Clinical Observation Of Tiaogan Jiedu Huoxue Decoction In Treating Chronic Atrophic Gastritis And JAK2/STAT3 Pathway Intervention Research

Part One: Clinical Observation of Tiaogan Jiedu Huoxue Decoction in Treating Chronic Atrophic GastritisObjective: To observe the clinical efficacy of Tiaogan Huoxue Jiedu decoction on patients with Chronic atrophic gastritis(CAG).Methods: A total of 66 CAG patients with liver depression and blood stasis toxin were randomly divided into treatment group and control group.The treatment group was given Tiaogan Jiedu Huoxue prescription granules,1bag once,twice a day.The control group was given Morodan concentrated pills,1 bag at a time,3 times a day.The course of treatment was 24 weeks,and the changes in the scores of TCM syndromes,gastroscopic manifestations and pathological visual simulation scores before and after treatment in the two groups were recorded in detail.The changes of serum pepsinogen(PG)PGI,PGII,and gastrin-17(G-17)before and after treatment were observed.Results:1.Analysis of clinical curative effect: The patients in the treatment group have significantly reduced TCM symptom scores compared with those before treatment,and they are better than the control group in improving epigastric burning pain,hypochondriac pain,dry mouth,bitter mouth,and dry stool(P<0.05);the control group had significantly lower TCM symptom scores compared with those before treatment,and the effect of improving the symptoms of gastric filling was better than that of the treatment group(P<0.05);there was no significant difference in other scores between the treatment group and the control group(P>0.05).2.Gastroscopy: After treatment,the scores of erythema,atrophy,erosion and intramucosal hemorrhage in the treatment group under gastroscopy were improved,and the erythema and atrophy of the control group were improved;the treatment group was better than the control group in terms of obvious improvement in erythema and atrophy(P< 0.05).There was no significant improvement in erosion and intramucosal hemorrhage between the two groups(P>0.05),and there was no significant difference between the two groups(P>0.05).3.Pathological aspects: After treatment,the treatment group is better than the control group in improving gastric mucosal pathological atrophy and chronic inflammation scores(P<0.05).There was no significant difference between the treatment group and the control group in the scores of pathological intestinal metaplasia,dysplasia and activity of gastric mucosa(P>0.05).4.Serological index analysis: After treatment,the levels of PGI and PGR in the two groups were higher than those before treatment(P<0.05),and the levels of PGII and G-17 in the two groups after treatment were lower than those before treatment,and the treatment group was superior to the control group(P<0.05);Summary:Tiaogan Jiedu Huoxue decoction can improve the clinical symptoms,gastroscopic pathological state and abnormal serological indicators of CAG patients,and has a good effect in the treatment of CAG.Part two: Effect of Tiaogan Jiedu Huoxue Recipe on Chronic Atrophic Gastritis Model RatsObjective: In this experiment,a CAG rat model was established by the N-methyl-N’-nitro-nitrosoguanidine(MNNG)compound method to observe the effect of Tiaogan Jiedu Huoxue decoction on the general condition of CAG rats,and the therapeutic effect of Tiaogan jiedu Huoxue decoction on glandular atrophy and intestinal metaplasia in CAG rats.Methods: After 1 week of adaptive feeding,ninety 8-week-old SPF male SD rats weighing 200±20 g were randomly divided into 2 groups,namely:blank group(BC)with 10 and CAG model group(MC)with 80.The blank group was given a normal diet and free drinking water.CAG model group was given 180ug/m L MNNG solution for free drinking combined with sodium salicylate gavage and hunger-satisfaction diet and tail-clamping method to replicate the CAG rat model.The model was continuously established until the14 th week,the pathology of two randomly selected rats both showed atrophy of proper gastric mucosal glands and intestinal metaplasia,indicating that the model was successful.After successful modeling,the CAG model group was randomly divided into 5 groups: model group,Tiaogan Jiedu Huoxue Decoction high-dose group,Tiaogan Jiedu Huoxue Decoction medium-dose group,Tiaogan Jiedu Huoxue Decoction low-dose group,and Morodan group.The normal control group and the model control group were given saline intragastrically.The intragastric administration concentrations of Tiaogan Jiedu Huoxue decoction in high,medium and low dose groups were27.72g/kg/d,13.86g/kg/d,6.93g/kg/d,respectively.The dosage of Morodan group was 0.50g/kg/d.The gavage volume is 10ml/kg,once a day,for 10 consecutive weeks.After the experiment,the gastric mucosa from the gastric antrum was taken to prepare gastric mucosal epithelial tissue sections.HE staining and AB-PAS staining were used to observe the gastric mucosal epithelial atrophy and intestinal metaplasia of rats in each group.Results:1.HE staining showed that the gastric mucosal epithelial cells of the blank group were neatly arranged,the glands were regular in shape,no inflammatory cell infiltration was seen,and the gastric mucosa was thick.In the model group,most of the gastric mucosal glands disappeared,the mucosal muscle layer was thickened,inflammatory cells infiltrated in the interstitium,some lymphoid follicles were formed,and intestinal metaplasia was seen.There are varying degrees of degeneration or necrosis of exfoliated cells,thinning of the mucosa,disordered arrangement of gland cells,cystic expansion of the glands,different sizes of epithelial cells,interstitial edema and hyperemia.Intestinal metaplasia appeared in the entire gastric mucosa,and a large number of goblet cells were seen.The gastric mucosa of rats in the Morodan group was pink,with poor gloss,thinning of the gastric mucosa,disordered folds,and infiltration of a large number of inflammatory cells.Interstitial edema,hyperemia,inflammatory cell infiltration,and a large number of intestinal metaplasias in the regional mucosa.The pathology of the three dose groups of Tiaogan Jiedu Huoxue Decoction is between that of Morodan and the blank group.Among them,the pathological improvement of the high-dose Tiaogan Jiedu Huoxue Decoction group is the most obvious,followed by the middle-dose Tiaogan Jiedu Huoxue Decoction group,followed by Tiaogan Jiedu Huoxue decoction.The pathological improvement effect of the low-dose prescription group is not obvious.2.AB-PAS staining showed that the gastric mucosal epithelium of the blank group had no blue intestinal metaplasia;the model group had diffuse intestinal metaplasia.After treatment with Tiaogan Jiedu Huoxue decoction,the range of intestinal metaplasia was significantly reduced.The high-dose and middle-dose groups of Tiaogan Jiedu Huoxue decoction improved most significantly,while the low-dose and Morodan groups did not improve significantly.Summary:Tiaogan Jiedu Huoxue Decoction can improve the pathological changes of CAG rats and delay the development of atrophy and intestinal metaplasia.Part three: Observation on the therapeutic effect of Tiaogan Jiedu Huoxue Decoction on the intervention of chronic atrophic gastritis in rats based on JAK2/STAT3 pathwayObjective: To observe the expression of JAK2/STAT3 signaling pathway in the CAG rat model,and to explore the regulatory effect of Tiaogan Jiedu Huoxue decoction on the key factors of JAK2/STAT3 signaling pathway.Methods: 1.Elisa method to detect IL-6 and TNF-α expression;2.Real-time PCR to detect JAK2 and STAT3 m RNA expression level;3.Western Blot method to detect JAK2,p-JAK2,STAT3,p-STAT3 protein in rat gastric mucosa Expression;4.Immunohistochemical method to detect Bax,Bcl-x L,Bcl-2 m RNA expression in rat gastric mucosa tissue;5.TUNEL staining to detect DNA damage.Results:1.Elisa method to detect the expression of serum IL-6 and TNF-αCompared with the blank group: the expression of serum IL-6 and TNF-αin the model group increased significantly(P<0.05).Compared with the model group: the expression of IL-6 and TNF-α in serum of rats in each dose group of Tiaogan Jiedu Huoxue decoction and Morodan group decreased significantly(P<0.05).2.Real-time PCR detection of JAK2 and STAT3 m RNA expression levels in the gastric mucosa of each group of rats JAK2,STAT3 m RNA amplification curve is smooth,the dissolution curve shows a sharp single peak,indicating that each group of rats gastric mucosal RNA amplification and dissolution effect is better.Compared with the blank group: the expression levels of JAK2 and STAT3 m RNA in the gastric mucosa of the model group increased significantly(P<0.05).Compared with the model group: Tiaogan Jiedu Huoxue decoction groups and Morodan group intervention significantly decreased STAT3 m RNA expression in gastric mucosa of CAG rats(P<0.05),but the decrease in JAK2 m RNA expression level was not statistically significant(P>0.05).3.Western Blot method to detect JAK2,p-JAK2,STAT3,p-STAT3 protein expression in gastric mucosa tissueCompared with the normal group,the values of p-JAK2/JAK2 and p-STAT3/STAT3 in the gastric tissue of the model group increased(P<0.05).Compared with the model group,the values of p-JAK2/JAK2 and p-STAT3/STAT3 in each dose group of Tiaogan Jiedu Huoxue decoction and Morodan group were decreased(P<0.05).4.Immunohistochemical method to detect the expression of Bax,Bcl-x L,and Bcl-2 m RNA in gastric mucosal tissueCompared with the blank group,the expression level of Bax m RNA in the gastric tissue of the model group increased,and the expression level of Bcl-2 and Bcl-xl m RNA decreased(P<0.05).Compared with the model group,the expression levels of Bax m RNA in the Tiaogan Jiedu Decoction groups and the Morodan group decreased to varying degrees,and the m RNA expression levels of Bcl-2 and Bcl-xl increased in different degrees(P<0.05).5.TUNEL positive detection of rat gastric cells in each groupCompared with the BC group,the TUNEL positive cells in the MC group were significantly increased(P<0.05);compared with the MC group,the TUNEL positive cells in the TH,TM,and TL groups and the DC group were significantly reduced(P<0.05).Summary: Tiaogan Jiedu Huoxue decoction may block or even reverse the transition from chronic atrophic gastritis to gastric cancer by regulating the abnormal state of JAK2/STAT3 signaling pathway and related factors.Conclusions:1.Tiaogan Jiedu Huoxue decoction can improve the clinical symptoms,gastroscopic pathological state and abnormal serological indicators of CAG patients,and has a good effect in the treatment of CAG.2.Tiaogan Jiedu Huoxue decoction can improve the pathological changes of CAG rats and delay the development of atrophy and intestinal metaplasia.3.Tiaogan Jiedu Huoxue decoction may block or even reverse the transition from chronic atrophic gastritis to gastric cancer by regulating the abnormal state of JAK2/STAT3 signaling pathway and related factors.