Study On The Effect Of Huangqi Jianzhong Decoction On Gastric Tissue Injury And Key Molecules Of JAK2/STAT3 Pathway In Rats With Gastric Ulcer Of Spleen Stomach Deficiency Cold Type Based On "Disease Syndrome Prescription" Literature And Application Clues

Objective:This study first explored the corresponding relationship based on the literature clue of"disease syndrome prescription"between Huangqi Jianzhong Decoction and gastric ulcer（epigastric pain）of spleen stomach deficiency cold type,and confirmed the therapeutic effect of Huangqi Jianzhong Decoction on patients of spleen stomach deficiency cold type combined with modern clinical literature research;Then,based on JAK2/STAT3 signaling pathway,the possible molecular mechanism of Huangqi Jianzhong Decoction in effectively intervening the pathological damage of gastric tissue in rats with gastric ulcer of spleen stomach deficiency cold type was explored.Methods:Literature study:By using the method of traditional philology,this paper sorts out the relevant descriptions of stomachache in ancient medical books,systematically sorts out the disease name,etiology and pathogenesis,and treatment principles and methods,at the same time,it makes a study on the ancient literature of Huangqi Jianzhong Decoction from the evolution of prescription name and meaning,and discusses the theoretical basis of effective intervention of Huangqi Jianzhong Decoction on stomachache based on spleen stomach deficiency cold,At the same time,through the modern literature method,the clinical observation experiment of Huangqi Jianzhong Decoction in the treatment of gastric ulcer of spleen stomach deficiency cold type was systematically reviewed and analyzed.Experimental study:60 SPF healthy Wistar rats were randomly divided into blank control group and model group.The gastric ulcer rats with deficiency cold of spleen and stomach were modeled by comprehensive modeling method in model group,then gastric ulcer model rats were divided into five groups according to table of random numbers（n=10）:（1）model group,（2）positive control group,（3）Huangqi Jianzhong Decoction（high,medium and low）group(16g·kg^-1·d^-1/8g·kg^-1·d^-1/4g·kg^-1·d^-1).The rats from the control and model groups were given distilled water 10ml·kg^-1·d^-1 by intragastric administration,the positive control group was given 0.14g·kg^-1·d^-1 by intragastric administration,and the treatment lasted for 21 days.The general living condition and pathological changes of stomach tissue of rats were observed.The protein expression and phosphorylation level of JAK2,STAT3,HIF-1αand COX-2 in gastric tissue were detected by WB,immunohistochemical（IHC）was used to detect the expression of i NOS and e NOS protein in gastric tissue,enzyme linked immunosorbent assay（ELISA）was used to detect the contents of IL-10,IL-17,IL-6 and IL-13 in gastric tissue of rats in each group.Results:Literature study:（1）Epigastralgia was first known in Huangdi Neijing,and after Sui and Tang Dynasties,the name of epigastragaly was gradually established.With the development of the times,its treatment rules and prescription drugs were gradually improved.By the Ming and Qing Dynasties,the pathogenesis and prescription drugs were basically established;（2）Huangqi Jianzhong Decoction comes from synopsis of the golden chamber.In ancient times,many doctors emphasized its function of tonifying deficiency in the process of construction,and it was widely used in the treatment of various deficiency diseases.In modern times,it has been mainly used in the treatment of stomachache of spleen stomach deficiency cold type;（3）Systematic review of clinical observation literature shows that the treatment effect of Huangqi Jianzhong Decoction alone or Huangqi Jianzhong Decoction Combined with western medicine in the treatment of gastric ulcer of spleen stomach deficiency cold type is more significant than that of Western medicine alone,which is mainly reflected in relieving clinical symptoms,inhibiting Helicobacter pylori and reducing the recurrence rate.Experimental study:（1）General conditions:compared with the blank control group,rats in the model group were listless,curled up and huddled together with dishevelled hair,they showed obviously retarded reactions,abdominal distension,relatively slow increase in weight,less average daily food intake and significantly decrease in anal temperature,the difference is statistically significant（P<0.05）;Compared with model group,the symptoms of rats in each drug intervention group,such as thinning and lackluster hair,slow reaction,less food intake and activity,emaciation and drowsiness,were gradually alleviated with the increase of intervention time,the range of movement of the mice was getting normal,and the body mass growth was improved in varying degrees,the average daily food intake increased gradually,and the anal temperature increased in different degrees,especially in the high-dose and medium-dose groups of Huangqi Jianzhong Decoction,the difference is statistically significant（P<0.05）;（2）Pathological structure of gastric mucosa:（1）compared with the blank control group,ulcers were formed in the model group under 50 microscope,the epithelial layer and muscular layer disappeared at the ulcer site,the marginal epithelium of the ulcer was neat,and there was no obvious tendency of re-epithelialization.Compared with the model group,ulcers were formed in rats of the treatment groups,the epithelium and muscular layer disappeared,the tissues around the ulcers had an obvious tendency of re-epithelialization and began to migrate to cover the ulcers.（2）Compared with the blank control group,the rats in model group lost normal tissue structure under 100 microscope,inflammatory necrosis occured in local ulcer,the muscular layer was injured,almost all the epithelial cells were exfoliated and there was obvious infiltration in inflammatory cells.What can be seen obviously in each treatment group is that the normal epitheliums around the ulcers proliferated and migrated,covering the ulcer site in varying degrees,the inherent glands were mostly cystic dilatation,and inflammatory cells were scattered.The most significant pathological improvement was found in positive control group,high-dose and medium-dose groups of Huangqi Jianzhong Decoction;（3）Molecular biological detection:（1）The results of WB experiment showed that the protein expression and phosphorylation levels of JAK2,STAT3,HIF-1αand COX-2 in the gastric tissue of the model group were apparently higher than those of the blank control group,and the difference between the two groups was statistically significant（P<0.05）.Compared with the model group,the protein expression and phosphorylation levels of JAK2,STAT3,HIF-1αand COX-2 in the gastric tissues of rats in each treatment group were decreased to varying degrees,especially in the Huangqi Jianzhong Decoction high-dose group.The difference between the two groups was statistically significant（P<0.05）.（2）Immunohistochemical results showed that the protein expression levels of i NOS in the gastric tissue of rats in model group were significantly increased compared with the blank control group,and the protein expression level of e NOS was significantly decreased.The difference between the two groups was statistically significant（P<0.05）.Compared with model group,the protein expression levels of i NOS were down-regulated in all treatment groups,and expression of e NOS protein was up-regulated,especially in the high-dose group of Huangqi Jianzhong Decoction.The difference between the two groups was statistically significant（P<0.05）.（3）ELISA results showed that the contents of IL-10 and IL-13 in the stomach tissues of the rats with gastric ulcer of deficiency cold of spleen and stomach type were significantly decreased compared with the blank control group,while the contents of IL-17 and IL-6 were significantly increased.The difference between the two groups was statistically significant（P<0.05）.Compared with the model group,the contents of IL-10 and IL-13 in gastric tissue of rats in each treatment group were up-regulated and the contents of IL-17 and IL-6 were down-regulated in different degrees,especially in the high-dose group of Huangqi Jianzhong Decoction.The difference between the two groups was statistically significant（P<0.05）.Conclusions:Literature study:The name of epigastralgia was first found in Huangdi Neijing.The discussion of etiology,pathogenesis,treatment principle and treatment method of epigastralgia was gradually improved by physicians of past dynasties,and it was gradually completed in Ming and Qing Dynasties.Among them,Huangqi Jianzhong decoction,as the representative prescription of Jianzhong Buxu,is especially good at treating epigastric pain of spleen stomach deficiency cold type.For patients with gastric ulcer of spleen stomach deficiency cold type,Huangqi Jianzhong decoction has significant curative effect.Experimental study:Huangqi Jianzhong Decoction can significantly improve the general survival status and pathological changes of gastric mucosa in rats with spleen stomach deficiency cold type gastric ulcer.The specific mechanism may be that Huangqi Jianzhong Decoction can significantly reduce the expression of JAK2/STAT3 signaling pathway key molecules and downstream inflammatory and oxidative reaction related factors in gastric tissue of rats with spleen stomach deficiency cold type gastric ulcer.