Gastric Mucosal Cell Proliferation Research Articles

Research on the signaling pathway and the related mechanism of traditional Chinese medicine intervention in chronic gastritis of the "inflammation-cancer transformation".

Objective: The aim of this study is to uncover the traditional Chinese medicine (TCM) treatments for chronic gastritis and their potential targets and pathways involved in the "inflammation-cancer" conversion in four stages. These findings can provide further support for future research into TCM and its active components. Materials and methods: The literature search encompassed PubMed, Web of Science, Google Scholar, CNKI, WanFang, and VIP, employing keywords such as "chronic gastritis", "gastric cancer", "traditional Chinese medicine", "medicinal herb", "Chinese herb", and "natural plant". Results: Herbal remedies may regulate the signaling pathways linked to the advancement of chronic gastritis. Under the multi-target and multi-pathway independent or combined reaction, the inflammatory microenvironment may be enhanced, leading to repair of damaged gastric mucosal cells, buffering the progress of mucosal atrophic degeneration via the decrease of inflammatory factor expression, inhibition of oxidative stress-induced damage, facilitation of microvascular neovascularization in the gastric mucosa and regulation of the processes of gastric mucosal cell differentiation and proliferation. Simultaneously, the decreased expression of inflammatory factors may impact the expression of associated oncogenes and regulate the malignant proliferation of cells, thereby achieving the treatment and prevention objectives of gastric cancer through the reduction of cell metastasis and apoptosis. Conclusion: Chinese medicine formulations and individual drugs can be utilised at various stages of the "inflammation-cancer" progression of chronic gastritis to prevent and treat gastric cancer in a multi-level, multi-targeted, and multi-directional fashion. This can provide guidance for the accurate application of medicines during different stages of "inflammation-cancer" transformation. New insights into the mechanism of inflammation-cancer transformation and the development of novel drugs for chronic gastritis can be gained through an extensive investigation of TCM treatment in this condition.

Knocking Down HN1 Blocks Helicobacter pylori-Induced Malignant Phenotypes in Gastric Mucosal Cells and Inhibits Gastric Cancer Cell Proliferation, Cytoskeleton Remodeling, and Migration.

Helicobacter pylori (H. pylori) is implicated in the aberrant proliferation and malignant transformation of gastric mucosal cells, heightening the risk of gastric cancer (GC). HN1 is involved in the development of various tumors. However, precise mechanistic underpinnings of HN1 promoting GC progression in H. pylori remain elusive. The study collected 79 tissue samples of GC patients, including 47 with H. pylori-positive GC and 32 H. pylori-negative controls. Using human gastric epithelial cells (GES-1) and human gastric adenocarcinoma cells (HGC-27), the effect of overexpression / knocking down of HN1 and H. pylori infection was evaluated on cell function (proliferation, migration, apoptosis), cytoskeleton, and expression of cell malignant phenotype factors that promote the malignant biological behavior of cancer cells. The expression of HN1 in GC tissues is higher than that in paracancerous tissue and is closely related to infiltration, lymphatic metastasis, distant metastasis, survival, and H. pylori infection. Downregulation of HN1 effectively hinders the ability of H. pylori strains 26695 and SS1 to promote migration of GES-1 and HGC-27 cells, while lowering the expression of key indicators associated with malignant phenotype. Downregulated GSK3B, β-catenin, and Vimentin after knockdown Integrinβ1, but HN1 expression remained largely unchanged, when HN1 and Integrinβ1 were knocked down, GSK3B, β-catenin, and Vimentin expression were considerably reduced. Our research demonstrated the crucial role of HN1 in H. pylori-induced acquisition of a malignant phenotype in GES-1 cells. Knockdown of HN1 blocked the pathogenic mechanism of H. pylori-induced GC and downregulated the expression of GSK3Β, β-catenin and Vimentin via Integrin β1.

FTO-mediated m6A modification promotes malignant transformation of gastric mucosal epithelial cells in chronic Cag A+ Helicobacter pylori infection.

Cag A+ Helicobacter pylori chronic infection cause malignant transformation of the human gastric mucosa. N6-methyladenosine (m6A) modifications are the most common and abundant mRNA modifications and one of the pathways affecting tumorigenicity and tumor progression. However, the role of m6A modification in the process of chronic H. pylori infection leading to malignant transformation of gastric mucosa is unclear. In this study, we used Cag A- and Cag A+H. pylori chronic infection to establish cellular models in GES-1 cells and analyzed the cellular morphology, proliferation, apoptosis, invasiveness and tumorigenicity of gastric mucosal epithelial cells. The m6A expression levels of GES-1 cells after chronic infection with Cag A- and Cag A+H. pylori were examined, and modifying effect of FTO (the fat mass and obesity-associated protein) on CD44 was verified by MeRIP-qPCR. Finally, the FTO expression changes and m6A expression levels were further validated in clinical gastric cancer tissues. Chronic Cag A+H. pylori-infected GES-1 cells exhibit altered cell morphology, apoptosis inhibition, abnormal proliferation, enhanced migration, colony formation, and increased stem cell-like properties. Meanwhile, FTO and CD44 expression was enhanced, and FTO may induce malignant transformation of gastric mucosa by regulating CD44 mRNA m6A methylation modifications. We verified the effect of chronic stimulation of Cag A+H. pylori on malignant transformation of gastric mucosal epithelium. revealing the possibility of FTO in promoting malignant transformation of gastric mucosa by modifying CD44 mRNA methylation, suggesting that FTO expression is a potential molecule for malignant transformation of gastric mucosal epithelial cells.

The protective effect of NF-κB signaling pathway inhibitor PDTC on mice with chronic atrophic gastritis

Objective To understand the protective effect of NF-κB signaling pathway inhibitor pyrrolidinedithiocarbamate (PDTC) on mice with chronic atrophic gastritis (CAG). Methods Helicobacter pylori (H. pylori) infection combined with high-salt diet was used to construct the CAG mouse model, and 100 or 200 mg/kg/day PDTC was intragastrically treated for 8 weeks. Then, hematoxylin and eosin (HE) and Alcian blue-periodic acid-Schiff (AB-PAS) staining were used to observe the pathology of gastric mucosa, while immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), enzyme-linked immuno sorbent assay (ELISA) and western blotting were determined to detect the expression of related molecules. Results The nuclear content of NF-κB p65 in the gastric mucosa of the CAG mice was increased accompanying by the structural disorder of the gastric mucosal epithelium, inflammatory cell infiltration, intestinal metaplasia, and increased MUC2 expression, but the symptoms were alleviated after PDTC treatment. In addition, the expressions of TNF-α, IL-1β, IL-6 and COX2 in the gastric mucosa and serum of CAG mice were higher than those control mice, which were reduced in CAG mice treated with either 100 or 200 mg/kg PDTC. Furthermore, 100 mg/kg and 200 mg/kg PDTC treatments reduced the serum PGE2 in CAG mice with the decreased PCNA and Ki-67 expression in gastric mucosa. The therapeutic effect of 200 mg/kg PDTC was significantly better than that of 100 mg/kg PDTC. Conclusion PDTC inhibited inflammation and the excessive proliferation of gastric mucosal epithelial cells, thereby exerting a potential therapeutic effect on CAG.

Показатели пролиферации и апоптоза эпителиальных клеток слизистой оболочки желудка у коренных жителей Республики Тыва с Helicobacter pylori-позитивной язвенной болезнью двенадцатиперстной кишки

Показатели пролиферации и апоптоза эпителиальных клеток слизистой оболочки желудка у коренных жителей Республики Тыва с Helicobacter pylori-позитивной язвенной болезнью двенадцатиперстной кишки

Chronic CagA-positive Helicobacter pylori infection with MNNG stimulation synergistically induces mesenchymal and cancer stem cell-like properties in gastric mucosal epithelial cells.

A CagA-positive Helicobacter pylori (H. pylori) infection can cause malignant transformation of human gastric mucosal epithelial cells, and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) is a chemical carcinogen that induces gastric carcinogenesis. Whether this environmental chemocarcinogen may synergistically enhance the risk of H. pylori-infected gastric cancer remains unclear. In this study, we adopted a chronic CagA-positive H. pylori infection with or without MNNG coinduction to establish a cellular model in GES-1 cells and an animal model in C57BL/6J mice. The proliferation, cell phenotype, apoptosis, epithelial-mesenchymal transition (EMT), stemness and tumorigenicity of gastric mucosal epithelial cells were analyzed in vitro and in vivo. The results showed that chronic H. pylori-infected GES-1 cells displayed inhibited apoptosis, abnormal proliferation, enhanced invasion, and migration, increased EMT/mesenchymal phenotype, colony formation and stem cell-like properties, and enhanced tumorsphere-formatting efficiency as well as CD44 expression, a known gastric cancer stem cell (CSC) marker. MNNG synergistically promoted the above actions of chronic H. pylori infection. Further studies in chronic H. pylori-infected C57BL/6J mice models showed that an increased incidence of premalignant lesions in the gastric mucosa tissue of the H. pylori-infected mice had occurred, the mouse gastric mucosa cells exhibited similar mesenchymal and CSC-like properties in the above GES-1 cells, and precancerous lesions and EMT/CSC-like phenotypes were reinforced by the synergistic action of MNNG stimulation. H. pylori infection and/or MNNG induction were capable of causing enhanced expression and activation of Wnt2 and β-catenin, indicating that the Wnt/β-catenin pathway is involved in the actions of H. pylori and MNNG. Taken together, these findings suggest that chronic CagA-positive H. pylori infection with MNNG stimulation synergistically induces mesenchymal and CSC-like properties of gastric mucosal epithelial cells.

Effects of miR-101 on the proliferation and apoptosis of gastric mucosal epithelial cells via Nrf2/ARE signaling pathway.

The aim of this study was to investigate the effects of micro-ribonucleic acid (miR)-101 on the proliferation and apoptosis of gastric mucosal epithelial cells through nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway. Human gastric epithelial AGS (CRL-1739) cells were cultured in vitro. MiR-101 down-regulation or over-expression was achieved by transfection of inhibitors, or miRNA mimics, respectively. The apoptosis rate was detected by flow cytometry. Meanwhile, the targets of miR-101 were verified by the Dual-Luciferase reporter gene assay. Furthermore, the changes in protein levels were measured via Western blotting (WB). Up-regulation of miR-101 significantly promoted the apoptosis of gastric mucosal epithelial cells. The 3'-untranslated region (3'-UTR) of Nrf2 was highly conserved to combine with miR-101. The Luciferase reporter gene assay showed that transfection of miR-101 mimics could remarkably inhibit the relative Luciferase activity in cells. In addition, miR-101 overexpression markedly down-regulated the messenger RNA (mRNA) and protein expressions of Nrf2 in cells. MiR-101 plays an important role in regulating the proliferation and apoptosis of gastric mucosal epithelial cells by targeting the Nrf2-ARE signaling pathway.

Effect of green tea catechins on gastric mucosal dysplasia in insulin-gastrin mice.

Green tea catechins (GTCs) have been implicated in various physiological effects, including anti-carcinogenic activities. In the present study, we evaluated the effects of GTCs specifically on the development of gastritis and pre-malignant lesions in insulin-gastrin mice. Nine-week-old male INS-GAS mice (n=38) were supplemented with GTCs for 4 and 28 weeks, and their body weights, serum gastrin levels, histopathology and pro-inflammatory cytokine levels in gastric tissue and mucosal cell proliferation were monitored. Body weights of the GTC-treated mice were significantly lower than those of the untreated controls (P≤0.05). Serum gastrin levels were suppressed at the age of 37-weeks (P≤0.05). The histopathological scores indicated that the extent of dysplasia was significantly diminished (P≤0.05), although GTC supplementation did not affect the inflammation scores. The messenger RNA levels of interferon (IFN)-γ were significantly reduced at the age of 13 weeks (P≤0.05), although the changes did not reach statistical significance at the age of 37 weeks (P=0.056). The labeling index of Ki-67 immunohistochemistry was significantly decreased (P≤0.05). These results demonstrated that GTCs may play a protective role in the development of gastritis and pre-malignant lesions via an IFN-γ, gastrin, and mucosal cell proliferation-dependent mechanism in this rodent model and potentially in humans.

Su1973 Tamoxifen Induced Gastric Atrophy Is Regulated by the NF-κB Subunit NFKB1

Introduction Chronic Helicobacter felis infection of C57BL/6 mice results in gastric corpus atrophy. We have previously demonstrated that the outcome of H. felis infection is altered by deletion of specific NF-κB subunits. Nfkb1 -/- mice developed more severe gastric atrophy than wild-type (WT) mice 6 weeks after H. felis infection, whereas, Nfkb2 -/- mice were protected. The oestrogen receptor antagonist tamoxifen (TAM) has recently been shown to cause acute, reversible gastric atrophy in mice. In wild-type mice, this is associated with an increase in proliferating epithelial cells. We therefore hypothesised that gastric atrophy and altered epithelial cell turnover induced by TAM may be regulated by signalling involving NF-κB subunits. Method Groups of 5 adult female C57BL/6 (WT), Nfkb1 -/- , Nfkb2 -/- and c-Rel -/- mice were administered 150 mg/kg TAM by IP injection. Animals were culled 72 h later and the gastric corpus underwent quantitative histological assessment. Gastric atrophy was quantified using a validated visual analogue score. Cell positional scoring was used to quantify H + /K + ATPase expressing cells and Ki67 positive cells; gastric epithelial cells expressing cleaved-caspase 3 were quantified in 10 high-powered fields (hpf) per mouse. Results Morphological scoring demonstrated that TAM induced gastric atrophy in WT mice (median composite atrophy score (MCAS) 2 vs 0 in untreated WT mice, p + /K + ATPase or active caspase 3 positive cells was however seen. Nfkb2 -/- and c -Rel -/- mice exhibited similar phenotypes to WT mice. TAM treated Nfkb1 -/- mice developed more severe gastric atrophy with marked antralisation of the corpus epithelium (MCAS 4 vs 1 in untreated Nfkb1 -/- mice, p Nfkb1 -/- mice also exhibited corpus gland elongation after TAM treatment (36.1 ± 0.9 vs 23.9 ± 1.2cph, p 2.5 fold reduction in the number of parietal cells compared with untreated Nfkb1 -/- mice (p 2.3 fold increase in proliferating cells (p Nfkb1 -/- mice. Conclusion NFKB1 mediated signalling influences the severity of gastric mucosal atrophy and cell proliferation following TAM administration. This correlates closely with our previous observations following H. felis infection. As NFKB1 signalling plays a similar role in two models of gastric atrophy, this suggests that it may have a generic role in regulating the development of this pathology. Disclosure of interest None Declared.

Intestinal trefoil factor activates the PI3K/Akt signaling pathway to protect gastric mucosal epithelium from damage.

Intestinal trefoil factor (ITF, also named as trefoil factor 3, TFF3) is a member of the TFF-domain peptide family, which plays an essential role in the regulation of cell survival, cell migration and maintains mucosal epithelial integrity in the gastrointestinal tract. However, the underlying mechanisms and associated molecules remain unclear. The aim of this study was to explore the protective effects of ITF on gastric mucosal epithelium injury and its possible molecular mechanisms of action. In the present study, we show that ITF was able to promote the proliferation and migration of GES-1 cells via a mechanism that involves the PI3K/Akt signaling pathway. Western blot results indicated that ITF induced a dose- and time-dependent increase in the Akt signaling pathway. ITF also plays an essential role in the restitution of GES-1 cell damage induced by lipopolysaccharide (LPS). LPS induced the apoptosis of GES-1 cells, decreased cell viability significantly (P<0.01) and led to epithelial tight junction damage, which is attenuated via ITF treatment. The protective effect of ITF on the integrity of GES-1 was abrogated by inhibition of the PI3K/Akt pathway. Taken together, our results demonstrate that ITF promotes the proliferation and migration of gastric mucosal epithelial cells and preserves gastric mucosal epithelial integrity after damage is mediated by activation of the PI3K/Akt signaling pathway. This study suggested that the PI3K/Akt pathway could act as a key intracellular pathway in the gastric mucosal epithelium that may serve as a therapeutic target to preserve epithelial integrity during injury.

The expression level of TRAF1 in human gastric mucosa is related to virulence genotypes of Helicobacter pylori

Objective. To investigate the expression level of tumor necrosis factor receptor-associated factor 1 (TRAF1) in gastric mucosa tissue in patients infected with Helicobacter pylori (H. pylori) and to analyze the relationship between TRAF1 expression and H. pylori virulence. Methods. Gastric tissue samples were collected from patients with gastritis, atrophic gastritis, intestinal metaplasia with atypical hyperplasia, and gastric cancer. The expression level of TRAF1 in each group was analyzed by real-time polymerase chain reaction (PCR) and Western blot analysis. Virulence genotypes of H. pylori were determined by PCR. Results. Significant differences in TRAF1 mRNA levels were observed between the gastritis and gastric cancer groups, and the atrophic gastritis and gastric cancer groups (p < 0.05). Moreover, significant differences in TRAF1 protein levels were observed between the gastritis and intestinal metaplasia with atypical hyperplasia groups, between the gastritis and gastric cancer groups, and between the atrophic gastritis and gastric cancer groups (all p < 0.05). The virulence genotypes of cytotoxin-associated gene A (cagA), vacAs1, and vacAm1 were more frequent in the TRAF1 high-level group than in the TRAF1 low-level group (p < 0.05). Conclusion. Higher TARF1 expression level is associated with infection by CagA+/vacAs1+/m1+ virulent H. pylori strains and may promote the proliferation of gastric mucosal cells and induce gastric cancer.

A network map of the gastrin signaling pathway

Gastrin is the primary hormone that induces gastric acid secretion (Edkins 1906). In humans, the gene encoding for gastrin is located on chromosome 17q21 (Lund et al. 1986). This hormone is produced by the G-cells of the antrum of stomach as preprogastrin, which comprises of 101 amino acids and is cleaved between Ala-21 and Ser-22 to yield progastrin (Reeve et al. 1984). Progastrin is then sequentially cleaved by prohormone convertase and carboxypeptidase E to yield glycine-extended gastrins- a 35-amino acid gastrin-34-Gly (G34-Gly) or an 18-amino acid gastrin-17-Gly (G17-Gly) in endocrine cells (Varro et al. 1995; Lacourse et al. 1997). G34-Gly and G17-Gly are amidated at their carboxyl terminal groups by the enzyme peptidyl alpha-amidating mono-oxygenase to form amidated gastrin. Gastrin interacts with the membrane-bound G-protein coupled cholecystokinin receptor group consisting of Cholecystokinin A receptor (CCKAR) and Cholecystokinin B receptor (CCKBR). CCKAR possesses high affinity for cholecystokinin and has a negligible affinity for gastrin (Ferrand and Wang 2006; Dufresne et al. 2006). However, CCKBR has a high affinity for gastrin and their carboxyl amidated analogues. CCKBR is expressed in the brain, smooth muscle cells, and parietal cells (Kopin et al. 1992; Berna et al. 2007). In addition to this, gastrin has also been reported to interact with annexin 2, and thereby exerts proliferation effects in gastrointestinal cancers (Singh et al. 2007; Singh 2007; Sarkar et al. 2012). The release of gastrin is induced by gastrin-releasing peptide, a neurotransmitter which acts on its basolateral receptor in the G-cells. Binding of gastrin to CCKBR present in parietal and enterochromaffin-like (ECL) cells (Schmitz et al. 2001; Kulaksiz et al. 2000)induces gastric acid secretion by parietal cells as well as histamine release by ECL cells (Dockray et al. 2005). The released histamine reaches parietal cells by paracrine diffusion where it binds H2 receptors and induces gastric acid secretion. The secretion of gastric acid inhibits the release of gastrin hormone rendering a negative feedback control, further preventing excess acid secretion. In addition, low pH value in the stomach inhibits gastrin release through stimulating somatostatin secretion by antral D-cells (Bloom et al. 1974). Gastrin is a well-known growth factor for the gastrointestinal tract. Gastrin stimulates proliferation of gastric mucosal cells (Hansen et al. 1976), maturation of parietal cells and enterochromaffin-like cells (Jain and Samuelson 2006), and promotes islet differentiation in the pancreas (Wang et al. 1993). It has also been shown to stimulate proliferation of gastric (Ishizuka et al. 1992) and colon cancer cells (Watson et al. 1989). Gastrin modulates invasion (Wroblewski et al. 2002), apoptosis (Przemeck et al. 2008; Todisco et al. 2001) and migration (Noble et al. 2003) in epithelial cells. Both glycine-extended gastrin and amidated gastrin have been reported to induce angiogenesis (Clarke et al. 2006; Lefranc et al. 2004). Moreover, the hypergastrinemic state has been associated with diverse physiological disorders in humans and other mammals. These include atrophic gastritis (Lehy et al. 2000), pernicious anemia (Orlando et al. 2007), excess acid secretion leading to duodenal ulcer disease in Helicobacter pylori infection (Berna et al. 2006; Jensen 2002; Scarpignato et al. 1996) and gastrinoma (Kloppel and Anlauf 2007). Recently, a number of research groups have explored the feasibility of using gastrin to treat various diseases. Gastrin has been used in combination therapy with epidermal growth factor to increase beta-cell mass which reversed hyperglycemia in diabetic mice (Suarez-Pinzon et al. 2005). Gastrin-stimulated beta cell neogenesis when used in combination with glucagon-like peptide 1 in human pancreatic duct cell transplanted in immunodeficient diabetic mice (Suarez-Pinzon et al. 2008). Gastrins and their receptors have been suggested as potential targets to treat gastrointestinal and pancreatic cancers (Rengifo-Cam and Singh 2004). Gastrin has also been shown to have therapeutic promise as it inhibits the growth of cholangiocarcinoma cells by promoting apoptosis (Kanno et al. 2001). On account of the functional significance of different natural forms of gastrin, we have assembled signaling pathway reactions induced by them upon binding to gastrin receptor(s) in different human cell types. These manually curated signaling events are made available through NetPath (http://www.netpath.org/) (Kandasamy et al. 2010) in different formats for analysis by the scientific community.

Index of Suspicion

The reader is encouraged to write possible diagnoses for each case before turning to the discussion. We invite readers to contribute case presentations and discussions. Please inquire first by contacting Dr. Deepak Kamat at DKamat@med.wayne.edu. A 2-year-old boy with a history of eczema and food allergies to nuts and soy presents to the emergency department with a 10-day history of generalized edema. The swelling began in the periorbital area but soon progressed to involve his entire body. There have been no fevers or any recent illnesses other than 1 day of cough and a few watery stools. He has had no new exposures to foods or other substances. His family history is remarkable for multiple allergies and eczema. Physical examination reveals a temperature of 98.8°F (37.1oC), heart rate of 98 beats per minute, respiratory rate of 36 breaths per minute, blood pressure of 103/65 mm Hg, and an oxygen saturation of 97% on room air. His weight is 12.6 kg (34th percentile). He is noted to have a moderate amount of periorbital edema, decreased breath sounds at the lung bases bilaterally, a protuberant abdomen, and 2+ pitting edema on his lower extremities. The rest of his physical examination findings are within normal limits. Laboratory evaluation reveals a white blood cell count of 23,800/μL (23.8 × 109/L), with 37% neutrophils, 49% lymphocytes, 4% bands, 7% monocytes, and 3% eosinophils; hemoglobin level of 13.3 g/dL (133 g/L); and platelet count of 427 × 103/μL (427 × 109/L). Serum electrolyte, blood urea nitrogen, and creatinine levels are normal. Sodium level is 132 mEq/L (132 mmol/L). Serum albumin level is 1.7 g/dL (17 g/L) (reference range, 3.4-5.4 g/dL [34-54 g/L]). Total protein level is 3.8 g/dL (38 g/L). Aspartate aminotransferase, alanine aminotransferase, and coagulation profiles …

α-Tocopherol administration blocks adaptive changes in cell NADH/NAD+ redox state and mitochondrial function leading to inhibition of gastric mucosa cell proliferation in rats

In experimentally induced chronic gastritis, a compensatory mucosal cell proliferation occurs with enhanced glucose oxidative metabolism linked to lipoperoxidative events. Therefore, this study was aimed at assessing the participation of cell NAD/NADH redox state and mitochondrial functions during gastric mucosa proliferation and the effects of in vivo α-tocopherol (vitamin E) administration. Glucose oxidation and oxygen consumption were tested in gastric mucosa samples obtained from rats with gastritis and from those also treated with α-tocopherol. Gastric mucosal mitochondria were isolated and structural and functional parameters were determined. Succinate oxidation, ADP phosphorylation, mitochondrial enzyme activities, and membrane lipid composition were measured. In addition, parameters indicative of cellular NAD/NADH redox state, proliferation, apoptosis, and nitric oxide (NO) metabolism were also determined. After ethanol withdrawal, the damaged gastric mucosa increased glucose and oxygen consumption, events associated with a more reduced cytoplasmic NAD/NADH ratio. Enhanced mitochondrial oxidative phosphorylation and increased mitochondrial enzyme activities occurred early, accompanied by recovery of lost mitochondrial protein and lipid composition in the gastric mucosa, events associated with increased NO production. When mitochondrial function and structural events were normalized, apoptosis was initiated as assessed by the mitochondrial Bax/Bcl2 ratio. Treatment with α-tocopherol inhibited cell proliferation and blocked enhanced glucose utilization, mitochondrial substrate oxidation, and changes in redox state, delaying the onset of these adaptive metabolic changes, whereas it inhibited cell proliferation. In conclusion, α-tocopherol could abolish damage-induced “stress” signaling by desynchronizing mitochondrial adaptive responses, including mitochondria biogenesis, and consequently NAD/NADH redox, which seems to regulate gastric mucosal cell proliferation.

Lateral hypothalamic area mediated the protective effects of microinjection of glutamate into interpositus nucleus on gastric ischemia-reperfusion injury in rats

We investigated the protective effects of chemical stimulation of cerebellar interpositus nucleus (IN) on gastric ischemia-reperfusion injury (GI-RI) and its possible regulatory mechanisms in rats. Gastric mucosal damage index (GMDI) indicated the severity of gastric mucosal injuries. Transferase dUTP nick end labeling (TUNEL) staining and proliferating cell nuclear antigen (PCNA) were performed to assess gastric mucosal cell apoptosis and proliferation. Microinjection of glutamate into IN markedly attenuated GI-RI. Either chemical lesion of IN or electrical ablation of the decussation of superior cerebellar peduncle (DSCP) obviously aggravated GI-RI. The protective effects of IN were reversed with the pretreatments of microinjection of 3-mercaptopropionic acid into IN or Bicuculline into lateral hypothalamic area (LHA), individually. The discharge frequency and intensity of greater splanchnic nerve (GSN) decreased and gastric mucosal blood flow increased after chemical stimulation of IN. The apoptosis of positive cells of gastric mucosa was decreased by chemical stimulation of IN, whereas proliferation increased. The gastric juice volume, acidity, and total acid output were all decreased after the chemical stimulation of IN. These results indicated that IN participates in regulation of GI-RI and is a specific area in central nervous system for exerting protective effects on GI-RI. DSCP, LHA and GSN may involve in this process. Apoptosis and proliferation may mediate this protective process in rats too.

ASSESSMENT OF WOUND HEALING ACTIVITY OF DAPHNE OLEOIDES SUBSP. KURDICA: IDENTIFICATION OF THE ACTIVE COMPONENTS USING BIOASSAY GUIDED ISOLATION METHOD

s. PHYTOPHARM 2012 M 76 Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] ТОМ 10/2012/2 CRS) and aspirin (4h ASP) induced ulceration. The ulcer index and % protection was calculated. The gastric juice collected from 4h Pl rats was studied for the effects on gastric volume, ph, free acid, total acid-pepsin output, dNA gastric juice cell shedding and mucosal cell proliferation. biochemical estimation such as histamine content, mucus content and hkATPase inhibitory activity was also evaluated. The effect of APf on stomach histopathology was also studied. Statistical analysis was done by one way ANOVA followed by dunnett’s post hoc test for multiple comparisons against control group. The difference was considered to be significant when P < 0.05. Oral treatment of APf (200 and 400 mg/kg, p. o.) for 7 days showed significant protection against acute gastric ulcer induced by 4h Pl and 95 % EtOh, but was found to have no effect on CRS and ASP-induced gastric ulceration. The gastric studies showed that treatment with APf signiccantly increased the gastric juice ph, decrease the gastric volume and total acid-pepsin output and significantly reduced the gastric juice cell shedding. A significant decrease in histamine content and a reduction in free phosphate ions responsible for hkATPase inhibitory activity were also observed. in conclusion, the results of the present study indicate that the gastroprotective activity of APf contributes mainly to the peripheral effects on the gastric secretion and the probable mechanism underlying behind such activity might be both due to the possible inhibition on the histaminic (h 2 ) receptor and hkATPase pump. INVESTIGATION OF THREE TRITERPENE ACIDS AGAINST CANCER CELL PROLIFERATION ACTIVITY © Khoo Gaik Ming, Clausen Morten Rahr, Larsen Erik Department of Food Science, Aarhus University, Kirstinebjergvej 10, DK-5792 Aarslev, Denmark The object of this study was to investigate the inhibitory effect of betulinic acid, oleanolic acid and ursolic acid on human colon cancer cell proliferation activity. Triterpene acids are members of the terpene family and are widely distributed in food plants. A rosehip fraction containing a mixture of betulinic acid, oleanolic acid and ursolic acid displayed inhibitory effect against the release of pro-inflammatory cytokine interleukin-6 with the combination of oleanolic acid/betulinic acid and ursolic acid/betulinic acid displaying significant synergistic inhibitory effects (1). Our results showed that these three compounds also displayed good inhibitory effect against cancer cell proliferation activity at the concentration of 10μg/ml. both betulinic acid and ursolic acid displayed strong inhibition against hT-29 (88 % and 79 %, respectively) and SW-480 (76 % and 82 %, respectively), but a moderate to weak inhibition against Caco-2 (19 % and 22 %, respectively). Oleanolic acid displayed only moderate inhibition against Caco-2 (49 %), very weak inhibition against hT-29 (7 %) and no inhibition against SW-480. No synergistic effects were observed between combinations of the triterpene acids. We only found an additive effect in the combination of oleanolic acid/ursolic acid and only against proliferation of hT-29. Antagonistic effects between the pure compounds were observed when betulinic acid was one of the components in the combination. Although betulinic acid and ursolic acid showed good inhibitory effects against all three cancer cell lines, their combination displayed antagonistic effects and showed poor inhibition against cancer cell proliferations. however, these triterpenic acids and their combination effects should be further investigated in different in vitro or in vivo experiment to confirm their bioactivities and mechanism. Reference: (1) Saaby l., Moesby l., hansen E. W., Christensen S. b. (2011). isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina l.). Phytotherapy Research, 25, 195–201. DECURSIN AND RELATED COMPOUNDS INHIBIT VASCULAR ENDOTHELIAL MIGRATION BY SPHINGOSINE KINASE INHIBITION © Lee Yong-Moon, Shin Kyong-Oh, Seo Cho-Hee, Kim Tack-Joong College of Pharmacy, Chungbuk National University, Cheongju, South Korea Division of Biological Science and Technology, Yonsei University, Wonju, South Korea The pyranocoumarin compound decursin isolated from the herb, Angelica gigas, are known to possess potent anti-inflammatory activities. however, little is known about their anti-angiogenic activity. here, we investigated the anti-angiogenic effects of decursin by the changes of sphingolipid metabolites

INVESTIGATION OF THREE TRITERPENE ACIDS AGAINST CANCER CELL PROLIFERATION ACTIVITY

s. PHYTOPHARM 2012 M 76 Obzory po kliniceskoj farmacologii i lekarstvennoj terapii [Reviews of clinical pharmacology and drug therapy] ТОМ 10/2012/2 CRS) and aspirin (4h ASP) induced ulceration. The ulcer index and % protection was calculated. The gastric juice collected from 4h Pl rats was studied for the effects on gastric volume, ph, free acid, total acid-pepsin output, dNA gastric juice cell shedding and mucosal cell proliferation. biochemical estimation such as histamine content, mucus content and hkATPase inhibitory activity was also evaluated. The effect of APf on stomach histopathology was also studied. Statistical analysis was done by one way ANOVA followed by dunnett’s post hoc test for multiple comparisons against control group. The difference was considered to be significant when P < 0.05. Oral treatment of APf (200 and 400 mg/kg, p. o.) for 7 days showed significant protection against acute gastric ulcer induced by 4h Pl and 95 % EtOh, but was found to have no effect on CRS and ASP-induced gastric ulceration. The gastric studies showed that treatment with APf signiccantly increased the gastric juice ph, decrease the gastric volume and total acid-pepsin output and significantly reduced the gastric juice cell shedding. A significant decrease in histamine content and a reduction in free phosphate ions responsible for hkATPase inhibitory activity were also observed. in conclusion, the results of the present study indicate that the gastroprotective activity of APf contributes mainly to the peripheral effects on the gastric secretion and the probable mechanism underlying behind such activity might be both due to the possible inhibition on the histaminic (h 2 ) receptor and hkATPase pump. INVESTIGATION OF THREE TRITERPENE ACIDS AGAINST CANCER CELL PROLIFERATION ACTIVITY © Khoo Gaik Ming, Clausen Morten Rahr, Larsen Erik Department of Food Science, Aarhus University, Kirstinebjergvej 10, DK-5792 Aarslev, Denmark The object of this study was to investigate the inhibitory effect of betulinic acid, oleanolic acid and ursolic acid on human colon cancer cell proliferation activity. Triterpene acids are members of the terpene family and are widely distributed in food plants. A rosehip fraction containing a mixture of betulinic acid, oleanolic acid and ursolic acid displayed inhibitory effect against the release of pro-inflammatory cytokine interleukin-6 with the combination of oleanolic acid/betulinic acid and ursolic acid/betulinic acid displaying significant synergistic inhibitory effects (1). Our results showed that these three compounds also displayed good inhibitory effect against cancer cell proliferation activity at the concentration of 10μg/ml. both betulinic acid and ursolic acid displayed strong inhibition against hT-29 (88 % and 79 %, respectively) and SW-480 (76 % and 82 %, respectively), but a moderate to weak inhibition against Caco-2 (19 % and 22 %, respectively). Oleanolic acid displayed only moderate inhibition against Caco-2 (49 %), very weak inhibition against hT-29 (7 %) and no inhibition against SW-480. No synergistic effects were observed between combinations of the triterpene acids. We only found an additive effect in the combination of oleanolic acid/ursolic acid and only against proliferation of hT-29. Antagonistic effects between the pure compounds were observed when betulinic acid was one of the components in the combination. Although betulinic acid and ursolic acid showed good inhibitory effects against all three cancer cell lines, their combination displayed antagonistic effects and showed poor inhibition against cancer cell proliferations. however, these triterpenic acids and their combination effects should be further investigated in different in vitro or in vivo experiment to confirm their bioactivities and mechanism. Reference: (1) Saaby l., Moesby l., hansen E. W., Christensen S. b. (2011). isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina l.). Phytotherapy Research, 25, 195–201. DECURSIN AND RELATED COMPOUNDS INHIBIT VASCULAR ENDOTHELIAL MIGRATION BY SPHINGOSINE KINASE INHIBITION © Lee Yong-Moon, Shin Kyong-Oh, Seo Cho-Hee, Kim Tack-Joong College of Pharmacy, Chungbuk National University, Cheongju, South Korea Division of Biological Science and Technology, Yonsei University, Wonju, South Korea The pyranocoumarin compound decursin isolated from the herb, Angelica gigas, are known to possess potent anti-inflammatory activities. however, little is known about their anti-angiogenic activity. here, we investigated the anti-angiogenic effects of decursin by the changes of sphingolipid metabolites

Transmural Pressure Loading Enhances Gastric Mucosal Cell Proliferation

Although increased intraluminal pressure in the stomach due to gastric outlet obstruction or functional gastric motor dysfunction, including gastroparesis, may affect gastric mucosal integrity, the direct effect of mechanical pressure on gastric mucosal cells has not yet been fully investigated. The aims of this study were to determine whether exposure to transmural pressure would affect the proliferation of gastric mucosal cells and to elucidate the intracellular signaling pathways involved. Cellular proliferation and DNA synthesis were evaluated in rat gastric epithelial cells exposed to high transmural pressures. The levels of activation of 3 MAP kinases, ERK, JNK, and p38, were assessed, and the induction of immediate early gene expression was examined. The activation of nuclear factor activator protein-1 (AP-1) was evaluated by an electrophoretic mobility shift assay. Exposure to high transmural pressure significantly increased DNA synthesis within 24 h, with the most marked increase observed after exposure to a pressure of 80 mmHg, and this increase was inhibited by the MEK1 inhibitor PD98059. Early activation of ERK kinase, but not of JNK or p38 kinase, was detected after pressure loading. Early induction of the c-fos and c-myc genes and activation of the AP-1 transcription factor were also demonstrated within 3 h of exposure to 80 mmHg of pressure. Gastric mucosal cell proliferation induced by exposure to high transmural pressure may be related to early activation of ERK, the induction of c-fos and c-myc, and the activation of AP-1.

Cell proliferation in ventromedial hypothalamic lesioned rats inhibits acute gastric mucosal lesions

AimThe role of mucosal layer thickness on prevention of acute gastric mucosal lesions (AGMLs) was examined in ventromedial hypothalamic (VMH)-lesioned rats. Materials and methodsThe incidence of AGMLs after 48-h fasting and 60% ethanol injection into the stomach after 24-h fasting, aggressive factors (gastric acid and serum gastrin) and defensive factors [hexosamine, gastric mucosal blood flow (GMBF), serum thiobarbituric acid reacting substances (TBARS), and thickness of the gastric mucosal layer] were evaluated in VMH-lesioned rats. The effects of cell proliferation on the gastric mucosal layer of these rats were evaluated by H–E staining and immunostaining with proliferating cell nuclear antigen (PCNA). ResultsAfter 48-h fasting, no AGMLs were observed in VMH-lesioned and sham VMH-lesioned rats (controls). With 60% ethanol administration after 24-h fasting, the numbers of AGMLs were similar in the two groups, but the ulcer index, a marker of ulcer formation, was lower in VMH-lesioned rats compared to that in sham VMH-lesioned rats. VMH-lesioned rats showed increased gastric acid secretion and serum gastrin compared to sham VMH-lesioned rats, indicating an increase in aggressive factors in VMH-lesioned rats. The two groups had similar levels of gastric mucosal hexosamine, GMBF, and gastric mucosal TBARS, but VMH-lesioned rats had an increased thickness of the mucosal cell layer, indicating an increase in defensive factors in these rats. Histologically, VMH-lesioned rats had an increased total mucosal cell layer, especially for the surface epithelial cell layer, and an increased PCNA-labeling index, a marker of cell proliferation, especially in the proliferative zones of gastric mucosa, indicating increased cell proliferation in the proliferative zone of the gastric mucosa. ConclusionVMH-lesioned rats are resistant to AGML formation due to increased cell proliferation in gastric mucosa through elevating the levels of defensive factors over those of aggressive factors.

Effect of Bone Morphogenetic Protein-2 on Proliferation and Apoptosis of Gastric Cancer Cells

Objective: To investigate the effects of bone morphogenetic protein-2 (BMP-2) on the proliferation, differentiation and apoptosis of normal human gastric mucosal cells and gastric cancer cells.Methods: Poorly differentiated gastric cancer BGC823 cells, moderately differentiated gastric cancer cells and normal human gastric mucosal epithelial GES-1 cells were independently treated with recombinant human BMP-2 or its inhibitor Noggin. MTT assay was performed to detect the proliferation, flow cytometry done to measure the cell cycle and apoptosis and immunohistochemistry carried out to determine the expression of cyclin-dependent kinase 4 (CDK4).Results: BMP-2 exerted inhibitory effect on the growth of all types of cells and the inhibition become more evident with the increase of BMP-2 dose. After treatment with 200 ng/ml BMP-2, cancer cells arrested in G1 phase and those in S phase reduced. Gastric cancer cells had higher CDK4 expression than GES-1 cells. BMP-2 decreased CDK-4 expression in cancer cells but had no influence in GES-1 cells. Noggin conferred promotive effect on the growth of 3 types of cells. In 2 types of cancer cells, treatment with 2000 ng/ml Noggin significantly increased the proportion of cells in S phase but reduced that in G1 phase. However, Noggin did not affect the cell cycle of GES-1 cells. The CDK4 expression was markedly increased in 2 types of cancer cells but that of GES-1 remained unchanged after treatment with 2000 ng/ml Noggin.Conclusions: BMP-2 may inhibit the proliferation of both normal and malignant gastric epithelial cells, down-regulate CDK4 expression in gastric cancer cells and arrest gastric cancer cells in G1-phase in cell cycle. Through antagonizing BMP-2, Noggin, may accelerate the proliferation of gastric cancer cells. Thus, the abnormality of BMP signaling pathway may play an important role in the pathogenesis of gastric cancer.