| Objective:This article firstly takes patients with reflux esophagitis of Stagnation-heat in Liver and Stomach as the research object to discuss the clinical efficacy and safety of Qingyuhejiang decoction;Then the rat model of reflux esophagitis was established.16 Sr RNA sequencing technique was used to study the relationship between reflux esophagitis and esophageal flora,and to observe the effect of Qingyuhejiangtang on esophagitis flora and related pathways in rats with reflux esophagitis,and to explore the mechanism of Qingyuhejiangtang,in order to provide new ideas and methods for clinical diagnosis and treatment of this disease.Methods:1.Theoretical discussion: This part reviews and summarizes the studies on the relationship between esophageal microflora and esophageal diseases,and expounds the important role of esophageal microflora in the occurrence and development of esophageal diseases.It provides a theoretical basis for further studying of the potential role of esophageal microflora in the physiological and pathological mechanism of esophageal diseases,exploring specific flora related to the pathogenesis of esophageal diseases and new ideas and methods for the treatment of esophageal diseases.This part systematically reviews and summarizes the ancient books of traditional Chinese medicine and modern scholars' understanding of the disease name,etiology,pathogenesis and treatment of reflux esophagitis,and expounds the effectiveness of traditional Chinese medicine in the treatment of reflux esophagitis.It provides a theoretical basis for further study on the mechanism of traditional Chinese medicine in the treatment of different syndrome types of reflux esophagitis.2.Clinical study: 94 patients with RE of Stagnation-heat in Liver and Stomach were randomly divided into western medicine group,traditional Chinese medicine group and combination group.The traditional Chinese medicine group was treated with Qingyuhejiang decoction,the western medicine group was treated with rabeprazole sodium enteric-coated capsule + mosapride citrate dispersible tablets,and the combination group was treated with Qingyuhejiang decoction + Rabeprazole sodium enteric-coated capsule + mosapride citrate dispersible tablets for 8 weeks,follow-up was performed 3months after withdrawal.Gerd Q scale score,TCM symptom score,comprehensive efficacy of TCM symptoms,effective rate of gastroscopy,anxiety score and long-term effect of the three groups before and after treatment were observed and compared,and the safety was evaluated before and after treatment.3.Experimental study: among 60 SD male rats,10 were randomly selected as sham operation group(group J),and the other 50 were used "anterior gastric ligation combined with external pyloric partial ligation" to establish the rat model of reflux esophagitis.Forty successful rat models were randomly divided into model group(group M),traditional Chinese medicine group(treated with Qingyuhejiang decoction)(group Z),western medicine group(treated with rabeprazole sodium enteric-coated capsule + mosapride citrate dispersible tablets)(group X)and combination group(treated with Qingyuhejiang decoction + rabeprazole sodium enteric-coated capsule + mosapride citrate dispersible tablets)(group ZX),there were 10 rats in each group.From the 15 th day after operation,the rats in the sham operation group(group J)and western medicine group(group X)were perfused with distilled water,and the other groups were given corresponding drugs for 14 days.16 Sr RNA sequencing technique was used to detect the esophageal flora of each group,and species composition analysis,Alpha(?)diversity analysis,Beta(?)diversity analysis,species difference and marker species analysis were carried out at the same time.At the same time,the pathological changes of rat esophagus were observed by hematoxylin-eosin(HE)staining,and the contents of lipopolysaccharide(LPS),interleukin(IL)-1?,interleukin(IL)-6and interleukin(IL)-8 in serum and esophageal tissue were detected by enzyme-linked immunosorbent assay(ELISA).The m RNA levels of Toll-like receptor 4(TLR4),nuclear factor kappa B(NF-?B)p65,Claudin-1(Cldn1)and Claudin-4(Cldn4)in esophageal tissues were detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),and the protein expressions of TLR4,NF-?B p65,Cldn1 and Cldn4 in esophageal tissues were detected by Western blotting(Western blot).Results:1.A total of 90 cases were completed in this study,including Chinese medicine group(n = 30),western medicine group(n = 30)and combination group(n = 30).First of all,we compared and analyzed the sex,age,course of disease,score of Gerd Q scale,single score of TCM symptoms,total score of TCM symptoms,total score of SAS and endoscopic grading of RE before treatment among the three groups.The results showed that there was no significant difference in these factors among the three groups(P>0.05).Then the comprehensive efficacy of TCM symptoms and gastroscopic efficacy of the three groups were observed.The comprehensive effective rates of TCM symptoms in traditional Chinese medicine group,western medicine group and combination group were 93.33%,90.00% and 96.67% respectively,and there was no significant difference between the groups(P>0.05).The effective rates under gastroscopy were 86.67%,93.33% and 96.67%,respectively.But there was no significant difference among the three groups(P>0.05).In terms of reducing the score of Gerd Q scale and the total score of TCM symptoms,the curative effect of the combination group was better than the traditional Chinese medicine group and the western medicine group(P<0.05),The curative effect of the traditional Chinese medicine group is similar to that of the western medicine group(P>0.05).In the treatment of main symptoms of TCM,the curative effect of western medicine group and combination group on heartburn was better than that of traditional Chinese medicine group(P<0.05).There was no significant difference in the efficacy of western medicine group and combination group on heartburn(P>0.05).There was no significant difference in the effect of acid regurgitation among the three groups(P>0.05).In the treatment of secondary symptoms of TCM,there was no significant difference in the curative effect of retrosternal pain,epigastric burning pain,abdominal distension and noisy hunger among the three groups(P > 0.05).The curative effect of belching in the combination group was better than that in the western medicine group(P<0.05).The curative effect of traditional Chinese medicine group and combination group on irritability,dry mouth and bitter mouth and constipation was better than that of western medicine group(P<0.05 or P<0.01).In terms of reducing the anxiety score,the curative effect of traditional Chinese medicine group and combination group was better than western medicine group(P<0.05),but there was no significant difference between traditional Chinese medicine group and combination group(P>0.05).In the long-term effect,there was no significant difference in the score of Gerd Q scale between the traditional Chinese medicine group and the combination group 3 months after treatment(P>0.05),but it was significantly higher in the western medicine group than after treatment(P<0.05),and the long-term effect of the traditional Chinese medicine group and combination group was better than western medicine group.During the 8 weeks of treatment and 3 months of follow-up,there were no adverse reactions in the three groups.2.There was no significant difference in the ? diversity index of the esophageal flora in each group(P>0.05).The PCo A analysis in ?-diversity analysis showed that the distribution of bacterial flora in the J group and the M group were clustered in different regions and were clearly separated;There was overlap,but all three treatment groups overlapped more with the J group and were closer to the J group.The Anosim test showed that there were significant differences in the composition of the overall comparative flora among the five groups(P<0.05);among them,there was a significant difference in the composition of the flora between the J group and M group(P<0.01);comparing the J group with the X group,there was a significant difference in the composition of the flora between them(P<0.05);comparing the J group with the Z group and ZX group,there was no significant difference between them(P >0.05);comparing the M group with the X group,Z group and ZX group,there were significant differences in the composition of microflora between them(P<0.05 or P<0.01);while there was no significant difference among the Z group,X group and the ZX group(P > 0.05).In terms of species composition,at the phylum level,the main taxa identified in each group were Firmicutes,Proteobacteria,Actinobacteria,Bacteroidetes,OD1,TM7 and so on.The relative abundance of Firmicutes in each group was in the following order:X group,ZX group,Z group,J group and M group(90.05%,83.14%,82.5%,74.60%,53.35%).The relative abundance of Proteobacteria in each group was in the following order: M group,J group,Z group,ZX group and X group(19.51%,10.99%,5.16%,3.55%,1.00%).The relative abundance of Actinobacteria in each group was in the following order: M group,X group,Z medicine group,ZX group,J group(9.70%,4.55%,2.95%,2.01%,1.24%).The relative abundance of Bacteroidetes in each group was in the following order: M group,X group,ZX group,J group,Z group(3.00%,1.54%,1.26%,0.43%,0.31%).In the J group,the relative abundance of Firmicutes was the highest,while the relative abundance of Firmicutes in the M group was significantly lower than that in the J group.the relative abundance of Firmicutes in the three treatment groups was significantly higher than that in the M group.The relative abundance of Proteobacteria,Actinobacteria and Bacteroidetes in the M group was significantly higher than that in the J group.After drug intervention,the relative abundance of Proteobacteria,Actinobacteria and Bacteroidetes in the three treatment groups was significantly lower than that in the M group.At the genus level,the main bacteria identified in each group were Lactobacillus,Streptococcus,Aggregatibacter,Bifidobacterium,Allobaculum,Clostridium and so on.The relative abundance of Lactobacillus in each group was in the following order:Z group,ZX group,X group,J group,M group(70.20%,63.43%,63.33%,49.80%,18.34%).The relative abundance of Streptococcus in each group was in the following order: J group,X group,ZX group,M group,Z group(21.08%,15.11%,13.25%,6.86%,4.58%).The relative abundance of Lactobacillus in the J group was the highest,but the relative abundance of Lactobacillus in the M group was lower.after drug intervention,the relative abundance of Lactobacillus in the three treatment groups was higher than that in the M group.Streptococcus was also dominant in the J group,but the relative abundance of Streptococcus in the M group was lower than that in the J group.After drug intervention,the relative abundance of Streptococcus in the X group and ZX group was higher than that in the M group.By LEf Se analysis,it was found that the marker species(biomarker)in the M group were Ochrobactrum,Allobaculum,Desulfovibrio,etc.,while the marker species in the J group,Z group,X group and ZX group were all Lactobacillus.3.Compared with the J group,the esophageal mucosal injury in the M group was the most serious,erosion or ulcer could be seen under naked eye and microscope,and a large number of inflammatory cells could be seen under microscope.Compared with the M group,there was no erosion and ulcer in the esophageal mucosa of rats in the Z group,X group and ZX group.Compared with the J group,the expression of LPS,IL-1 ?,IL-6 and IL-8 in serum and esophageal tissue of the M group was significantly increased(P<0.01).The expression of TLR4 and NF-? Bp65 m RNA and protein in esophageal tissue was significantly increased(P<0.01),while the expression of Cldn1,Cldn4 m RNA and protein in esophageal tissue was significantly decreased(P<0.01).Compared with the M group,the expression of LPS,IL-1 ?,IL-6 and IL-8 in serum and esophageal tissue in the three treatment groups decreased significantly(P<0.01),the expression of TLR4 and NF-? Bp65 m RNA and protein in esophageal tissue decreased significantly(P<0.01,P<0.05),and the expression of Cldn1,Cldn4 m RNA and protein in esophageal tissue increased significantly(P<0.01,P<0.05).Conclusions:1.Qingyuhejiang decoction can effectively treat reflux esophagitis with stagnant heat of liver and stomach,and cooperate with western medicine to improve the clinical efficacy,at the same time,it has better long-term efficacy and safety.2.The esophageal flora of rats with reflux esophagitis has changed: the main trend is the decrease of gram-positive bacteria and the increase of gram-negative bacteria,the marker species of esophageal flora in rats with reflux esophagitis were Ochrobactrum,Allobaculum and Desulfovibrio at the genus level.Qingyuhejiang decoction can regulate esophageal flora: after the intervention of Qingyuhejiang decoction,the number of Gram-positive bacteria increased and the number of Gram-negative bacteria decreased,and the marker species at the genus level was Lactobacillus.3.Qingyuhejiang decoction can inhibit LPS/TLR4/NF-?B signal pathway,reduce esophageal inflammation,enhance the function of esophageal mucosal barrier,promote esophageal mucosal repair,and effectively improve esophageal mucosal injury. |
PDF Full Text Request