Based On The CRF Pathway To Explore The Mechanism Of Sini San In The Treatment Of Functional Dyspepsia

Objective:Functional dyspepsia(FD)is one of the most common diseases in gastroenterology clinic.It is a functional gastrointestinal disease with the main clinical manifestations of fullness,early satiety,epigastric pain,epigastric burning sensation,etc.FD has a high incidence rate,but the clinical treatment benefits are relatively limited,which has obvious impact on the quality of life of patients,and also causes a large amount of medical resources to waste.Therefore,it is of great significance to carry out the relevant research of FD and further improve the clinical efficacy of FD.The pathogenesis of FD is relatively complex.From the current research situation,it is mainly related to abnormal gastric motility,impaired gastric receptivity,increased visceral sensitivity and abnormal duodenal function,especially the exploration of duodenal dysfunction is the focus of research at home and abroad.A large number of studies have shown that the damage of duodenal barrier function and the state of duodenal microinflammation are important components in the pathogenesis of FD.Among them,the dysfunction of duodenal barrier is closely related to the abnormal expression of TJs.However,the main changes of duodenal microinflammation are the number and degranulation of mast cells and eosinophils.When mast cells are activated,they will release histamine,tryptase and other bioactive substances,resulting in the increase of intestinal permeability.Psychological factors are independent risk factors for FD,and brain-gut axis is the key link.As one of brain gut peptides,corticotropin releasing hormone(CRF)is a key regulator in stress response,which can coordinate the body's feedback to stress stimulation through its specific receptor.CRF-R1 has a high affinity with CRF.It can increase intracellular cAMP concentration and activate protein kinase A(PKA)through G protein coupled receptor.It has been shown that CRF can activate mast cells and further induce intestinal epithelial barrier dysfunction by binding with CRF-R1 on epithelial mast cells under stress.Sini san is a traditional Chinese medicine formula,which has been passed on for thousands of years.It is also a commonly used medicine in the clinical treatment of FD.It can regulate the abnormal psychological state of patients and improve the quality of life.The results of previous studies showed that Sini san could restore the function of duodenal barrier in FD rats,but its specific mechanism was not cleared.The purpose of this study is to make a scientific evaluation on the clinical effect of Sini san in the treatment of FD by means of meta analysis,and to further explore the effect of Sini san on the function of duodenal barrier and the state of micro inflammation in FD rats through CRF pathway,so as to reveal its molecular mechanism.Methods:Part One:Meta analysis of Sini san in the treatment of functional dyspepsiaBy searching CNKI,Wanfang database,VIP,CBM,PubMed database,the Cochrane library database,web of science database,selected randomized controlled clinical studys on the treatment of functional dyspepsia with Sini san.Cochrane risk bias assessment tool was used to evaluate the quality of inclusion literature,and Review Manager 5.3 software was used to meta analyze the inclusion literature.Part Two:Study on the mechanism of Sini san in the treatment of FD through CRF pathway1.Effect of Sini san on gastric compliance and sensitivity in FD ratsThe model of FD rats was established by intragastric administration of iodoacetamide combined with tail clamping stress.The rats were randomly divided into normal group,model group and Sinisan treatment group.The gastric compliance and sensitivity of rats in each group were detected by electronic constant pressure apparatus and electrophysiological recorder.2.Effect of Sini san on duodenal barrier function in FD ratsThe model of FD rats was established by intragastric administration of iodoacetamide combined with tail clamping stress.The rats were randomly divided into normal group,model group and Sinisan treatment group.HE staining was used to observe the basic morphology of duodenum,using chamber to detect the transmembrane resistance(Teer),Western blot to detect the expression level of tight junction protein.The expression of ZO-1 and JAM-1 protein in duodenum of FD model group was significantly lower than that of normal group(P<0.01,P<0.01),while the expression of ZO-1 and JAM-1 protein in duodenum of Sini Powder treatment group was significantly higher than that of model group(P<0.01,P<0.05).3.Effect of Sini san on the activation of mast cells in FD ratsThe model of FD rats was established by intragastric administration of iodoacetamide combined with tail clamping stress.The rats were randomly divided into normal group,model group and Sinisan treatment group.The activation of mast cells(MC)in duodenum was observed by immunohistochemistry,the content of histamine in duodenum was detected by ELISA,and the expression of tryptase and PAR-2 mRNA was detected by qPCR.4.Effect of Sini san on CRF pathway in FD ratsThe model of FD rats was established by intragastric administration of iodoacetamide combined with tail clamping stress.The rats were randomly divided into normal group,model group and Sinisan treatment group.Western blot was used to detect the expression level of CRF protein in hypothalamus and duodenum.Immunohistochemistry was used to observe the immunopositive expression of CRF in spinal cord and duodenum.The expression of CRF-R1 mRNA was detected by qPCR.Results:Part One:Meta analysis of Sini san in the treatment of functional dyspepsia25 articles were included,including 2431 patients in Sini san group and 1263 patients in western medicine control group.The results of meta analysis showed that the total effective rate of Sini san treatment for FD was significantly higher than that of western medicine group(or=3.53;95%CI 2.78,4.47;P<0.00001);meanwhile,Sini san treatment also had significant advantages for FD patients with depression(or=3.33;95%CI 2.19,5.05;P<0.00001);Sini san can effectively alleviate the symptoms of upper abdominal pain and upper abdominal distention,and its curative effect is significantly better than that of western medicine group(or=3.85;95%CI 1.40,10.60;P=0.009 and or=2.99;95%CI 1.38,6.47;P=0.006);in terms of the improvement of Hamilton Depression Scale score,Sini san has a significant advantage over western medicine group(MD=-4.08;95%ci-4.82,-3.34;P<0.00001);in addition,the adverse reactions of Sini san were significantly less than that of Western Medicine(OR=0.14;95%CI 0.03,0.81;P=0.03).Part Two:Study on the mechanism of Sini san in the treatment of FD through CRF pathway1.Effect of Sini san on gastric compliance and sensitivity in FD ratsThe FD modeling method of iodoacetamide combined with tail clamping stress can reduce the gastric compliance of rats(20mmhg,P<0.05;40mmhg,P<0.01;60mmhg,P<0.01;80mmHg,P<0.01).Compared with the model group,the gastric compliance of rats in Sinisan treatment group increased significantly(20mmhg,P<0.05;40mmhg,P<0.01;60mmhg,P<0.01;80mmHg,P<0.01).Meanwhile,the gastric sensitivity of the model group was significantly higher than that of the normal group(at 20mmhg,40mmhg,60mmhg and 80mmHg,all P<0.01).Sini san treatment could significantly restore the gastric sensitivity of the rats(at 20mmhg,40mmhg,60mmhg and 80mmHg,all P<0.01)2.Effect of Sini san on duodenal barrier function in FD ratsHE staining showed that there was no obvious abnormality in the basic morphology of duodenum in each group.The duodenal transmembrane resistance(Teer)of FD model group was significantly lower than that of normal group(P<0.01),and after Sinisan treatment,the duodenal Teer of FD model group was significantly higher(P<0.01).3.Effect of Sini san on the activation of mast cells in FD ratsThe results of immunohistochemistry showed that the positive expression of the model group was significantly higher than that of the normal group(P<0.01);the positive expression of Sini san group was significantly lower than that of the model group(P<0.01).The results of ELISA showed that the content of duodenal histamine in the model group was significantly higher than that in the normal group(P<0.01),while the content of duodenal histamine in the FD rats treated with Sini san tended to be normal(P<0.01).The results of qPCR showed that the expression of tryptase and PAR-2 mRNA in duodenum of model group was significantly higher than that of normal group(P<0.01),while that of Sini san group was significantly lower(P<0.01)4.Effect of Sini san on CRF pathway in FD ratsWestern blot showed that the expression level of CRF protein in the hypothalamus and duodenum of the model group was significantly higher than that of the normal group(P<0.01);Sini san treatment could effectively reduce the expression level of CRF protein in the hypothalamus and duodenum of FD rats(P<0.01).The results of immunohistochemistry showed that the expression of CRF in spinal cord and duodenum of the model group was significantly higher than that of the normal group(P<0.01),while that in Sini san group was significantly improved(P<0.01).In addition,the expression level of CRF-R1 mRNA in duodenum of model group was significantly higher than that of normal group(P<0.01);Sini san treatment could significantly reduce the expression level of CRF-R1 mRNA in duodenum.Conclusion:1.The results of meta analysis:showed that Sini san has a better clinical effect in the treatment of FD,and has some advantages over western medicine in the aspects of overall symptoms,epigastric pain,epigastric distention,Hamilton Depression Scale score and adverse reactions.2.Experimental results show that Sini san can effectively improve the decrease of gastric compliance and the increase of gastric sensitivity in FD rats,alleviate the injury of duodenal mucosal barrier function and the state of micro inflammation in FD rats,and its mechanism may be related to the regulationof CRF/CRF-R1 signal pathway incentral nervous system and peripheral duodenum.