg esomeprazole,the low-dose and high-dose groups were given 9.27g/kg and 18.54g/kg Huangqi Jianzhong Decoction respectively,and the normal group and model group were given the same volume of distilled water by gavage for 4 consecutive days.The body mass,food intake and water intake of rats in each group were recorded.The anal temperature was measured the next day.The mental state,hair,nose,lip,toe color and tongue image of rats in each group were observed.At the end of the experiment,the rats were anesthetized and killed,take gastric tissue,observe the damage of gastric mucosa with naked eyes,count ulcer index and cure rate;take blood from abdominal aorta of rats,obtain serum by centrifugation,detect the content of IL-4,IL-10,No,TNF-?,PGE and MDA in serum of rats by ELISA method;detect the total acidity of gastric acid by sodium hydroxide dropping method;detect the activity of pepsin by ultraviolet spectrophotometry;take gastric tissue the expression of TLR-2 and My D88 related to the immune function of gastric mucosa was detected by fluorescence quantitative PCR,and the expression and distribution of TLR-2 and My D88 protein in gastric mucosa were detected by immunohistochemistry.Results:(1)Experimental results of the rat model of gastric ulcer of spleen stomach deficiency cold type(1)After the model of spleen and stomach deficiency cold,the mental state of rats in the model group was depressed,their hair was withered,they liked to pile up,they were tired and weak,their nose,lips and toes were white and lusterless,their tongue was dark purple,their stools were soft and occasionally loose.(2)Compared with the normal group,the body weight of the model group increased slowly(P < 0.01),the amount of food and water decreased(both P < 0.01),and the anal temperature decreased(P < 0.01).(3)In the model group,there were obvious erosions and bleeding spots on the gastric mucosa,covered with yellow pus on the surface;HE staining showed that the epithelial cells of gastric mucosa in the model group dropped off,and the submucosa appeared edema,bleeding and a large number of inflammatory cells infiltration.The above results indicate that the animal model of gastric ulcer of spleen stomach deficiency cold type has been successfully established.(2)Experimental results of therapeutic effect and mechanism of Huangqi Jianzhong Decoction on gastric ulcer of spleen stomach deficiency cold type in rats(1)Compared with the normal group,the mental state of the model group was depressed,the hair was withered,the color was yellowish,the back of bow was obvious,lazy,liked to pile up,and the response was slow;the lips,tongue and toenails were light white or dark purple;the stool was thin and soft,occasionally loose.After the treatment of ESO,low dose(9.27g/kg)and high dose(18.54g/kg)of HQJZT,the whole state of the rats was active,the hair was glossy,there was no reaction such as bow back and clumping,the lips,claw toes and tongue were light red or dark red,the stool was moderate soft and hard.(2)Compared with the normal group,the food intake and water intake of the model group decreased significantly(both P < 0.01),the body weight increased slowly(P < 0.01),and the anal temperature decreased significantly(P < 0.01).After the treatment of low-dose and high-dose HQJZT,the body weight,anal temperature,food intake and water intake of the HQJZT group increased significantly(P < 0.01).(3)The pathological changes of gastric mucosa in the model group were obvious,compared with the normal group,the ulcer index was significantly increased(P < 0.01).After the treatment of ESO,low dose and high dose HQJZT,the ulcer inhibition rate of high dose group of HQJZT was 79.04%,which was more obvious than that of other low dose group and ESO group.(4)The results of HE staining showed that compared with the normal group,the total scores of inflammatory cells,epithelial cells loss and bleeding in the model group were significantly increased(both P < 0.01);after the treatment of HQJZT,the damaged gastric mucosa was significantly improved,compared with the model group,the scores of inflammatory cells were significantly decreased(P < 0.01),epithelial cells loss and bleeding were decreased,and the total scores were significantly decreased(both P < 0.01).(5)Compared with the normal group,the total acidity of gastric acid and the activity of pepsin in the model group increased significantly(both P < 0.01).After the treatment of low dose and high dose of HQJZT,the total acidity of gastric acid and the activity of pepsin in the model group decreased significantly(both P < 0.01).(6)Compared with the normal group,NO,PGE,IL-4 and IL-10 in the serum of the model group decreased significantly(both P < 0.01),the levels of MDA and TNF-? increased significantly(both P < 0.01).After treatment with ESO,low and high dose of HQJZT,compared with the model group,the contents of No,PGE,IL-4 and IL-10 in serum of HQJZT rats increased significantly(both P < 0.01),and the contents of MDA and TNF-? decreased significantly(both P < 0.01).(7)Compared with the normal group,the expression of TLR-2 and My D88 genes closely related to mucosal immunity in the model group was significantly increased(both P < 0.01),after ESO,low and high dose of HQJZT treatment,compared with the model group,the expression of TLR-2 and My D88 genes in the gastric mucosa of the HQJZH group was significantly decreased(both P < 0.01).(8)Compared with the normal group,the expression of TLR-2 and My D88 protein in the upper cortex of the gastric mucosa of the model group increased significantly(both P < 0.01);After treatment with ESO,low and high dose of HQJZT,compared with the model group,the expression of TLR-2 and My D88 protein in the gastric mucosa decreased significantly(both P < 0.01).Conclusion: Huangqi jianzhong decoction has obvious therapeutic effect on GU rats of spleen stomach deficiency cold type.Its mechanism may be related to the inhibition of gastric acid secretion,the decrease of pepsin activity,the increase of mucosal protective factor content,the alleviation of oxidative lipid reaction of gastric mucosa,the alleviation of mucosal inflammatory reaction and the enhancement of gastric mucosal immune function by activating TLR-2 / My D88 pathway. |
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