| ObjectiveGastroesophageal reflux disease(GERD)is a common disease of upper digestive tract characterized by reflux-related symptoms and(or)complications due to reflux of stomach contents(or gastroduodenal contents)into the esophagus.The recent prevalence of GERD has increased significantly,so it is the solid foundation for further improving the clinical efficacy of GERD by continuing to explore the pathogenesis on the basis of paying attention to its diagnosis and treatment.Chinese medicine has found that the syndrome of the stagnancy heat of the liver and stomach is a common syndrome of GERD during the clinical observation.However,owing to the limitations of the objective diagnostic evidences,the syndrome differentiation is still undefined to a certain extent.Therefore,this study aims to find the possible biological markers through metabolomics methods,by detecting plasma and urine samples of GERD patients and the syndrome of the stagnancy heat of the liver and stomach of GERD patients respectively,and analyzing its related metabolic material,and then to elucidate the possibly abnormal metabolic pathways in GERD and GERD patients with liver and stomach heat stagnation syndrome,and finally speculate the pathogenesis to provide the objective material basis for syndrome differentiation.This thesis is divided into two parts to discuss the related viewpoints and researches of GERD.The first part summarizes the literature reviews of TCM and Western medicine which includes two chapters.The first chapter has elaborated the TCM’s research status quo of GERD,focusing on the etiology,pathogenesis and distribution of syndromes.The second chapter has elaborated the mordern medicical progress of GERD,focusing on the etiology,pathogenesis and the status quo of metabolomics application in the study of Traditional Chinese Medicine.The second part is clinical trial,which is a metabolomics experimental study respectively tested with GERD and liver-stomach-stagnancy-heat pattern of GERD.The study tries to respectively find the possibly different biological markers in the process ofGERD.and the syndrome of the stagnancy heat of the.liver and stomach of GERD byanalyzing and discussing experiment results.And then to elucidate the possibly abnormal metabolic pathways in GERD and GERD patients with liver and stomach heat stagnation syndrome,and finally speculate the pathogenesis of GERD and GERD with the syndrome of the stagnancy heat of the liver and stomach.Methods(1)We collected blood and urine samples of 40 GERD patients(20 liver-stomach-stagnancy-heat syndrome,20 deficiency and qi reversal syndrome)and 35 healthy subjects respectively.Then we used 1H-NMR metabonomics technology to get corresponding metabolic spectra of the samples of GERD which was used as the experimental group while healthy volunteers as the control group.After the data were normalized and integrated,they were treated by SIMCA-P software to make principal component analysis(PCA)to give the principal component(PC).Then reuse PC for each set of samples metabolic spectra to make partial least squares-discriminant analysis(PLS-DA),orthogonal partial least squares-discriminant analysis(OPLS-DA)and response permutation testing(RPT)to get scatter plot(S-plot)and loading plot(L-plot),so as to intuitively show each set of samples metabolites group information.Then found the names and chemical shifts of differential metabolites,summarized and analyzed the trends of differential metabolite content,and finally identified the biological markers of GERD.(2)Using the above detection and analysis methods test the samples of GERD with liver-stomach-stagnancy-heat syndrome which was used as the experimental group while GERD with deficiency and qi reversal syndrome as the control group again,identified the biological markers of GERD with liver-stomach-stagnancy-heat syndrome.Results(1)40 patients in this study,female 30 cases,males 10 cases,minimum age 30 years,maxilmum age 79 years,mean age was 54.23±11.412 years.45～65-year-old patients account for 70%.(2)In addition to the main symptoms of acid reflux,heartburn,GERD patients show local symptoms of bloating,belching;burning sensation in stomach and stoimach pain,respectively accounting for 75%,67.5%,50%,45%,GERD withliver-stomach-stagnancy-heat syndrome mostly show bitter accounting for 27.5%while GERD with deficiency and qi reversal syndrome weariness accounting for 32.5%in systemic symptoms.(3)The mainly Inducing factors of GERD patients are temperament,irritation,and anxiety,respectively accounting for 62.5%,60.0%,50.0%.It is easier induced for GERD patients with liver-stomach-stagnancy-heat syndrome to eat spicy food accounting for 17.5%while GERD with deficiency and qi reversal syndrome to eat dessert and moody respectively accounting for 17.5%,15.0%.(4)Experimental results of plasma in the GERD experimental group and healthy control group showed:the content of τ-methylhistidine,N-methylhydantoin,levulinate in the GERD experimental group was higher than that in the healthy control group,and the difference was statistically significant(P<0.05);the content of isoleucine,valine,glucose,glutamine,glutamate,alanine,tyramine,lactate,leucine,LDL,VLDL,creatine phosphate was lower than that in the healthy control group,and the difference was statistically significant(P<0.05);(5)Experimental results of urine in the GERD experimental group and healthy control group showed:the content of trigonelline,1-methylnicotinamide,adenine in the GERD experimental group was higher than that in the healthy control group,and the difference was statistically significant(P<0.05);the content of choline,glucose,citrulline,glutamine,histidine,carnitine,tryptophan,lactate,cis-aconitate,glutamate,acetoacetate,creatinine,citrate,betaine,pyruvate,valine,arginine,glycine,2-aminobutyrate was lower than that in the healthy control group,and the difference was statistically significant(P<0.05);(6)Experimental results of plasma in the GERD with liver-stomach-stagnancy-heat syndrome group and GERD with deficiency and qi reversal syndrome group showed:the content of lactate,tyramine,τ-methylhistidine,creatine phosphate in the GERD with liver-stomach-stagnancy-heat syndrome group was higher than that in the GERD with deficiency and qi reversal syndrome group,and the difference was statistically significant(P<0.05);the content of glucose,valine,glutamine,glutamate,alanine,isoleucine,leucinewas lower than that in the GERD with deficiency and qi reversal syndrome group,and the difference was statistically significant(P<0.05).(7)Experimental results of urine in ’the GERD with liver-stomach-stagnancy-heat syndrome group and GERD with deficiency and qi reversal syndrome group showed:the content of lactate,citrulline,citrate,glutamine,pyruvate,succinate in the GERD with liver-stomach-stagnancy-heat syndrome group was higher than that in the GERD with deficiency and qi reversal syndrome group,and the difference was statistically significant(P<0.05);the content ofcreatinine,hippurate,glycine,glucose,tryptophan,choline was lower than that in the GERD with deficiency and qi reversal syndrome group,and the difference was statistically significant(P<0.05).Conclusion(1)In this study,the mean age of GERD patients was 54.23±11.412 years,which is consistent with the average age,40-60 years old,Chinese patients was diagnosed.45-65 year-old patients among them account for 70%,which is partly according with the current epidemiological findings.(2)In addition to the main symptoms of acid reflux,heartburn,GERD patients mostly show bloating,belching,burning sensation in stomach and stomach pain in local symptoms,GERD with liver-stomach-stagnancy-heat syndrome mostly show bitter while GERD with deficiency and qi reversal syndrome show weariness in systemic symptoms.(3)Emotional and dietary factors have an important influence on the pathogenesis of GERD This suggests that the patients should pay attention to the adjustment of mood and diet during daily life.(4)There are significant differences in plasma and urine metabolites between the GERD experimental group and the healthy group.This suggests that τ-methylhistidine,N-methylhydantoin,levulinate,isoleucine,valine,glucose,glutamine,glutamate,alanine,tyramine,lactate,leucine,LDL,VLDL,creatine phosphatein the plasma and trigonelline,1-methylnicotinamide,adenin,choline,glucose,citrulline;glutamine,histidine,carnitine,tryptophan,lactate,cis-aconitate,glutamate,acetoacetate,creatine,citrate,betaine,pyruvate,valine,arginine,glycine,2-aminobutyrate in the urine is related to the occurrence and development of GERD,and they may become biological markers of GERD.This may further explain that the occurrence and development of GERD may be related to the disorder of biochemical pathways such as energy metabolism,vitamin metabolism,nucleic acid metabolism,neurotransmitter metabolism,and oxidative stress.(5)There are some differences in plasma and urine metabolites between GERD with liver-stomach-stagnancy-heat syndrome group and.GERD with deficiency and qi reversal syndrome group.This suggests that lactate,tyramine,τ-methylhistidine,creatine phosphate,glucose,valine,glutamine,glutamate,alanine,isoleucine,leucine in the plasma and lactate,citrulline,citrate,glutamine,pyruvate,succinate,creatinine,hippurate,glycine,glucose,tryptophan,choline in the urine is related to the occurrence and development of GERD with liver-stomach-stagnancy-heat syndrome,and they may become biological markers of GERD.This may further explain that the occurrence and development of GERD with liver-stomach-stagnancy-heat syndrome may be related to the disorder of biochemical pathways such as energy metabolism,neurotransmitter metabolism and Gastrointestinal bacterial metabolism. |
PDF Full Text Request