OBJECTIVETo investigate effects of electroacupuncture on gastric mucosal intestinal trefoil factor (ITF) gene expression in the rat of gastric mucosal injury induced by stress.

METHODSForty rats were randomly divided into 4 groups: blank group, model group, stomach channel group, gallbladder channel group. The normal group did not receive any processing, and the model group were fixed for 7 sessions and both the stomach channel group and the gallbladder channel group received 7 sessions of electro-acupuncture before modeling. The model was made by water restraint stress (WRS) for 10 hours. Index of gastric mucosal injury was detected and then the gastric mucosa tissue in each rat was taken and the expression of ITFmRNA of the tissue was detected by the reverse transcriptase-polymerase chain reaction (RT-PCR) method.

RESULTSThere was very significant or significant difference in the gastric mucosal injury as the stomach channel group and the gallbladder channel group compared with the model group (P < 0.05 or P < 0.01); electroacupuncture had a tendency of improving the expression of gastric mucosal ITFmRNA gene; the expression of ITFmRNA in the gallbladder channel group and the stomach channel group was significantly higher than that of the model group (P < 0.05, P < 0.01).

CONCLUSIONThe special regulation of electroacupuncture on gastric mucosal tissue is related with the release of epidermal growth factor and the expression of intestinal trefoil factor gene.

Animals ; Electroacupuncture ; Gastric Mucosa ; metabolism ; Gene Expression ; RNA, Messenger ; genetics ; Rats ; Stomach Diseases ; Stomach Ulcer ; metabolism

Tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) may be involved in the pathogenesis of peptic ulcers through suppression of fibrinolysis. This study was designed to investigate associations of t-PA and PAI-1 genes with clinical features of the patients with bleeding gastric ulcers. Eighty-four patients with peptic ulcers and 100 controls were studied between January 1998 and April 2000. We used polymerase chain reaction and endonuclease digestion to genotype for 4G/5G polymorphism in the promoter region of the PAI-1 gene and the Alurepeat insertion/deletion (I/D) polymorphism in intron h of the t-PA gene. Various clinical features, including lesion site, bleeding event, recurrence of ulcer, and rebleeding, were assessed using a multiple logistic regression model. The genotype distributions of both the t-PA and PAI-1 genes did not differ between the patient and control groups. The occurrence of the I/D or D/D genotype of t-PA was significantly higher in cases of duodenal ulcer (adjusted OR=4.39, 95% CI=1.12-17.21). When a dominant effect (i.e., 4G/4G or 4G/5G versus 5G/5G) of the 4G allele was assumed, the PAI-1 4G/4G genotype was independently associated with rebleeding after hemostasis (adjusted OR=5.07, 95% CI=1.03-24.87). Our data suggest that t-PA gene polymorphism is associated with duodenal ulcers, and that the PAI-1 gene may be a risk factor leading to recurrent bleeding after initial hemostasis.
Adult ; Aged ; Alu Elements/genetics ; DNA Mutational Analysis ; Duodenal Ulcer/complications ; Duodenal Ulcer/genetics* ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Human ; Male ; Middle Aged ; Mutagenesis, Insertional ; Peptic Ulcer Hemorrhage/etiology ; Peptic Ulcer Hemorrhage/genetics* ; Plasminogen Activator Inhibitor 1/genetics* ; Polymorphism (Genetics)* ; Promoter Regions (Genetics)/genetics ; Recurrence ; Sequence Deletion ; Stomach Ulcer/complications ; Stomach Ulcer/genetics* ; Tissue Plasminogen Activator/genetics*

This study aims to identify the active components and the mechanism of Jingqi Yukui Capsules(JQYK) in the treatment of gastric ulcer based on network pharmacology, and verify some key targets and signaling pathways through animal experiment. To be specific, first, the active components and targets of JQYK were retrieved from a Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM) and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets of gastric ulcer from GeneCards and Online Mendelian Inheritance in Man(OMIM) with the search term "gastric ulcer". The common targets of the two were the potential targets of the prescription for the treatment of the di-sease. Then, protein-protein interaction(PPI) network of key targets were constructed based on STRING and Cytoscape 3.7.2, followed by Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment by matescape database and pathway visualization by Omicshare. For the animal experiment, the improved method of Okabe was used to induce gastric ulcer in rats, and the model rats were classified into the model group, JQYK high-dose(JQYK-H), medium-dose(JQYK-M), and low-dose(JQYK-L) groups, Anweiyang Capsules(WYA) group, and Rabeprazole Sodium Enteric Capsules(RBPZ) group. Normal rats were included in the blank group. Rats in the blank group and model group were given distilled water and those in the administration groups received corresponding drugs. Then gastric ulcer healing in rats was observed. The changes of the gastric histomorphology in rats were evaluated based on hematoxylin-eosin(HE) staining, and the content of inducible nitric oxide synthase(iNOS) in rat gastric tissue was detected with Coomassie brilliant blue method. The mRNA and protein levels of some proteins in rat gastric tissue were determined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot(WB) to further validate some key targets and signaling pathways. A total of 206 active components and 535 targets of JQYK, 1 305 targets of gastric ulcer, and 166 common targets of the disease and the drug were yielded. According to PPI analysis and KEGG pathway enrichment analysis, multiple key targets, such as interleukin-6(IL-6), tumor necrosis factor(TNF), mitogen-activated protein kinase 1(MAPK1), MAPK3, and MAPK14, as well as nuclear factor kappa-B(NF-κB) signaling pathway, IL-17 signaling pathway, and leukocyte transendothelial migration in the top 20 key signaling pathways were closely related to inflammation. The key protein p38 MAPK and NF-κB signaling pathway were selected for further verification by animal experiment. The gastric ulcer in the JQYK-H group recovered nearly to the level in the blank group, with significant decrease in the content of iNOS in rat gastric tissue and significant reduction in the mRNA and phosphorylation levels of p38 MAPK and the mRNA and protein levels of NF-κB p65 in rat gastric tissue. The results indicated that JQYK can inhibit the phosphorylation of the key protein p38 MAPK and the expression of NF-κB p65 in the NF-κB signaling pathway, thereby exerting the anti-inflammatory effect and effectively improving the quality of gastric ulcer healing in rats. Thus, the animal experiment result verifies some predictions of network pharmacology.
Animal Experimentation ; Animals ; Capsules ; Gastric Mucosa/metabolism* ; Humans ; Network Pharmacology ; Rats ; Stomach Ulcer/genetics*

The vacuolated effect of Helicobacter (H. pylori) and its relationship to vacuolated cytotoxin antigen (VacA) were investigated by the method of cytotoxic test and SDS-pobyacrylamide gel electrophoresis (SDS-PAGE). Of the 62 clinical isolates, the broth culture filter (BCF) of 43 strains caused the Vero cell intracytoplasmically vacuolated. H. pylori strains were divided into H. pylori (Toxin+) group with vacuolated effect and H. pylori (Toxin-) group without vacuolated effect. The analysis of the BCF of H. pylori (Toxin+) and that of H. pylori (Toxin-) was studied by SDS-PAGE and Scan reader. A kind of protein with 87 ku molecular weight was recognized in the BCF of 30.23% (13/43) H. pylori (Toxin+) strains but in none of that of H. pylori (Toxin-) strains, the difference was statistically significant (P < 0.05). There was a significant and concordant relationship between OD of the protein band with 87 ku molecular weight and titer of vacuolated activity of H. pylori (Toxin+) (r = 0.67 and P < 0.05 by linear regression analysis). H. pylori strains were divided into H. pylori (Toxin+) group with vacuolated effect and H. pylori (Toxin-) group without vacuolated effect. The vacuolated effect of H. pylori (Toxin+) was caused by the protein with 87 ku molecular weight (VacA).
*Bacterial Proteins/genetics ; Genotype ; Helicobacter Infections/*microbiology ; Helicobacter pylori/*genetics ; Peptic Ulcer/*microbiology ; Stomach Diseases/microbiology ; Vacuoles

BACKGROUND: Determination of vacA mosaicism may be important because specific Helicobacter pylori vacA genotype can be used to predict different clinical outcome. The aim of this study was to assess the relationship of vacA genotypes of Helicobacter pylori to cagA status and its development of peptic ulcer diseases in Korean patients. METHODS: Gastric biopsy specimens were obtained from 53 patients with gastric ulcer(GU), 57 with duodenal ulcer (DU) and 26 with chronic gastritis(CG) patients; all patients were infected with Helicobacter pylori. Bacterial mRNAs in the gastric mucosa were amplified by RT-PCR, using synthetic oligonucleotide primers specific for the vacA and the cagA gene. Patients with vacA s1 subtype were further examined to determine whether they had s1a or s1b subtype. RESULTS: There was no correlation in frequency of vacA s1 and/or s1a genotype between CG and either GU or DU, as the vacA s1 and s1a/m1 were present in the majority of strains independent of clinical status(s1 ; 100.0% versus 94.3 % or 93.0 % and s1a/m1 ; 76.9% versus 62.3% or 64.9%, res pectively). Likewise, there was no difference in the prevalence of the cagA gene between CG and either GU or DU patients (92.3% versus 90.6% or 98.2%, respectively). In addition, the cagA-negative status did not predict the presence of vacA s2 genotype. CONCLUSION: These results strongly suggest that either cagA or vacA s1 and/or s1a is not proved to be a useful marker to distinguish disease-specific Helicobacter pylori strains for the development of peptic ulcer diseases in Korean patients.
Adolescence ; Adult ; Aged ; Aged, 80 and over ; Bacterial Proteins/analysis* ; Base Sequence ; Biopsy, Needle ; Chi-Square Distribution ; Chronic Disease ; Duodenal Ulcer/pathology ; Duodenal Ulcer/genetics ; Female ; Gastritis/pathology ; Gastritis/genetics ; Genotype ; Helicobacter Infections/pathology ; Helicobacter Infections/genetics* ; Helicobacter pylori/genetics* ; Human ; Korea ; Male ; Middle Age ; Molecular Sequence Data ; Peptic Ulcer/pathology ; Peptic Ulcer/genetics* ; Polymerase Chain Reaction ; Probability ; Prognosis ; Sensitivity and Specificity ; Stomach Ulcer/pathology ; Stomach Ulcer/genetics ; Support, Non-U.S. Gov't ; Tissue Culture

The vacuolated effect of Helicobacter (H. pylori) and its relationship to vacuolated cytotoxin antigen (VacA) were investigated by the method of cytotoxic test and SDS-pobyacrylamide gel electrophoresis (SDS-PAGE). Of the 62 clinical isolates, the broth culture filter (BCF) of 43 strains caused the Vero cell intracytoplasmically vacuolated. H. pylori strains were divided into H. pylori (Toxin+) group with vacuolated effect and H. pylori (Toxin-) group without vacuolated effect. The analysis of the BCF of H. pylori (Toxin+) and that of H. pylori (Toxin-) was studied by SDS-PAGE and Scan reader. A kind of protein with 87 ku molecular weight was recognized in the BCF of 30.23% (13/43) H. pylori (Toxin+) strains but in none of that of H. pylori (Toxin-) strains, the difference was statistically significant (P < 0.05). There was a significant and concordant relationship between OD of the protein band with 87 ku molecular weight and titer of vacuolated activity of H. pylori (Toxin+) (r = 0.67 and P < 0.05 by linear regression analysis). H. pylori strains were divided into H. pylori (Toxin+) group with vacuolated effect and H. pylori (Toxin-) group without vacuolated effect. The vacuolated effect of H. pylori (Toxin+) was caused by the protein with 87 ku molecular weight (VacA).
Adolescent ; Adult ; Bacterial Proteins ; genetics ; Female ; Genotype ; Helicobacter Infections ; microbiology ; Helicobacter pylori ; genetics ; Humans ; Male ; Middle Aged ; Peptic Ulcer ; microbiology ; Stomach Diseases ; microbiology ; Vacuoles

OBJECTIVETo explore relevant material basis of moxibustion for recovering gastric mucosal lesion. METHODL Forty-five SD rats were randomly divided into a normal goup, a model group, an acupoint group and a control group, 15 rats in the model group and 10 rats in the rest three groups. Except the normal group, binding and cold stress method were used to establish gastric mucosa injury model. The suspended moxibustion was applied in the acupoint group and control group at acupoints of the stomach meridian ("Liangmen" (ST 21) and "Zusanli" (ST36) and control acupoints (Laterally 1cm next to the "Liangmen" (ST 21) and Zusanli" (ST36), once a day, consectutively for 12 days. After 12 days, morphology of gastric mucosal was observed under optical microscope; protein fingerprints of gastric mucosa cell in rats were detected by protein fingerprint technology, weak cation chip and weak anion chip. Also mass to charge ratio of differential proteins in groups were compared and analyzed.

RESULTSCompared with the model group, index of gastric mucosal lesion in the acupoint group was reduced and its morphology was obviously improved (P<0.05). Campared with control group, index and morphology of gastric mucosal lesion were significantly improved in the acupoint group (P<0.05). According to test of weak cation chip, there was four marker proteins that had expression differences, indicating moxibustion at acupoints of stomach meridian could inrease expression of three marker protein whose molecular weight was 1354Da, 5692Da and 8432Da (all P<0.05) while reduce expression of marker protein with molecular weight of 3287Da (_<0.05). According to test of weak anion chip, moxibustion at acupoints of stomach meridian could increase expression of three marker proteins whose molecular weight was 2412 Da, 3026Da and 6475 Da (allP<0.05).

CONCLUSIONMoxibustion at acupoints of the stomach meridian could regulate differential expression of gastric mucosa cell-related marker protein in rats with acute gastric ulcer and recover gastric mucosal lesion, it's effect is better than that of the points of laterally 1 cm next to acupoint.

Acupuncture Points ; Animals ; Female ; Gastric Mucosa ; metabolism ; Humans ; Male ; Moxibustion ; Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stomach Ulcer ; genetics ; metabolism ; therapy

OBJECTIVETo observe the inflammatory reaction, nuclear factor-kappaB (NF-kappaB) mRNA and protein expression in stomach tissue of rats with gastric ulcer recurrence and the effect of Jianwei Yuyang granule (JYG) on them.

METHODSGastric ulcer and its recurrent lesion were successively induced by acetic acid and interliukin1-beta (IL-1beta), and the model rats were divided into the sham operation group, the model group, the omeprazole (correction of omepraxole) group and the JYG group to observe the state of chronic inflammatory cell, neutrophil count, NF-kappaBmRNA and protein expression in stomach tissue.

RESULTSOn the 16th and 92th day after administration, the increase of chronic inflammatory cell, neutrophil, NF-kappaBmRNA and protein expression in the model group was more significant than those in the sham operated group (P < 0.01), while that was lower in the JYG group than in the model group (P < 0.05, P <0.01), but with no remarkable difference to the omepraxole group. In addition, the mRNA and protein expression of NF-kappaB were correlated closely with the count of chronic inflammatory cell and neutrophil respectively (P < 0.01).

CONCLUSIONNF-kappaB may play an important role in regulating inflammatory reaction during the healing and recurrence processes of gastric ulcer induced by acetic acid. JYG may suppress inflammatory reaction by inhibiting the activation and expression of NF-kappaB in stomach tissue, which may be one of the mechanisms of JYG in preventing the recurrence of gastric ulcer.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gene Expression ; Male ; NF-kappa B ; biosynthesis ; genetics ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recurrence ; Stomach ; metabolism ; Stomach Ulcer ; drug therapy ; metabolism

Recent evidence suggests that gastric mucosal injury induces adaptive changes in DNA methylation. In this study, the methylation status of the key tissue-specific genes in normal gastric mucosa of healthy individuals and cancer patients was evaluated. The methylation-variable sites of 14 genes, including ulcer-healing genes (TFF1, TFF2, CDH1, and PPARG), were chosen from the CpG-island margins or non-island CpGs near the transcription start sites. The healthy individuals as well as the normal gastric mucosa of 23 ulcer, 21 non-invasive cancer, and 53 cancer patients were examined by semiquantitative methylation-specific polymerase chain reaction (PCR) analysis. The ulcer-healing genes were concurrently methylated with other genes depending on the presence or absence of CpG-islands in the normal mucosa of healthy individuals. Both the TFF2 and PPARG genes were frequently undermethylated in ulcer patients. The over- or intermediate-methylated TFF2 and undermethylated PPARG genes was more common in stage-1 cancer patients (71%) than in healthy individuals (10%; odds ratio [OR], 21.9) and non-invasive cancer patients (21%; OR, 8.9). The TFF2-PPARG methylation pattern of cancer patients was stronger in the older-age group (> or =55 yr; OR, 43.6). These results suggest that the combined methylation pattern of ulcer-healing genes serves as a sensitive marker for predicting cancer-prone gastric mucosa.
Biological Markers/metabolism ; Cadherins/genetics ; CpG Islands ; *DNA Methylation ; Female ; *Gastric Mucosa/pathology/physiology ; Gene Expression Regulation, Neoplastic ; Growth Substances/genetics ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; PPAR gamma/genetics ; Peptides/genetics ; *Stomach Neoplasms/genetics/pathology ; *Stomach Ulcer/genetics/pathology ; Tumor Suppressor Proteins/genetics ; Wound Healing/*genetics

OBJECTIVE: To observe the effect of acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) on intestinal flora and Toll-like receptors-4 (TLR4) in brain and intestinal tissue in rats with stress gastric ulcer (SGU), and to explore the possible mechanism of acupuncture for SGU. METHODS: Thirty-one male SD rats were randomly divided into a blank group ( RESULTS: Compared with the blank group, the gastric mucosal damage index was significantly increased in the model group ( CONCLUSION Acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) could alleviate SGU in rats, and its mechanism may be related to increasing the diversity of intestinal flora, promoting the disorder of intestinal flora to normal, and reducing the overexpression of TLR4 in brain and intestinal tissues.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Brain ; Gastrointestinal Microbiome ; Male ; Rats ; Rats, Sprague-Dawley ; Stomach Ulcer/therapy* ; Toll-Like Receptor 4/genetics*