Ulcer Model Research Articles

Transcriptional landscape of intestinal environment in DSS-induced ulcerative colitis mouse model.

Ulcerative colitis (UC) is a chronic inflammatory disease that targets the colon and has seen an increasing prevalence worldwide. In our pursuit of new diagnostic and therapeutic approaches for UC, we undertook a sequencing of colons from UC mouse models. We focused on analyzing their differentially expressed genes (DEGs), enriching pathways, and constructing protein-protein interaction (PPI) and Competing Endogenous RNA (ceRNA) networks. Our analysis highlighted novel DEGs such as Tppp3, Saa3, Cemip, Pappa, and Nr1d1. These DEGs predominantly play roles in pathways like cytokine-mediated signaling, extracellular matrix organization, extracellular structure organization, and external encapsulating structure organization. This suggests that the UC pathogenesis is intricately linked to the interactions between immune and non-immune cells with the extracellular matrix (ECM). To corroborate our findings, we also verified certain DEGs through quantitative real-time PCR. Within the PPI network, nodes like Stat3, Il1b, Mmp3, and Lgals3 emerged as significant and were identified to be involved in the crucial cytokine-mediated signaling pathway, which is central to inflammation. Our ceRNA network analysis further brought to light the role of the Smad7 Long non-coding RNA (lncRNA). Key MicroRNA (miRNAs) in the ceRNA network were pinpointed as mmu-miR-17-5p, mmu-miR-93-5p, mmu-miR-20b-5p, mmu-miR-16-5p, and mmu-miR-106a-5p, while central mRNAs included Egln3, Plagl2, Sema7a, Arrdc3, and Stat3. These insights imply that ceRNA networks are influential in UC progression and could provide further clarity on its pathogenesis. In conclusion, this research deepens our understanding of UC pathogenesis and paves the way for potential new diagnostic and therapeutic methods. Nevertheless, to solidify our findings, additional experiments are essential to confirm the roles and molecular interplay of the identified DEGs in UC.

Curcumin regulates inflammation and apoptosis through PARP-1 and NF-κB in ethanol-induced gastric ulcer model

Curcumin regulates inflammation and apoptosis through PARP-1 and NF-κB in ethanol-induced gastric ulcer model

Investigation of the Protective Effects of Capparis Spinosa Extract in Indomethacin Induced Ulcer Model in Rats

ABSTRACT
 Objective: Capparis spinosa is a genus of the Capparaceae family. In the literature, it has been found that the main components of Capparis buds, quercetin and kaempferol were found to be effective in many diseases. In this study, the protective effects of Capparis spinosa on damaged rat stomach tissue induced by indomethacin and some antioxidant parameters were investigated.
 Material and Methods: A total of 36 female Sprague Dawley rats weighing 200-220 grams were used in the study and six groups were formed. Groups were: healthy group; positive control group (famotidine 20 mg/kg + indomethacin 25 mg/kg); negative control group (distilled water + indomethacin); low-dose study group (125 mg/kg Capparis spinosa + indomethacin); medium dose group (250 mg/kg Capparis spinosa + indomethacin); and high dose group (500 mg/kg Capparis spinosa + indomethacin). Six hours after indomethacin was given to the groups by gastric lavage, all rats were killed under general anesthesia. The stomachs of all rats were removed, the ulcerated areas on the stomach surface were evaluated macroscopically, and the ulcer areas were measured on mm2 paper. In addition, blood and stomach tissues of all rats were biochemically examined, and malondialdehyde, superoxide dismutase and glutathione parameters were measured. The antiulcer activity of CS was compared with all groups.
 Results: When the ulcer area and histopathological evaluation were examined, it was determined that the group applied 250 mg/kg Capparis spinosa had an appearance close to the healthy group. It was also found that plant extracts at all concentrations decreased the level of MDA in rat gastric tissue and increased SOD activity and GSH levels statistically.
 Conclusion: It was obtained that Capparis spinosa has antiulcer activity.

ENHANCING GASTRIC ULCER MANAGEMENT: NOVEL INSIGHTS FROM TERAZOSIN-PANTOPRAZOLE COMBINATION THERAPY

In this present study, we explored a novel approach to gastric ulcer management by investigating the therapeutic potential of terazosin, an alpha-1 adrenergic receptor inhibitor, in combination with pantoprazole, a common anti-ulcer agent. Employing an ethanol-induced rat-gastric ulcer model, the study demonstrated that terazosin pre-treatment significantly reduced ulcer formation, with the terazosinpantoprazole combination exhibiting superior mucosal protection compared to pantoprazole alone. Histopathological analysis revealed preserved mucosal structure and reduced neutrophil infiltration, indicating an anti-inflammatory effect. At a molecular level, the combination treatment groups exhibited elevated levels of phosphoglycerate kinase 1 (PGK-1), a vital enzyme in cellular energy metabolism, while inflammatory markers IκB kinase (IKK) and interleukin- 6 (IL-6) were significantly reduced, signifying mitigation of inflammation. These findings of the three different combinations of terazosin with pantoprazole indicate that this can be a potential approach for the treatment of gastric ulcers and can help in reducing the existing pantoprazole dose.

P131 Therapeutic effects of KPG-818 in a trinitrobenzene sulfonic acid - induced mouse Crohn's/Ulcerative Colitis model

Abstract Background Immunomodulatory drugs have the potential therapeutic benefit not only in malignancy but also in diseases characterized by an inflammatory phenotype. In inflammatory bowel diseases (IBD), colonic inflammation results from dysregulation of the mucosal immune responses to intestinal flora together with genetic and environmental factors. Inflammatory cytokines are implicated in pathogenesis of IBD. Treatments for IBD that improve clinical outcomes and limit adverse effects are needed. KPG-818 is a novel generation of lenalidomide derivative and small molecule immunomodulatory drug. KPG-818 is currently under clinical development for therapy of multiple myeloma, lymphoma (phase 1), and systemic lupus erythematosus (SLE) (phase 2a). Remarkable immunomodulatory effects of KPG-818 are observed in SLE patients. In preclinical studies, KPG-818 significantly inhibits inflammatory cytokines interleukin-6 and TNF-α in human peripheral blood mononuclear cells, inhibits tumor growth in multiple myeloma and lymphoma xenograft tumor models. Here we report the significant therapeutic effects of KPG-818 in a chronic Crohn’s/ulcerative colitis mouse model. Methods Trinitrobenzene Sulfonic Acid (TNBS) was intra-colonially administered to induce Balb/c mouse Crohn’s/ulcerative colitis model at 50 mg/kg, once a week for 4 weeks. KPG-818 was orally administered at doses of 0.03, 0.12, 0.4, and 1.0 mg/kg, once a day for 6 consecutive weeks. Upadacitinib served as a positive control drug. Vehicle solution served as a negative control. To evaluate the therapeutic effects of the KPG-818, disease activity index (DAI) of ulcerative colitis, the degree of weight loss, fecal morphology, and rectal bleeding were scored once every other day for six consecutive weeks. At the end of the study, colonic samples were collected for gross observation and scoring of macroscopic damage, measurement of length and weight, and histopathological diagnosis and scoring. At the end of study, peripheral blood samples were also collected for analyses of immune cells and cytokines. Results Compared to the vehicle control, KPG-818 significantly improved the body weight loss, disease activity of ulcerative colitis, gross degree of colorectal injury, pathological lesions of colorectal tissue, inhibited the inflammatory cytokines in a dose-response manner. The effective dose was as low as 0.03 mg/kg. KPG-818 showed similar therapeutic effects to Upadacitinib. Conclusion In this mouse model of chronic Crohn’s/ulcerative colitis, KPG-818 showed remarkable therapeutic effects in a dose-dependent manner. These data encourage the clinical evaluation of KPG-818 in inflammatory bowel diseases.

S100a10 deficiency in neutrophils aggravates ulcerative colitis in mice

Background and aimsS100a10 is a member of the S100 family of proteins, which plays a key role in the depression and tumor metastasis. However, the role of S100a10 is unclear in ulcerative colitis. MethodsThe effect of S100a10 was assessed using a murine ulcerative colitis model which was accompanied by parameters including body weight loss, disease activity index, histological score, colon weight and length. The quantity and role of immune cells was determined by flow cytometry and bone marrow chimeric mice. Neutrophils depletion, adoptive cell transfer and conditional knockout mice were used to ascertain which cells played the key role in ulcerative colitis. The function of neutrophils was evaluated by migration assay, phagocytosis assay, multiplex immunoassay and real-time PCR. ResultsIn this study, our data showed that S100a10-/- mice were prone to ulcerative colitis induced by dextran sodium sulfate. Neutrophils number increased in colon of S100a10-/- mice after dextran sodium sulfate treatment significantly. Meanwhile, adoptive transfer of neutrophils from wild type mice partially decreased the susceptibility of S100a10-/- mice to dextran sodium sulfate. There was no difference in ulcerative colitis between the groups of S100a10-/- mice without neutrophils and wild type mice. Finally, we found that S100a10-/- neutrophils had stronger function in secretion and synthesis of inflammatory factor. ConclusionsIn one word, these results suggest that S100a10 has a role in inhibiting the pathogenesis of ulcerative colitis through regulation of neutrophils function.

Experimental animal models for gastric ulcer / peptic ulcer: An overview

In the present study we have discussed around sixteen different animal models used worldwide for the scientific research and new drug discovery. The main aim of the using experimental animal models in drug discovery is to establish and provide evidence for non-clinical 'proof-of-concept' for the safety, efficacy, and target of interest for specific drug molecules. Experimental preparations developed in one species for the purpose of studying phenomena occurring in another species. The use of experimental animal models serves to better understand the origins, pathology, and the overall nature of comparable diseases of humans being. Similarly, animal models perform duties for in the development of safe and effective treatments and cures of such diseases and/or associated symptoms. Experimental animal models for drug discovery and development have played a major role in the characterization of the pathophysiology of diseases and associated mechanisms of injury, drug target identification, and evaluation of novel therapeutic agents for toxicity, pharmacokinetics and pharmacodynamics activity. Through animal model researchers can perform experiments that would be impractical or ethically prohibited with humans. There are various animal models used for screening of uncountable therapeutic activities, in this review our main focus is animal models used for peptic ulcer. Peptic ulcer is one of the worldwide diseases where 10% of adults are affected by peptic ulcer once in their lifetime. The antiulcer models for drug development against gastric and duodenal ulcer studies are limited in number that has hindered the progress of targeted therapy in this field. Therefore, it is necessary to review the literature on experimental animal models that are used to screen agents with potential anti-gastric ulcer activity and describe their biochemical basis in order to facilitate their use in the development of new preventive and curative antiulcer drugs. There are many models used to induce ulcer such as pylorus ligation or it can be chemically induced by ethanol, NSAIDs (e.g. indomethacin) or many more. In this review paper, current in-vivo animal models of ulcers and the pathophysiological mechanisms underlying their induction, their drawbacks, as well as the challenges associated with their use have been discussed.

Rumex nervosus nanoparticles versus aqueous extract: introduction of novel immunomodulatory anti-inflammatory nano-preparation for management of paw oedema and gastritis in rats

BackgroundRumex nervosus is abundant in East African and Arabian countries, and is used in curing gastrointestinal diseases as well as in wound healing. The current study intends to assess Rumex nervosus aqueous extract's anti-inflammatory benefits compared to its nanoparticles on rat paw edema and its gastric mucosal protection against ulceration.Materials and methodsIn-vitro cytotoxicity effects and antioxidant activity of Rumex nervosus nanoparticles versus aqueous extract were studied, followed by a pilot in vivo pharmacological study to determine the suitable dose of nanoparticles that would be used in the safety and efficacy studies in comparison with the aqueous extract. Its protective effects on arthritis and soft tissue inflammation were studied in rat paw edema and gastric ulcer models. Rumex nervosus extract (250 and 500 mg/kg) and nanoparticles (3.3 and 6.6 mg/kg) were given to four groups of rats orally before induction of paw oedema with subplantar 0.2 ml (1% w/v) formaldehyde or gastritis with oral ethanol 1 ml (70%), besides negative, positive control and reference groups.ResultsPaw volumes and gastric ulcer indices, as well as the anti-inflammatory and antioxidant parameters (kappa β, Paraoxonase1, and Malondialdehyde) that were measured in sera showed a marked reduction in groups treated with high doses of Rumex nervosus extract, and nanoparticles. Histopathologic and histochemical assessment of the stomachs confirmed the other investigations. All results were significant compared to positive control untreated groups.ConclusionsMost studies demonstrated Rumex nervosus’s protective anti-inflammatory benefits with the superiority of large doses of nanoparticles, offering a promising natural solution for low cost against inflammation.

C-Reactive Protein Is Not the Driver Factor in Ulcerative Colitis.

Purpose: C-reactive protein (CRP) functions as a nonspecific marker in various inflammatory disorders, particularly in evaluating the efficacy of pharmacological treatments in patients with ulcerative colitis. The existing body of evidence does not offer adequate support for the direct implication of CRP in modulating the advancement of ulcerative colitis. Methods: Our study employed a rigorous mouse model. An ulcerative colitis mouse model was established by subjecting CRP-deficient mice to dextran sulfate sodium (DSS) treatment. The phenotype of the mice, which encompassed parameters such as body weight, colon length, and spleen weight, was meticulously evaluated. Additionally, various physiological and biochemical indicators were assessed, including colon histopathology, expression levels of inflammatory factors, and staining of the intestinal mucus layer. Results: The absence of CRP did not significantly affect the phenotype, physiological characteristics, and biochemical indices in a mouse model of ulcerative colitis compared to mice with wild-type CRP. Additionally, eliminating intestinal bacteria flora interference through antibiotic treatment revealed that mice lacking CRP did not demonstrate any notable variations in the ulcerative colitis model. Meanwhile, the survival rate of mice lacking CRP did not exhibit a statistically significant difference compared to wild-type mice. Conclusion: The results of our study suggest that CRP may not directly mediate ulcerative colitis. Instead, it is more likely to be a bystander that is present alongside with elevated inflammatory factors. Further investigation is warranted to determine the precise role of CRP in humans, given the significant limitations associated with the use of mouse models.

Histopathological and biochemical evaluation of the protective efficacy of Prunus spinosa L. extract in a rat model of indomethacin-induced gastric ulcer.

Some species of Prunus L. are popularly used to treat gastric ulcers. However, the possible healing mechanisms of the anti-ulcer activity of P. spinosa, which has proven antioxidant, anti-inflammatory, and wound-healing properties, are unclear. Ethanol extracts of P. spinosa fruits were administered orally at 100 mg/kg and 200 mg/kg to Wistar albino rats, with an indomethacin-induced gastric ulcer model. The ulcerous areas on the stomach surface were examined macroscopically. Tissues were examined histopathologically and biochemically. LC-HRMS revealed the phytochemical content. TNF-α, IL-6, IL-1β, IL-8, and NF-kB levels were higher in the gastric ulcer group than in the extract groups. The VEGF values did not differ in each group. A significant difference was found between the lansoprazole group and the high-dose P. spinosa group regarding PGE2 levels. A histopathologically significant difference was observed between the healthy group and the indomethacin-applied groups in terms of neutrophilic infiltration of the gastric mucosa. Ascorbic acid (1547.521 µg/g), homoprotocatechuic acid (1268.217 µg/g), and genistein (1014.462 µg/g) were found as the main compounds in the P. spinosa extract by LC-HRMS. Our results demonstrated that P. spinosa protected the gastric mucosa from inflammation and also modulated the PGE2 pathway. When considered in terms of TNF-α, IL-1β, IL-8, IL-6, PGE2, and NF-kB values, it can be concluded that it has a similar or even more positive effect than the reference substance. P. spinosa showed its effects in a dose-dependent manner.

Metabolomics of interventional effects of Coptidis Rhizoma and its processed products on oral ulcer due to excess heat in rats

Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q/TOF-MS) was employed to examine the impact of Coptidis Rhizoma(CR) and its processed products on the metabolism in the rat model of oral ulcer due to excess heat and to compare the effectiveness of CR and its three products. Male SD rats were randomly allocated to the sham-operation(Sham), model(M, oral ulcer due to excess heat), CR, wine/Zingiberis Rhizoma Recens/Euodiae Fructus processed CR(wCR/zCR/eCR), and Huanglian Shangqing Tablets(HST) groups. Except the Sham group, the other groups were administrated with Codonopsis Radix-Astragali Radix decoction by gavage for two consecutive weeks. The anal temperature and water consumption of rats were monitored throughout the modeling period of excess heat. Following the completion of the modeling, oral ulcer was modeled with acetic acid. Hematoxylin-eosin(HE) staining was employed to observe the mucosal pathological changes in oral ulcer. A colorimetric assay was employed to determine the serum level of glutathione peroxidase(GSH-Px). Enzyme-linked immunosorbent assay(ELISA) was conducted to determine the levels of tumor necrosis factor-alpha(TNF-α), interleukin-6(IL-6), interleukin-1β(IL-1β), superoxide dismutase(SOD), and malondialdehyde(MDA) in the serum. The non-targeted metabolomics analysis based on UPLC-Q/TOF-MS was conducted on the serum samples. Metabolic profiles were then built, and the potential biomarkers were screened by principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). The Mev software was used to establish a heat map and conduct cluster analysis on the quantitative results of the markers. The online databases including MBRole, KEGG, and MetaboAnalyst were used for pathway enrichment analysis and metabolic network building. The experimental results showed that the modeling led to pathological damage to the oral mucosa, elevated serum levels of TNF-α, IL-6, IL-1β, and MDA, and lowered levels of SOD and GSH-Px in rats. The drug administration recovered all the indices to varying extents, and wCR exhibited the best performance. Non-targeted metabolomics identified 48 differential metabolites including 27 metabolites in the positive ion mode and 21 metabolites in the negative ion mode. Five enriched pathways were common, including glycerophospholipid metabolism, linoleic acid metabolism, and tyrosine metabolism. Conclusively, CR and its three processed products could alleviate the inflammation and oxidative stress injury in rats suffering from oral ulcers due to excess heat by regulating lipid and amino acid metabolism. Notably, wCR demonstrated the most significant therapeutic effect.

Lactiplantibacillus plantarum HY7718 Improves Intestinal Integrity in a DSS-Induced Ulcerative Colitis Mouse Model by Suppressing Inflammation through Modulation of the Gut Microbiota.

Inflammatory bowel disease (IBD), a chronic condition that causes persistent inflammation in the digestive system, is closely associated with the intestinal microbiome. Here, we evaluated the effects of Lactiplantibacillus plantarum HY7718 (HY7718) on IBD symptoms in mice with dextran sulfate sodium (DSS)-induced colitis. Oral administration of HY7718 led to significant improvement in the disease activity index score and the histological index, as well as preventing weight loss, in model mice. HY7718 upregulated the expression of intestinal tight junction (TJ)-related genes and downregulated the expression of genes encoding pro-inflammatory cytokines and genes involved in the TLR/MyD88/NF-κB signaling pathway. Additionally, HY7718 reduced the blood levels of pro-inflammatory cytokines, as well as reversing DSS-induced changes to the composition of the intestinal microbiome. HY7718 also increased the percentage of beneficial bacteria (Lactiplantibacillus and Bifidobacterium), which correlated positively with the expression of intestinal TJ-related genes. Finally, HY7718 decreased the population of pathogens such as Escherichia, which correlated with IBD symptoms. The data suggest that HY7718 improves intestinal integrity in colitis model mice by regulating the expression of TJ proteins and inflammatory cytokines, as well as the composition of the intestinal microflora. Thus, L. plantarum HY7718 may be suitable as a functional supplement that improves IBD symptoms and gut health.

Wuwei Kushen Changrong capsule alleviates DSS-induced colitis in mice via inhibition of NLRP3 inflammasome and STAT3 pathway.

Wuwei Kushen Changrong capsule (Composite Sophora Colon-soluble Capsule, CSCC) is a Chinese patent medicine developed to treat ulcerative colitis. Studies highlight CSCC potential efficacy for ulcerative colitis (UC) but unclear mechanism limits its widely treatment for patients. We aimed to investigate the anti-colitis efficacy of CSCC and explore the mechanism by which GPR43 inhibits the NLRP3/STAT3 signaling pathway, thereby mediating the protective effects of CSCC on the intestinal barrier. The protective effects of CSCC were evaluated in a murine ulcerative colitis model induced by 3% DSS. Assessments included body weight, Disease Activity Index (DAI) score, colon length, and histopathological score. Colon tissue, cell function, and immune-inflammatory status were evaluated using immunohistochemistry, immunofluorescence, ELISA, and real-time fluorescence quantitative PCR (RT-PCR). Protein expression levels of relevant pathways and receptors were measured using Western blot. All experiments were repeated. CSCC protected mice from DSS-induced colitis by upregulating Gpr43, promoting the expression of ZO-1 and Occludin tight junction proteins. Mechanistically, CSCC inhibits the MEK4/JNK1/STAT3 activation pathway, consequently suppressing the STAT3/NLRP3/IL-1β pathway and inhibiting the production of inflammatory factors such as IL-17A. The mechanisms through which CSCC protects against DSS-induced colitis may include upregulating Gpr43, inhibiting the STAT3/NLRP3 pathway, and suppressing inflammation factors like IL-17A. These findings highlight the mechanisms underlying CSCC's anti-colitis effects and suggest its potential as a therapeutic candidate for managing the progression of UC.

Screening of anti-ulcer activity of Emblica officinalis fruit extract on NSAID-induced peptic ulcer model in albino rats

Background: Peptic ulcer disease (PUD) is a prevalent health concern. Although proton pump inhibitors (PPIs), H2 receptor blockers, prostaglandin analogs, etc., are available as anti-ulcer agents, they have their own limitations. The clinical assessment of these medications revealed adverse effects, relapse frequency, and tolerance development, raising doubts about their effectiveness. Thus, there is a need to search for newer synthetic and herbal formulations that have fewer side effects and improved efficacy. Emblica officinalis (EO) or amla has shown to have antioxidant and antisecretory properties; hence, it was chosen for the study. Aims and Objectives: (1) The aim of this study was to screen the anti-ulcer activity of EO aqueous fruit extract in non-steroidal anti-inflammatory drug-induced peptic ulcer models of albino rats and (2) to compare the anti-ulcer effect of EO aqueous fruit extract with the standard drug omeprazole. Materials and Methods: Four groups comprising six albino rats each were formed. Group 1 (the control) received distilled water, followed by indomethacin 25 mg/kg orally. Group 2 (standard) received omeprazole 10 mg/kg, followed by indomethacin 25 mg/kg. Groups 3 and 4 received the aqueous extract of EO in 250 mg/kg and 500 mg/kg doses, respectively, followed by indomethacin at 25 mg/kg. After 1 h of indomethacin administration, the stomach was examined for ulceration. The severity score was determined to calculate the ulcer index (UI) and percentage inhibition. The results were analyzed statistically using the Mann–Whitney U-test (SPSS software version 22). Results: In the screening model mentioned above, the test agent at dosages of 250 mg/kg and 500 mg/kg body weight resulted in a significant decrease in total severity score, UI, and percentage inhibition as compared to the control group. The comparison of the results of the standard drug omeprazole with those of the test groups was statistically insignificant. Conclusion: The gastroprotective activity of amla (EO) may be due to its antioxidant and antisecretory properties. This study proves the gastroprotective action of EO.

Evaluation of Synergistic Activity of Ethanolic Leaf Extracts of Tephrosia Purpurea and Bacopa Monnieri in Ulcer Induced Rats.

To evaluate synergistic antiulcer activity of ethanolic extracts of Tephrosia purpurea and Bacopa monnieri in ulcer induced rats. Ethanolic leaf extracts of both the plants were administered individually and in combination at a dose of 200mg/kg to ulcer induced male albino rats. Omeprazole 10mg/kg was used as standard. Pylorus ligation method, ethanol and indomethacin induced gastric ulcer models were the different gastric ulcer models selected for the induction of ulcer in rats. Ulcer index, ulcer score, total acidity, pH, percentage protection, volume of gastric juice were the parameters evaluated and compared in different groups in all the models. Decrease in the ulcer score, ulcer index, total acidity was observed and percentage protection was significant(*p<0.05 and p<0.01) with the combination extract compared to group received individual plant extracts. Our results suggested that combination of two medicinal plants showed synergistic anti ulcer activity and decreased the formation of ulcer lesions in rats.

Targeting NF-κB/COX-2 signaling by soyasaponin I alleviates diclofenac-induced gastric ulceration in male albino rats.

Gastric ulceration is a prevalent worldwide clinical presentation due to altered gastric defense mechanisms. Nonsteroidal anti-inflammatory drugs are one of the common causes of gastric ulcers mediated by the release of inflammatory mediators. The study aimed to investigate the potential protective effect of soyasaponin I (soya) against diclofenac (DIC)-induced gastric ulcer in rats and to highlight the underlying mechanisms. The experiment was conducted on 40 male Wistar albino rats, equally distributed into five groups: control, DIC-induced ulcer (9 mg/kg/d, orally, twice daily for 3 days), ulcer/soya-, ulcer/ranitidine-, and ulcer/soya/selective nuclear factor kappa B inhibitor (JSH-23)-treated groups. The doses of soya, ranitidine, and JSH were 20, 25, and 5 mg/kg/d, respectively, given orally. Gastric specimens were prepared for gene and histological study and for biochemical analysis of gastric prostaglandin E2 (PGE2), oxidative markers, and inflammatory cytokines. The gastric samples were formalin-fixed, paraffin-embedded, and subjected to hematoxylin and eosin (H&E), PAS staining, and immunohistochemical assay for identification of nuclear factor kappa B (NF-κB), cyclooxygenase-2 (COX-2), and proliferation marker (Ki67) expressions. The findings revealed decreased gastric PGE2 and altered inflammatory and oxidative markers in the ulcer model group. The H&E staining showed mucosal injury characterized by mucosal surface defects and inflammatory cell infiltrations. The polymerase chain reaction (PCR) and immunohistochemistry demonstrated an upregulation of NF-κB and COX-2 expression at gene/protein levels; meanwhile, Ki67 downregulation. The soya-treated group showed maintained biochemical, histological, and PCR findings comparable to the ranitidine-treated group. The JSH-23-treated group still showed partial gastric protection with biochemical and immunohistochemical changes. Soyasaponin I ameliorated DIC-induced gastric ulcers by targeting the COX-2 activity through modulation of NF-κB signaling.

Antiulcer activity of ethanolic extract of Arctocarpus hirsutus Lam. leaves in albino Wistar rats

Peptic ulcers, a prevalent gastrointestinal disorder, remain a significant health concern. Artocarpus hirsutus Lam., a plant deeply rooted in traditional medicine, has been historically employed to address various health issues. Each component of this plant showcases diverse therapeutic activities, offering a holistic approach to health concerns. This study aimed to explore the gastroprotective and antiulcerogenic potential of the ethanol-based extract derived from Artocarpus hirsutus Lam leaves. Administered orally in 100, 200, and 400mg/kg doses, the extract's effects were compared with established medications - omeprazole (30mg/kg) and sucralfate (100mg/kg). Parameters such as gastric acid volume, gastric pH, ulcer index, total acidity, and free acidity were evaluated. The ethanol-based extract derived from Artocarpus hirsutus Lam. leaves demonstrated activity at doses of 200mg/kg and 400mg/kg, resulting in a marked decrease in free acidity, total acidity, gastric volume, ulcer index, and an increase in gastric pH compared to the ulcer control group. Notably, the ethanol-based extract of leaves of Artocarpus hirsutus Lam. exhibited significant gastroprotective and ant-ulcerogenic effects in both ethanol induced ulcer and pylorus ligated ulcer models. These findings underscore the potential therapeutic value of Artocarpus hirsutus Lam. as an effective agent against peptic ulcers, supporting its traditional use in holistic health practices.

The Effect of Washing After Protein Blocking on Immunohistochemical Staining Applied to Paraffin Sections

Aim: Immunohistochemistry is still frequently preferred in both diagnostic and experimental studies&#x0D; because it can show proteins where they are in the tissue. One of the main problems in immunohistochemistry&#x0D; is the background staining, which can be prevented by protein blocking and which occurs as&#x0D; a result of binding of primary antibodies to tissue proteins and Fc receptors in the tissue due to antigenic&#x0D; similarity. There is no consensus on whether to wash after protein blocking in current publications and&#x0D; immunohistochemistry manuals published by manufacturers.&#x0D; Material and Methods: In our study, routine immunohistochemistry procedure was applied to determine&#x0D; the expression of TNF-α on 5 μm thick sections obtained from rat gastric tissue samples in which an&#x0D; experimental gastric ulcer model was created with ethanol, and two groups were formed, with and without&#x0D; washing after protein blocking, with 10 sections in each group. For semi-quantitative evaluation, the&#x0D; histological score (h-score) was calculated from the images obtained from the immunohistochemically&#x0D; stained preparations of both groups and the obtained data were statistically compared.&#x0D; Results: As a result of our study, no statistically significant difference was found between the h-score&#x0D; values of the groups that were washed after protein blocking in immunohistochemical staining and&#x0D; the groups that did not wash (p=0.971). The median (min-max) values of the groups with and without&#x0D; washing are 211 (179-244) and 215 (171-251), respectively.&#x0D; Conclusion: In the immunohistochemical staining procedure, washing after protein blocking does not&#x0D; change the staining intensity, does not create background staining, and does not affect the result of&#x0D; the h-score for semiquantitative evaluation. Similar studies are recommended for other species, tissue&#x0D; types and antigens.

Hippocampal and gut AMPK activation attenuates enterocolitis-like symptoms and co-occurring depressive-like behavior in ulcerative colitis model mice: Involvement of brain-gut autophagy

Patients with inflammatory bowel disease, including ulcerative colitis (UC) and Crohn's disease, have a high incidence of psychiatric disorders, including depression and anxiety. However, the underlying pathogenic mechanism remains unknown. Dextran sulfate sodium (DSS)-treated mice, a model of UC, exhibit depressive-like behavior and reduced adenosine monophosphate-activated protein kinase (AMPK) activity, which regulates various physiological functions in the brain and gut. However, comprehensive studies on UC pathophysiology with co-occurring depression focused on brain-gut AMPK activity are lacking. Therefore, we aimed to investigate whether resveratrol (RES), an AMPK activator, prevented DSS-induced UC-like symptoms and depressive-like behavior. DSS treatment induced UC-like pathology and depressive-like behavior, as assessed via the tail suspension test. Moreover, western blotting and immunohistochemical studies revealed that DSS increased p-p70S6 kinase (Thr389), p62, tumor necrosis factor-α, interleukin (IL)-1β, IL-18, NLR family pyrin domain containing 3 (NLRP3), cleaved caspase-1, cleaved Gasdermin-D (GSDMD), and cleaved caspase-3 expression levels in the rectum and hippocampus, and increased CD40, iNOS, and Kelch-like ECH-associated protein 1 expression levels, and the number of Iba1-positive cells in the hippocampus, and decreased p-AMPK and LC3II/I expression levels, and the number of NF-E2-related factor 2 (Nrf2)-positive cells, and reduced neurogenesis in the hippocampus. These changes were reversed by the RES administration. RES also enhanced PGC1α and SOD1 expression in the hippocampus of DSS-treated male mice. Moreover, NLRP3 staining was observed in the neurons and microglia, and cleaved GSDMD staining in neurons in the hippocampus of DSS-treated mice. Notably, RES prevented UC-like pathology and depressive-like behavior and enhancement of autophagy, decreased rectal and hippocampal inflammatory cytokines and inflammasome, and induced the Nrf2-PGC1α-SOD1 pathway in the hippocampus, resulting in neurogenesis in the hippocampal dentate gyrus. Our findings suggest that brain-gut AMPK activation may be an important therapeutic strategy in patients with UC and depression.

Antiulcer activity of the stem bark aqueous extract of Croton oligandrum (Euphorbiaceae) against Ethanol/HCl-induced gastric mucosal injury in rats

Gastric ulcer is an alteration of the lining of the stomach with destruction of mucus-producing cells. Many factors are contributing to the development of peptic ulcer such as phygiological stress, high production of acid, non-steroidal anti-inflammatory drugs (NSAIDs) and alcohol consumption. Croton oligandrum is traditionally used to treat many diseases including gastric ulcers. This study aims to investigate the cytoprotective activity of Croton oligandrum. To assess the gastroprotective potential of Croton oligandrum ulcer models were established using ethanol and indomethacin. Sucralfate was used as reference anti-ulcer drugs. Rats were treated with Croton oligandrum before induction of gastric ulceration by ethanol/HCl or HCl-ethanol-Indomethacin. The area and ulcer index appearance of ulcers were quantified, and mucus was measured. Pretreatment with Croton oligandrum showed a remarkable decrease in ulcer area when compared with control groups. Croton oligandrum (50, 100 mg/kg and 200 mg/kg of body weight) exhibited gastroprotective activity and prevented both gastric lesions induced in rats.