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*

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*

OBJECTIVETo explore the analgesic effect and action mechanism of electroacupuncture (EA) on gastric ulcer rats with liver-depression syndrome.

METHODSThrough open-field experimental method, 45 qualified SPF-grade male SD rats were selected and divided into a blank group, a model group and an EA group according to random number table method, 15 rats in each group. The model of gastric ulcer rats with liver-depression syndrome was established in the model group and the EA group by using chronic unpredictable stimulation combined with acetic acid burning method. Rats in the blank group did not receive intervention. Rats in the model group were treated with fixation and immobilization for 13 days. Rats in the EA group were treated with EA at "Liangqiu" (ST 34) and "Ganshu" (BL 18); EA voltage was 2 V; disperse-dense wave was selected with 4 Hz of disperse wave and 15 Hz of dense wave, and the intensity of EA was according to the slight vibration of local skin and; muscles; the needles were retained for 20 min, once a day for consecutive 6 days; there was an interval of 1 day' and the treatment was given for 2 weeks. The general condition, open-field experimental result and gastric ulcer index were observed; the western blotting method was applied to measure the expression of vanilloid receptor subtype 1 (VR1) in hypothalamus and gastric antral mucosal, and ELISA method was applied to test the expression of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) in hippocampus.

RESULTSAfter model establishment, the general behavior condition in the model group was inferior to that in the blank group, which was obviously improved after EA. The range of motion in the model group was less than that in the blank group (P<0.01) while that in the EA group was higher than that in the model group (P<0.01). The ulcer inhibition rate was. 54.95%, and the ulcer index in the EA group was lower than that in the model group (P<0.01). Compared with; the blank group, the expression of VR1 in hypothalamus and gastric antral mucosal in the model group was increased (P<0.05); compared with the model group, the expression of VR1 in the EA group was reduced (P<0.05). Compared with the blank group, the expression of 5-HT an NE in hippocampus in the model group was significantly reduced (both P<0.01); compared with the model group, the expression of 5-HT and NE in the EA group was increased (both P<0.01).

CONCLUSIONEA at "Liangqiu" (ST 34) and "Ganshu" (BL 18) has certain analgesic effect in gastric ulcer rats with liver-depression syndrome, which is likely to be related with lowering the contents of VR1 in hypothalamus and gastric antral mucosal and increasing the content of 5-HT and NE in hippocampus.

Acupuncture Analgesia ; Acupuncture Points ; Animals ; Electroacupuncture ; Humans ; Liver ; physiopathology ; Male ; Nociceptive Pain ; genetics ; metabolism ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism ; Stomach Ulcer ; genetics ; metabolism ; physiopathology ; therapy ; TRPV Cation Channels ; genetics ; metabolism

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

TNF-alpha is a major cytokine involved in inflammatory bowel disease (IBD). In this study, water extract of Grifola frondosa (GFW) was evaluated for its protective effects against colon inflammation through the modulation of TNF-alpha action. In coculture of HT-29 human colon cancer cells with U937 human monocytic cells, TNF-alpha-induced monocyte adhesion to HT-29 cells was significantly suppressed by GFW (10, 50, 100 microg/ml). The reduced adhesion by GFW correlated with the suppressed expression of MCP-1 and IL-8, the major IBD-associated chemokines. In addition, treatment with GFW significantly suppressed TNF-alpha-induced reactive oxygen species production and NF-kappaB transcriptional activity in HT-29 cells. In differentiated U937 monocytic cells, LPS-induced TNF-alpha production, which is known to be mediated through NF-kappaB activation, was significantly suppressed by GFW. In an in vivo rat model of IBD, oral administration of GFW for 5 days (1 g/kg per day) significantly inhibited the trinitrobenzene sulfonic acid (TNBS)-induced weight loss, colon ulceration, myeloperoxidase activity, and TNF-alpha expression in the colon tissue. Moreover, the effect of GFW was similar to that of intra-peritoneal injection of 5-aminosalicylic acid (5-ASA), an active metabolite of sulfasalazine, commonly used drug for the treatment of IBD. The results suggest that GFW ameliorates colon inflammation by suppressing production of TNF-alpha as well as its signaling through NF-kappaB leading to the expression of inflammatory chemokines, MCP-1 and IL-8. Taken together, the results strongly suggest GFW is a valuable medicinal food for IBD treatment, and thus may be used as an alternative medicine for IBD.
Animals ; Cell Adhesion/drug effects/immunology ; Cell Extracts/administration & dosage/*pharmacology ; Chemokine CCL2/biosynthesis/genetics ; Coculture Techniques ; Colon/drug effects/*metabolism/pathology ; Grifola ; HT29 Cells ; Humans ; Inflammatory Bowel Diseases/chemically induced/*drug therapy/pathology/physiopathology ; Interleukin-8/biosynthesis/genetics ; Intestinal Mucosa/*drug effects/metabolism/pathology ; Monocytes/*drug effects/metabolism/pathology ; NF-kappa B/genetics/metabolism ; Peroxidase/metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species/metabolism ; Stomach Ulcer ; Transcription, Genetic/drug effects ; Trinitrobenzenesulfonic Acid/administration & dosage ; Tumor Necrosis Factor-alpha/*biosynthesis/genetics ; U937 Cells ; Weight Loss

The size variation of the cytoxin-associated protein (cagA), which is dependent on the 3' repeat region (3'RR) of the cagA gene, is known to play a crucial role in the pathogenesis of Helicobacter pylori infection. The present study evaluated the relationship between the 3'RR variation and the geographic distribution, clinical manifestations, and locations of colonization in the stomach. We evaluated the 3'RR of H. pylori isolates from 78 patients with gastric cancer, peptic ulcer, and non-ulcer dyspepsia from Japan, Hong Kong, India, and the United States and assessed the variations of 3'RR according to the geographical and clinical characteristics. Sixty eight (87.2%) patients had the same 650 bp band without geographical differences. The frequency of polymorphisms in the 3'RR did not differ when compared to the clinical manifestations (P=0.868). The length of 3'RR did not differ by location of colonization. In conclusion, the 3'RR variation of cagA gene is not associated with the geographical and clinical characteristics of the patients studied.
Amino Acid Sequence ; Antigens, Bacterial/*genetics ; Bacterial Proteins/*genetics ; Dyspepsia/etiology ; Helicobacter Infections/diagnosis ; Helicobacter pylori/*genetics ; Humans ; Integration Host Factors ; Molecular Sequence Data ; Peptic Ulcer/etiology ; Polymorphism, Genetic ; Repetitive Sequences, Amino Acid ; Repetitive Sequences, Nucleic Acid ; Stomach Neoplasms/etiology

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

OBJECTIVEThis study was designed to investigate the interaction between pepsinogen C(PGC) insertion/deletion polymorphism and Helicobacter pylori(Hp) infection, together with its different subtype strains, in the development of gastric cancer (GC).

METHODSPGC Genotypes were determined by polymerase chain reaction (PCR) assay in 564 subjects with superficial gastritis (NOR), gastric ulcer (GU), atrophic gastritis (AG) and GC, who were frequency-matched as 1:1. Serum Hp-IgG antibodies were determined by an enzyme linked immunoadsorbent assay (ELISA). Hp genetic subtypes in 171 patients with Hp infection were determined by PCR methods.

RESULTSIn GU, AG and GC, the OR of interaction was 8.69 (P = 0.049), 11.16 (P = 0.02), and 10.61 (P = 0.03), respectively; the interaction index of PGC homozygous allele 1 genotype and Hp infection was 5.40, 6.48 or 4.34, respectively; the attributable proportions were 0.721, 0.770 and 0.697, respectively. In AG and GC, no significant interactions were observed between PGC polymorphism and Hp genetic subtypes.

CONCLUSIONThe findings of this study suggest that PGC insertion/deletion polymorphism and Hp infection seem to present a positive interaction in the development of gastric cancer. While no interactions may be present between PGC polymorphism and Hp genetic subtypes.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Base Sequence ; Female ; Gastritis ; genetics ; Gastritis, Atrophic ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Helicobacter Infections ; genetics ; Helicobacter pylori ; pathogenicity ; Humans ; INDEL Mutation ; Male ; Middle Aged ; Molecular Sequence Data ; Pepsinogen C ; genetics ; Polymorphism, Genetic ; Stomach Neoplasms ; genetics ; microbiology ; Stomach Ulcer ; genetics ; Young Adult

OBJECTIVETo observe the clinical effects of Jianwei Yuyang Granule (JYG) in treating patients with gastric ulcer and its influence on interleukin-1beta (IL-1beta) and basic fibroblast growth factor (bFGF) mRNA expression in gastric mucosa for exploring the therapeutic mechanism.

METHODSFifty-six patients with confirmed gastric ulcer unader gastroscope and differentiated as Gan-stagnant Pi-deficient syndrome were randomly assigned to two groups, the treated group (26 cases) treated with JYG and the control group (30 cases) treated with famotidine and sucralfate, 4 weeks as one therapeutic course. The changes before and after treatment in clinical compliance, symptom integral, ulcer-healing rate, clinical effective rate, and HP-clearance rate were observed. And the gastric mucosa biopsy was fetched for morphological examination and IL-1beta and bFGF mRNA expression detection by RT-PCR as well.

RESULTSThe clinical compliance rate in the treated group was 100 %, which was obviously better than that in the control group (86.7 %, P< 0.01); the improvement on symptom integral in the former was also better (P < 0.01); no statistical significance was shown in ulcer-healing rate, clinical effective rate and HP-clearance rate between the two groups. Morphological observation showed markedly decreased inflammatory cell infiltration, epithelial cell regeneration and rather regular glandular arrangement in both groups. The IL-1beta mRNA expression level decreased and that of bFGF increased in both groups after treatment significantly ( P < 0.01), but showed insignificant difference between the two groups.

CONCLUSIONJYG, with its good clinical compliance, has favorable effects in relieving clinical symptoms, promoting endoscopic ulcer healing and HP clearance, decreasing the expression of IL-1beta mRNA and increasing the expression of bFGF, therefore, it could promote the recovering of gastric ulcer.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fibroblast Growth Factor 2 ; genetics ; Gastric Mucosa ; drug effects ; metabolism ; pathology ; Humans ; Interleukin-1beta ; genetics ; Male ; Middle Aged ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Ulcer ; drug therapy ; Young Adult

OBJECTIVETo observe the effect of selective cyclooxygenase-2 (COX-2) inhibitor on the healing of experimental gastric ulcer in rats and explore its mechanisms in light of gastric acid secretion.

METHODSGastric ulcers were induced in rats by an application of acetic acid to the serosal surface of the anterior gastric body. The effects of selective COX-2 inhibitor, celecoxib, on the healing of gastric ulcer, the total acidity of gastric juice, the expressions of H+, K+-ATPase mRNA and protein, and the ultrastructure of the parietal cell were observed in comparison with the effects of normal saline.

RESULTSNine days after ulcer induction, the ulcer area was 11.9-/+3.1 mm square and 19.7-/+3.8 mm square in rats with normal saline and celecoxib treatments, respectively (P<0.01). The total acidity of gastric juice and the expressions of H+, K+-ATPase mRNA and protein in celecoxib group were significantly higher than that in normal saline group at both 6 and 9 days after ulcer induction, but no significant difference was found between the two groups in the amount of secretary canaliculus and microvillus.

CONCLUSIONSelective COX-2 inhibitor can significantly delay the healing of experimental gastric ulcer in rats, the mechanism of which might be associated with enhanced digestive action of gastric acid on the new granulation tissue at the ulcer base as a result of celecoxib-stimulated gastric acid secretion of the parietal cells.

Animals ; Celecoxib ; Cyclooxygenase 2 Inhibitors ; pharmacology ; therapeutic use ; Gastric Acid ; secretion ; Gene Expression Regulation, Enzymologic ; drug effects ; H(+)-K(+)-Exchanging ATPase ; genetics ; metabolism ; Hydrogen-Ion Concentration ; Male ; Microvilli ; drug effects ; pathology ; Parietal Cells, Gastric ; drug effects ; ultrastructure ; Pyrazoles ; pharmacology ; therapeutic use ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Stomach Ulcer ; drug therapy ; metabolism ; pathology ; Sulfonamides ; pharmacology ; therapeutic use