| Objective: An animal model of chronic gastritis with spleen and stomach damp-h eat syndrome was established by endogenous damp-heat method,and the effect of Hu midity and heat resistance on γ-interferon(IFN-γ)secreted by T helper 1 cell(Th1)a nd interleukin-4(IL-4)secreted by T helper 2 cell and the protein expression of B-ly mphoma cell-2 gene(Bcl-2)and tumor suppressor gene(p53)in mice with spleen and stomach damp-heat syndrome was observed,to explore the method of Humidity and he at resistance on inflammatory factors and immune function in mice with spleen and st omach damp-heat syndrome.Methods: 80 SPF Kunming mice,half males and half females,were randomly divi ded into 6 groups for modeling;20 SPF Kunming mice,half males and half females, were used as the normal group.The model adopts the internal-caused damp-heat met hod,namely,high-fat and high-sugar diet(using liquor 10 m L/(Kg·d),oil 10g/(Kg·d)alternately for gavage every other day,and freely drinking honey and water)to replica te the animal model of spleen and stomach damp-heat syndrome.Then,the successfull y modeled mice were randomly numbered into groups,namely,the model group,the Huangqin talc decoction group,the low,medium,and high-dose groups of Shire Zhon gzhi Recipe(6.2,12.4,24.8 g/Kg),10 mice in each group;10 mice were randomly s elected from the normal group as the control group.Successfully modeled mice were given intragastric administration for 7 consecutive days,and resumed their normal diet.The control group and model group were given the same dose of normal saline intra gastrically.Two weeks later,after the last administration,fasting without water for 16 hours,blood was taken from the eyeballs,and the neck was sacrificed.The stomach t issue was taken,and half of it was fixed in 4% paraformaldehyde solution for more t han 24 hours,dehydrated,soaked in wax,embedded,3μm section and HE stained;The other half was frozen and stored in liquid nitrogen for WB experiment.Detect the c ontent of IFN-γ and IL-4 by ELISA;Use HE staining method to observe the changes of gastric tissue morphology;Western blotting was used to detect the protein expressi on levels of Bcl-2 and p53.Results:(1)Control group: The weight gradually increased,the reaction was sensiti ve,the food intake and water intake were normal,the stool was formed,and the hair was shiny.Model group: weight loss,lethargy,slow response,dry and dull hair,loos e stools,reduced food intake and water intake,and higher rectal temperature than the control group.Shire Zhongzhi Recipe and Huangqin Talc Decoction group: After one week of gavage,the food intake and water intake of the mice gradually increased,the ir mental state improved,their reaction was more sensitive than before,their weight in creased,the gloss of hair increased,and the rectal temperature decreased.Among them,the symptom improvement of the high-dose group of Shire Zhongzhi Recipe was the most obvious.(2)The serum IL-4 and IFN-γ values of mice in the model group were significantl y higher than those of the control group,and the IL-4 and IFN-γ values of the high and medium dose groups of Shire Zhongzhi Recipe were significantly lower than thos e of the model group.(3)The results of pathological tissue slices of mouse gastric mucosa showed that t he structure of the epithelial cells of the gastric mucosa of the control group was co mplete,the structure of the gastric pits were clear,and the epithelial cells were tightly arranged;The lamina propria is abundant in gastric glands,and the main cells and p arietal cells are normal in morphology and structure,and no obvious inflammation is s een.Compared with the control group,epithelial cells can be seen in the gastric muco sa in the model group;There is a decrease in gastric glands and hyperplasia of conne ctive tissue in the lamina propria;Edema can be seen in the submucosa,and the conn ective tissues are arranged loosely;A small amount of lymphocytes and neutrophils ca n be seen in the lamina propria and submucosa.Compared with the model group,the lamina propria of the positive drug group is rich in gastric glands,the principal cells and parietal cells are normal in morphology,edema is seen in the submucosa,the co nnective tissue is loosely arranged,and there is a small amount of lymphocyte infiltrat ion.With the increase of drug concentration in the low,medium and high dose group s of Shire Zhongzhi Recipe,the mucosal epithelial cell shedding was significantly red uced,and the infiltration of lymphocytes and neutrophils in the lamina propria and su bmucosa was significantly reduced.In the high-dose Shire Zhongzhi Recipe group,onl y a small amount of epithelial cells fell off in the mucosal layer,the lamina propria gastric glands were abundant,the main cells and parietal cells were normal in morpho logy,no obvious inflammation,and the gastric mucosa recovered most obviously.(4)Compared with the control group,the protein expression levels of Bcl-2 and p53 in the gastric tissue of the model group increased significantly;Compared with the model group,the protein expression of Bcl-2 and p53 in the low,medium and high d ose groups of Shire Zhongzhi Recipe was significantly reduced.Conclusion:(1)By suppressing the production of inflammatory factors IL-4 and IF N-γ,the self-made prescription Shire Zhongzhi Recipe,based on the principle of reduc ing hardship and pain,reduces inflammatory exudation and promotes the absorption of inflammation,thereby reducing tissue damage caused by excessive immune response.(2)The self-made prescription Shire Zhongzhi Recipe based on the principle of red ucing pain and suffering can inhibit the expression of Bcl-2 and p53 protein,promote the apoptosis of damaged cells,reduce the expression of carcinogens,and may have the effect of preventing cell canceration and repairing gastric mucosal damage.(3)The self-developed recipe for damp-heat Zhongzhu,which is based on the princ iple of relieving pain and suffering,is more complex,and its pharmacological effects need to be further studied.However,due to the limitation of experimental conditions and experimental funds,this experiment only reduces the mechanism of cell inflammat ory damage and apoptosis of the syndrome about chronic gastritis spleen and stomach damp-heat is preliminarily discussed,and the inherent shortcomings still need to be f urther improved and perfected. |
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