Mucosal Homogenates Research Articles

Protective effect of dimethyl fumarate against ethanol-provoked gastric ulcers in rats via regulation of HMGB1/TLR4/NF-κB, and PPARγ/SIRT1/Nrf2 pathways: Involvement of miR-34a-5p

Aberration of the gastric mucosal barrier homeostasis circuit is one of the key features linked to the onset of gastric ulcers (GU). This work aimed to inspect the gastroprotective influence of dimethyl fumarate (DMF) on ethanol-induced GU in rats and to decipher the possible mechanisms entailed. Rats were pretreated with either DMF (80 mg/kg) or omeprazole (OMP) (20 mg/kg) by oral gavage for 2 weeks. After 24 h of starvation, ethanol (5 ml/kg, oral) was employed to trigger GU in rats, while carboxymethyl cellulose (CMC) was used as a control. Ethanol notably elevated both macroscopic and microscopic gastric damage. DMF and OMP exhibited similar effects on gastric ulcer healing. DMF intervention led to a substantial improvement in gastric insults. DMF significantly reduced ethanol-triggered gastric lesions, as manifested by decreased gastric secretion, acidity, ulcer surface area percent, reduced leukocyte incursion, and increased mucus percent. DMF upregulated miR-34a-5p expression concomitant with the suppression of high mobility group box1 (HMGB1) and inflammatory responses in gastric mucosal homogenate. DMF improved GU by restoring reduced antioxidant defense mechanisms through the coactivation of nuclear factor erythroid 2-related factor-2 (Nrf2), peroxisome proliferator-activated receptor gamma (PPARγ), and sirtuin1 (SIRT1), indicating the protective role of the PPARγ/SIRT1/Nrf2 pathway. Intriguingly, DMF mitigated apoptosis in ethanol-elicited GU. Taken together, this research implies the potential for the repurposing of DMF as an innovative gastroprotective medication to reestablish the balance of the gastric mucosal barrier via the attenuation of gastric inflammation, oxidative stress, and apoptosis.

Genetic and acquired sucrase-isomaltase deficiency: A clinical review.

Genetic sucrase-isomaltase deficiency (GSID) is an inherited deficiency in the ability to digest sucrose and potentially starch due to mutations in the sucrase-isomaltase (SI) gene. Congenital sucrase-isomaltase deficiency is historically considered to be a rare condition affecting infants with chronic diarrhea as exposure to dietary sucrose begins. Growing evidence suggests that individuals with SI variants may present later in life, with symptoms overlapping with those of irritable bowel syndrome. The presence of SI genetic variants may, either alone or in combination, affect enzyme activity and lead to symptoms of different severity. As such, a more appropriate term for this inherited condition is GSID, with a recognition of a spectrum of severity and onset of presentation. Currently, disaccharidase assay on duodenal mucosal tissue homogenates is the gold standard in diagnosing SI deficiency. A deficiency in the SI enzyme can be present at birth (genetic) or acquired later, often in association with damage to the enteric brush-border membrane. Other noninvasive diagnostic alternatives such as sucrose breath tests may be useful but require further validation. Management of GSID is based on sucrose and potentially starch restriction tailored to the individual patients' tolerance and symptoms. As this approach may be challenging, additional treatment with commercially available sacrosidase is available. However, some patients may require continued starch restriction. Further research is needed to clarify the true prevalence of SI deficiency, the pathobiology of single SI heterozygous mutations, and to define optimal diagnostic and treatment algorithms in the pediatric population.

Anti H. pylori, anti-secretory and gastroprotective effects of Thymus vulgaris on ethanol-induced gastric ulcer in Sprague Dawley rats.

The objectives of the present study were to evaluate the acute toxicity, gastroprotective, therapeutic, anti-inflammatory and anti H. pylori activities of T. vulgaris total plant extract against ethanol-induced gastric ulcers in Sprague Dawley rats. Animals were divided into five groups i.e G-1 (Normal Control), Group 2 (ulcer control) were administered orally with 0.5% Carboxymethylcellulose (CMC), Group 3 (omeprazole treated) was administered orally with 20 mg/kg of omeprazole and Groups 4 and 5 (Low dose and High dose of the extract) were administered orally with 250, and 500 mg/ kg of Thymus vulgaris extract, respectively. After 1 hour, the normal group was orally administered with 0.5% CMC (5 ml/kg), whereas absolute alcohol (5ml/ kg) was orally administered to the ulcer control group, omeprazole group, and experimental groups. Stomachs were examined macroscopically and microscopically. Grossly, rats pre-treated with T. vulgaris demonstrated significantly decreased ulcer area and an increase in mucus secretion and pH of gastric content compared with the ulcer control group. Microscopy of gastric mucosa in the ulcer control group showed severe damage to gastric mucosa with edema and leukocytes infiltration of the submucosal layer. However, rats pretreated with omeprazole or Thyme vulgaris exhibited a mild to moderate disruption of the surface epithelium and lower level of edema and leukocyte infiltration of the submucosal layer. The T. vulgaris extract caused up-regulation of Hsp70 protein, down-regulation of Bax protein, and intense periodic acid Schiff uptake of the glandular portion of the stomach. Gastric mucosal homogenate of rats pre-treated with T. vulgaris exhibited significantly increased superoxide dismutase (SOD) and catalase (CAT) activities while malondialdehyde (MDA) level was significantly decreased. Based on the results showed in this study, Thymus vulgaris extract can be proposed as the safe medicinal plants for use and it has considerable gastroprotective potential via stomach epithelium protection against gastric ulcers and stomach lesions.

Study of the antiulcer activity of garden cabbage extract on the chronic acetic ulcer model in rats

The aim of the study was to study the gastroprotective properties of the dry extract of garden cabbage in the model of chronic acetic ulcer in rats.
 Materials and methods. Chronic ulcerative lesions of the gastric mucosa of rats were modeled by introducing 0.05 ml of a 30 % solution of acetic acid into the subserous layer of the stomach wall. The dry extract of garden cabbage in a conditionally therapeutic dose of 50 mg/kg and reference drugs omeprazole and altan were administered intragastrically for 10 days, starting from the second day of the experiment. When choosing the duration of the course of treatment, we proceeded from the data that the effectiveness of the antiulcer action in the clinic is assessed after 10-12 days of treatment. The percentage of animals with ulcers in the group, the state of the gastric mucosa were evaluated, the ulcer index and antiulcer activity were calculated. To elucidate the possible mechanism of action of the extract, we studied its effect on synthetic processes in terms of the content of RNA and DNA in homogenates of the gastric mucosa by the spectrophotometric method by reaction with perchloric acid.
 Results. On the model of stomach damage, caused by acetic acid, a pronounced antiulcer activity of the extract (54.26) was established at the level of omeprazole (54.62). The maximum antiulcer activity (72.75) was revealed when using the combination of the extract with omeprazole, which, in terms of ulcerative effect, is superior to monotherapy with the extract, omeprazole and altan (35.08).
 Reparative regeneration is a component of an adaptive tissue reaction; under conditions of violation of the reparative potencies of the mucosa, chronic diseases of the gastrointestinal tract develop. Since the final goal of antiulcer therapy is the healing of the ulcer, and the basis of this process is reparative regeneration, the reparative properties of the extract have been studied. Under the influence of cabbage extract, the content of RNA and DNA in the mucosa probably increased by 2.8 and 2.4 times, respectively, at the same level with omeprazole. If possible, to activate the synthetic processes of DEGC and omeprazole, altan significantly prevailed.
 The most pronounced effect of enhancing reparative processes was established with the combined use of the extract and omeprazole, which turned out to be a significant increase in the content of RNA and DNA in the gastric mucosa by 3.2 and 2.9 times compared with the control.
 Conclusions. On the model of chronic damage to the stomach of rats, cabbage extract showed an antiulcer effect, the severity of which was not inferior to the comparison drug, the proton pump inhibitor omeprazole, and significantly exceeded the phytopreparation - altan. The combined use of the extract and omeprazole showed the highest preventive effect of preventing the negative effects of acetic acid on the gastric mucosa. One of the mechanisms of the gastroprotective action of the extract is its ability to enhance reparative processes in the gastric mucosa. The results obtained indicate the prospects for further research on the antiulcer properties of cabbage extract with a view to its use in the complex therapy of peptic ulcer

Oral administration of Nigella sativa oil attenuates arsenic-induced redox imbalance, DNA damage, metabolic distress, and histopathological alterations in rat intestine

BackgroundExposure to arsenic, a widespread environmental toxin, produces multiple organ toxicity, including gastrointestinal toxicity. Nigella sativa (NS) has long been revered for its numerous health benefits under normal and pathological states. In view of this, the present study attempts to evaluate the protective efficacy of orally administered Nigella sativa oil (NSO) against arsenic-induced cytotoxic and genotoxic alterations in rat intestine and elucidate the underlying mechanism of its action. MethodsRats were categorized into the control, NaAs, NSO, and NaAs+NSO groups. After pre-treatment of rats in the NaAs+NSO and NSO groups daily with NSO (2 ml/kg bwt, orally) for 14 days, NSO treatment was further continued for 30 days, with and without NaAs treatment (5 mg/kg bwt, orally), respectively. Various biochemical parameters, such as enzymatic and non-enzymatic antioxidants, carbohydrate metabolic and brush border membrane marker enzyme activities were evaluated in the mucosal homogenates of all the groups. Intestinal brush border membrane vesicles (BBMV) were isolated, and the activities of membrane marker enzyme viz. ALP, GGTase, LAP, and sucrase were determined. Further, the effect on kinetic parameters viz KM (Michaelis-Menten constant) and Vmax of these enzymes was assessed. Integrity of enterocyte DNA was examined using the comet assay. Histopathology of the intestines was performed to evaluate the histoarchitectural alterations induced by chronic arsenic exposure and/or NSO supplementation. Arsenic accumulation in the intestine was studied by inductively coupled plasma-mass spectroscopy (ICP-MS). ResultsNaAs treatment caused substantial changes in the activities of brush border membrane (BBM), carbohydrate metabolism, and antioxidant defense enzymes in the intestinal mucosal homogenates. The isolated BBM vesicles (BBMV) also showed marked suppression in the marker enzyme activities. Severe DNA damage and mucosal arsenic accumulation were observed in rats treated with NaAs alone. In contrast, oral NSO supplementation significantly alleviated all the adverse alterations induced by NaAs treatment. Histopathological examination supported the biochemical findings. ConclusionNSO, by improving the antioxidant status and energy metabolism, could significantly alter the ability of the intestine to protect against free radical-mediated arsenic toxicity in intestine. Thus, NSO may have an excellent scope in managing gastrointestinal distress in arsenic intoxication.

Factors Affecting Endogenous Oxalate Synthesis and Its Excretion in Feces and Urine in Rats

It has been observed that the feces as well as urine of rats fed diets supplemented with 3% glycine and 5.2% hydroxyproline contain unexpectedly high amounts of endogenously formed oxalate. That intestinal microorganisms do not synthesize significant amounts of oxalate was indicated by the findings that oral tetracycline had no effect on oxalate excretion and that germ-free rats excreted more oxalate than conventional rats. Since little intraperitoneally injected [14C]oxalate appeared in the feces, and rat intestinal mucosa homogenates were found to produce oxalate from a variety of precursors of which glyoxylic acid was by far the most important, it is probable that the intestinal mucosa may be an important source of fecal oxalate observed in these studies. Ninety percent of weanling rats fed complete diets supplemented with glycine and hydroxyproline developed urinary stones in 38 days. It has been concluded that in the treatment of patients with histories of calcium oxalate urolithiasis, more concern than is commonly shown should be directed towards the feeding of diets high in precursors of endogenous oxalate synthesis.oxalate hydroxyproline calculi

Feeding Colostrum Rapidly Alters Enzymatic Activity and the Relative Isoform Abundance of Jejunal Lactase in Neonatal Pigs ,

Our objective was to determine the short-term effects of feeding colostrum and mature milk on the enzyme activity and relative isoform abundance of lactase in neonatal pigs. We studied newborn pigs that were fed colostrum, mature milk and water for 6 h (Study 1). We also studied unfed pigs shortly after birth, and newborn pigs fed either colostrum or mature milk for 24 h (Study 2). The specific activity (micromol x min(-1) x g protein(-1)) of lactase in ajejunal mucosal homogenate and a purified membrane fraction was lower in pigs fed colostrum than in unfed newborns or those fed either milk or water. However, after 24 h, total jejunal lactase activity (micromol x mol(-1)), jejunal mass and protein content were higher in the colostrum- and milk-fed pigs than in the unfed newborns. In colostrum-fed pigs, the reduction in lactase specific activity after 6 h was associated with 1) a marked increase in the relative abundance of a 180-kDa protein, which was shown to be one of three pro-lactase isoforms, and 2) a lower relative abundance of the 160-kDa isoform, considered to be the mature form of the enzyme. Our evidence suggests that feeding either colostrum or mature milk increases total jejunal lactase activity. The reduction in both the specific activity and abundance of the mature isoform in conjunction with an increased relative abundance of the 180-kDa pro-lactase isoform suggests that feeding colostrum alters the post-translational processing of intestinal lactase in neonatal pigs.

CHARACTERISTICS OF CYTOPROTECTIVE EFFECT ON THE GASTRIC MUCOSA OF CRYOPRESERVED PLACENTA EXTRACT UNDER WATER-IMMOBILIZATION STRESS

Various stressors are currently considered to play a significant role among the etiological factors for development of peptic ulcer disease. Along with psycho-emotional stressing experiences, the causes of formation of stress ulcers include major surgical interventions, thermal or mechanical injuries, severe blood loss, progressive syndrome of endogenous intoxication and more. Among patients in intensive care units, the incidence of stress ulcers is 5.0%, and in 30.0-50.0% of cases stress ulcers are complicated by bleeding and perforation.
 Aim. The research aims at characterizing the gastrocytoprotective effect of cryopreserved placenta extract (CEP) on the model of water-immobilization stress in rats according to macroscopic data and the results of biochemical study of homogenates of the gastric mucosa (GM).
 Materials and Methods. In vivo experimental studies were performed on 28 nonlinear laboratory male rats weighing 200-220 g. Stress-induced gastric ulcer was simulated under water-immobilization stress (WIS) in rats. WIS simulation according to the method of Takagi K.Y. et al. The activity of NO synthases (NOS) in GM was determined spectrophotometrically by the amount of oxidized NADPH+H+.
 Results and Discussion. The use of CEP, as well as esomeprazole, led to a weakening of GM ulcers. This was indicated by a statistically significant (p<0.05) decrease in ulcer index (UI) relative to the indicators of control rats by 9.8 and 3.3 times, respectively. At the same time, this indicator was three times lower on the background of preventive use of CEP than in rats treated with esomeprazole, respectively, antiulcer activity (AUA) was 96.4% and 69.2%. The study of the activity of constitutive (cNOS) and inducible (iNOS) isoforms of NOS showed that at the pathobiochemical level against the background of stress-induced ulcerogenesis in the tissues of the central nervous system there is a statistically significant (p<0.001) increase in iNOS activity in 5.2 times was 0.89±0.01 NADPH2/min´g protein. According to the degree of modulation of the activity of both total NOS and its individual isoforms, the study showed that preventive five-day administration of esomeprazole is inferior to the effectiveness of CEP. Thus, the activity of total NOS in rats administered esomeprazole was statistically significantly (p<0.001) decreased by only 17.1%, while the activity of this enzyme decreased (p<0.001) by 35.8% due to the use of CEP.
 Conclusions. Prophylactic five-day administration of CEP modulated the activity of the NOS system in the central nervous system, which in a macroscopic study appeared statistically significant (p<0.05) reduction in UI relative to rats in the control group by 9.8 times. Thus, NOS activity statistically significantly (p<0.001) decreased by 35.8%, and iNOS activity statistically significantly (p<0.001) decreased by 58.4% relative to the control animals. CEP under the prophylactic regimen showed more pronounced anti-ulcer activity than esomeprazole, which was 96.4% and 69.2%, respectively.

In-vitro Anti-Ulcer Activities of Mallotus Japonicus

Objectives: This study's objective was to investigate whether or not a methanolic extract of Mallotus japonicas could decrease H+-K+ ATPase activity and neutralise acid. Materials and Methods: We assessed the total phenolic and flavonoid contents of the sample while it was exposed to varying amounts of standard esmoprazole and methanol extract. Results: The proton pump inhibitory activity of the extract from stomach mucosal homogenate was found to be significant (*P<0.05) and on par with the standard. Conclusions: Based on these findings, it may be concluded that the proton pump can be effectively blocked by the methanolic extract.

Exploring ex vivo peptideolysis of thymopentin and lipid-based nanocarriers towards oral formulations

The oral delivery of medicines is the most popular route of administration for patients. However, thymopentin (TP5) is only available in the market in forms for parenteral administration. In large part, this is because of extensive peptidolytic degradation in the gastrointestinal tract (GIT), which decreases the amount of TP5 available for absorption. This study aims to understand the extent of TP5 peptideolysis and determine effective inhibitors and suitable lipid-based nanocarriers to aid in the development of an effective oral delivery formulation. Enzymatic degradation kinetics of TP5 was investigated in the presence or absence of mucosal and luminal components extracted from various parts of the rat intestine, including the duodenum, jejunum, ileum, and colon. Inhibition of TP5 enzymatic peptidolysis was screened in the presence or absence of EDTA, trypsin and chymotrypsin inhibitors from soybean (SBTCI), and bestatin. TP5 with SBTCI was loaded into lipid-based nanocarriers, including microemulsions, niosomes and solid lipid nanoparticles. These TP5-loaded nanocarriers were investigated through characterization of morphology, particle size, zeta potential, entrapment efficacy (EE%), and ex vivo rat intestinal degradation studies to select a lead formulation for a future oral drug delivery study. The degradation kinetics of TP5 followed pseudo-first-order kinetics, and the biological metabolism of TP5 was displayed in the presence of luminal contents, indicating that TP5 is sensitive to luminal enzymes. Notably, a considerable decrease in TP5 peptidolysis was found in the presence of SBTCI, bestatin, and EDTA. TP5 and SBTCI were loaded into three lipid-based delivery systems, displaying superior protection under ex vivo intestinal luminal contents and mucosal homogenates for 6 h compared with the pure drug solution. These findings suggest that using select inhibitors and lipid-based nanocarriers can decrease peptide degradation and may improve oral bioavailability of TP5 following oral administration.

Gastroprophylactic Effects of p-Cymene in Ethanol-Induced Gastric Ulcer in Rats

The prevalence of gastric ulcers has increased in recent years, mainly because of non-steroidal anti-inflammatory drug utilization. Therefore, the current study investigates the gastroprotective effect of p-Cymene on absolute ethanol-induced acute gastric mucosal hemorrhagic lesions in rats. Thirty Sprague Dawley rats were randomly separated into five groups: normal control, ulcer control, reference, and two experimental groups. The normal and ulcer control groups were orally fed with 0.5% carboxymethylcellulose (CMC). The reference group was fed orally with 20 mg/kg omeprazole. The experimental groups were fed with 30 mg/kg and 60 mg/kg p-Cymene, respectively. After one hour, the normal group was fed with 0.5% CMC, and groups 2–5 were given absolute alcohol. After another hour all rats were sacrificed. The ulcer control group showed severe superficial hemorrhagic gastric mucosal lesions with decreased gastric mucus secretion and pH of gastric content. p-Cymene significantly reduced ethanol-induced gastric lesions, as evidenced by increased mucus and pH of gastric content, decreased ulcer area, reduced or absence of edema, and leucocyte infiltration of the subcutaneous layer. In gastric mucosal homogenate, p-Cymene displayed a significant increase in superoxide dismutase (SOD), catalase (CAT) activities, prostaglandin E2 (PGE2), and significantly reduced the malondialdehyde (MDA) level. In addition, p-Cymene increased the intensity of periodic acid–Schiff (PAS) stain of the gastric epithelium, and produced up-regulation of the HSP 70 protein and down-regulation of the Bax protein of the stomach epithelium, as well as a reduction in the levels of tumor necrotic factor-alpha and interleukin-6, while the level of interleukin-10 was increased. p-Cymene decreased the level of TNF-a and IL-6, and increased the level of IL-10. Acute toxicity with a higher dose of 500 mg/kg p-Cymene did not manifest any toxicological signs in rats and could enhance defensive mechanisms against gastric mucosal lesions. p-Cymene showed gastroprotective effects that could be attributed to its antioxidant nature, or its ability to increase mucus secretion, increase endogenous enzymes (SOD, CAT, PGE2), reduce MDA level, up-regulate HSP 70 protein, down-regulate Bax protein, and modulate inflammatory cytokines.

Dietary Oligosaccharides Differentially Modulate Microbiome, Enteric Glia, and Epithelial Barrier Function in the Neonatal Jejunum and Colon

In our pig intestinal ischemia model, a neonatal defect in epithelial barrier restitution in the jejunum can be rescued by ischemic mucosal homogenate from weaned pigs. This is associated with an immature enteric glial cell (EGC) network, a known driver of restitution that matures postnatally partly in response to microbial colonization, which can be modulated with dietary prebiotic fiber. Therefore, we hypothesized that dietary oligosaccharide supplementation accelerates postnatal microbial colonization and EGC network maturation in the jejunum and colon, thus enhancing restitution after ischemic injury. After 24‐hours of colostrum, piglets were fed control or oligosaccharide‐supplemented formula (control‐fed or prebiotic‐fed) for 21 days and fecal swabs were sequenced for 16S rDNA. Intestinal samples were collected for western blot, imaging, and EGC culture. Surgically ischemia‐injured jejunal and colonic mucosal samples from select day 14 pigs were recovered ex vivo while monitoring epithelial barrier function by transepithelial electrical resistance (TEER). Migration abilities, calcium responses to ATP, paracrine effects on IPEC‐J2 cell restitution, and protein secretome were assessed in jejunal and colonic EGC cultures. Colonic microbial taxa changed in a time‐ and diet‐dependent manner with the prebiotic‐fed taxa clustering by day 7 and becoming progressively more tightly clustered over time (P<.050). TEER and histology of uninjured jejunum and colon were unaffected by diet. Following ischemia, low initial TEER recovered to control levels in the control‐fed colon but not prebiotic‐fed colon (diet and injury interaction, P=.038), while diet had no effect on jejunal TEER recovery. Prebiotic‐fed colon had lower levels of the EGC markers glial fibrillary acidic protein (GFAP) and S‐100B at day 21 (P<.050), and subjectively reduced density of GFAP+ and S‐100B+ EGC were noted in preliminary volume imaging of prebiotic‐fed jejunal submucosa at days 14 and 21. EGC from prebiotic‐fed colonic submucosa showed decreased chemotactic motility toward sterile‐filtered colonic contents (P=.010), decreased intracellular calcium response to ATP (P=.0075), and their co‐culture with IPEC‐J2 enhanced epithelial restitution versus monoculture (P=.032). Oppositely, EGC from prebiotic‐fed jejunal submucosa showed increased intracellular calcium response (P=.050) and their co‐culture with IPEC‐J2 did not enhance restitution as efficiently (P=.33) as those from control‐fed jejunal submucosa (P=.019). For future study, prebiotic‐fed colonic submucosal EGC differentially secreted 13 proteins of interest versus control‐fed. Preliminary results indicate dietary oligosaccharides in neonates exert different effects in the jejunum versus the colon on EGC network development and phenotype, and on epithelial restitution in vivo and in vitro. Ongoing work to understand microbiome‐ EGC‐epithelial interactions during postnatal development may lead to novel preventative and clinical practices to improve intestinal health in vulnerable neonates.

Immunization With Recombinant Haemonchus contortus Y75B8A.8 Partially Protects Local Crossbred Female Goats From Haemonchus contortus Infection.

Haemonchus contortus Y75B8A.8 (Hc8) derived from H. contortus excretory–secretory (ES) products was identified as a functional inhibitor of goat interleukin 2 (IL-2). It may act as a vaccine candidate for the development of therapeutic strategies against H. contortus infection. In this research, recombinant Hc8 (rHc8) and goat anti-rHc8 polyclonal antibodies were employed to evaluate the protective capacities of Hc8 antigen against H. contortus infections via active and passive immunization trials, respectively. In both trials, local crossbred female goats aged 9–12 months old were randomly divided into three groups, five in each group, respectively. Parasitological examinations, including fecal egg counts (FEC), cumulative FEC (cFEC), and worm burdens, were performed. In addition, antibody levels in mucosal homogenate (MH) samples and hematological and immunological parameters were detected. In the passive trial, goats were intravenously immunized with 5 mg total IgG containing anti-rHc8 goat polyclonal antibodies. After twice immunization, compared with the challenged control group, cFEC was reduced by 39%. In addition, there was a 46% reduction of worm burdens compared with the challenged controls. In the active immunization trials, 500 μg of recombinant Hc8 protein was given subcutaneously twice to 9–12-month-old local crossbred female goats with a 2-week interval, resulting in the generation of high levels of antigen-specific circulating antibodies. Besides, cFEC and abomasal worm burden were reduced by 70 and 55%, respectively, compared with the challenged control group. In addition, immunized goats had higher mucosal homogenate IgA and hemoglobin levels than the challenged controls in both passive and active immunization trials. These preliminary results demonstrated the immunoprophylactic effects of Hc8 antigen and will inform new studies on ES proteins in developing subunit recombinant vaccines against H. contortus.

Prophylactic Protective Action of Aqueous Extract of Green Oak Acorns on Ethanol-Induced Acute Injury to Rat Gastroduodenal Mucosa.

Quercus ilex fruit is widely used in the treatment of various gastrointestinal disorders, including diarrhea, for its bioactive compounds and astringent property. The current study focuses on the phytochemical characterization of the Q. ilex-aqueous extract (QIAE) and its protective effect against gastroduodenal (GD) ulcer (GDU) produced by absolute ethanol (EtOH) intoxication in adult male Wistar rats. Experimental rats were divided into six groups (n = 6): control, EtOH [95%, 4 g/kg body weight (b.w.)], EtOH + different doses of QIAE (100, 200, and 400 mg/kg, b.w.), and EtOH + Famotidine (FAM, 10 mg/kg, b.w.). Animals were orally pretreated (p.o.) with QIAE for 15 days and intoxicated with a single oral administration of EtOH for 2 h. The findings showed that the QIAE is rich in phenolic-astringent compounds and fibers, and it exhibited a significant scavenging activity on DPPH/ABTS free-radicals with half maximal inhibitory concentration (IC50) values of 177.00 ± 5.11 and 203.9 ± 2.23 μg/mL, respectively. In vivo part, QIAE significantly reduced the GD mucosal injury revealed by edema and leukocyte infiltration of the submucosal layer. GD mucosal homogenates revealed a remarkable increase in endogenous antioxidant enzyme activities (catalase, superoxide dismutase, and glutathione peroxidase) and a decrease in the lipid peroxidation levels (malondialdehyde) in animals pretreated with QIAE compared with the ulcer control group. QIAE exerted significant and dose-dependent anti-GDU protection in the rat model with a more effective action than FAM. The GD protective effect of the QIAE might be related to a direct radical scavenging activity, increased antioxidant enzymes, and depression of lipid peroxidation.

Gastroprotektivne karakteristike krioprezerviranog ekstrakta placente kod kombinovanog delovanja niskih temperatura i inhibicije ciklooksigenaze

Introduction. Non-steroidal anti-inflammatory drugs (NSAIDs) are ranked first among the world's effective anti-inflammatory and analgesic drugs with anticipated side effects. That is why the prevention of development of adverse reactions associated with NSAIDs, in particular, of those related to their ulcerogenicity, remains a serious global problem. Aims. To characterize the mechanisms of gastrocytoprotective activity of cryopreserved placenta in combined action of low temperatures and diclofenac sodium. Material and methods. The study was performed on 42 male rats weighing 200-220 g. Acute diclofenac sodium-induced gastropathy was reproduced by a single injection of the latter in rats at the dose of 50 mg/kg. The content of malonic dialdehyde, catalase activity, prostaglandin synthase activity and the content of nitrogen monoxide metabolites in homogenates of gastric mucosa were determined by spectrophotometric method. Results and discussion. The study showed that prophylactic administration of placental cryoextract in rats with diclofenac sodium-induced gastropathy is associated with increased activity of antioxidant system in gastric mucosa as demonstrated by an elevated catalase activity by 40.0% as compared with control rats. Modulation of antioxidant-prooxidant homeostasis is believed to be one of the principal mechanisms of gastrocytoprotective action in combined action of low temperatures and cryoextract of the placenta. This is shown by a statistically significant (p < 0.05) 2.2-fold increase of antioxidant-prooxidant index in the study group as compared with rats with diclofenac sodium-induced gastropathy. Administration of placental cryoextract was found to increase prostaglandin synthase activity in rats with diclofenac sodium-induced gastropathy by two times as compared with control rats, which would reduce iatrogenic prostaglandin deficiency in gastric mucosa. Also, the combined action of low temperatures and of placenta cryoextract was associated with a statistically significant (p < 0.05) increase in the level of metabolites of nitrogen monoxide (by 70.1%) as compared with rats with diclofenac sodium-induced gastropathy. Conclusions. Modulation of prooxidant-antioxidant homeostasis in gastric mucosa and increase in contents of nitrogen monoxide metabolites and prostaglandin synthase activity are the leading mechanisms of gastroprotective activity of cryopreserved placental extract in diclofenac sodium-induced gastropathy.

CHARACTERISTICS OF THE MECHANISMS OF CYTOPROTECTIVE ACTIVITY OF CRYOPRESERVED PLACENTA EXTRACT ACCORDING TO MORPHOLOGICAL AND BIOCHEMICAL STUDIES OF THE GASTRIC MUCOSA IN THE EXPERIMENT

Relevance. Non-steroidal anti-inflammatory drugs (NSAIDs) are used as antipyretics and analgesics. In this case, NSAIDs have a side effect - ulcerogenic. Therefore, it is important to find means to correct the ulcerogenic action of NSAIDs. Potential antiulcerogenic action may be in cryopreserved human placenta extract "Cryocell-placental cryoextract".
 Objective: to investigate the effect of cryopreserved placenta (CEP) extract on the morphological state of the stomach and the processes of protein synthesis in the gastric mucosa with the introduction of NSAIDs diclofenac sodium in rats with experimental rheumatoid arthritis.
 Materials and methods. The study was performed on 28 nonlinear male rats weighing 200-220 g. The animals were divided into 4 groups: I (n = 7) – intact rats; II (n = 7) – rats with adjuvant arthritis (AA), this is a treatment without treatment; III (n = 7) – rats with AA, treated with diclofenac sodium (DN), 8.0 mg / kg, intragastrically (i/g); IV (n = 7) – rats with AA, treated with DN, 8.0 mg / kg, i/g and CEP, 0.16 ml / kg body weight, intramuscularly (i/m).
 AA was simulated by subplantar injection of complete Freund's adjuvant into the hind right limb at a rate of 0.1 ml per rat. The day of adjuvant administration was considered as the "0" day of the experiment. AA treatment was performed from 14 to 28 days. DN was applied daily, and CEP – with an interval of 3 days: for 14, 17, 20, 23 and 26 days. On day 28, the animals were removed from the experiment by cervical dislocation under inhalation anesthesia.
 To assess morphological changes, gastric fragments were fixed in 10.0% neutral formalin solution. Histological sections 5–7 μm thick were stained with hematoxylin and eosin, as well as Schiff's reagent (fuchsin sulfuric acid) with iodic acid. The concentration of total protein (TP) was determined spectrophotometrically by light absorption at a wavelength of 546 nm. The content of RNA and DNA in the homogenates of the gastric mucosa was determined spectrophotometrically by reaction with perchloric acid.
 Results. In rats with AA, the mucous membrane (MM) had a preserved structure in all animals. There were no signs of hyper- or hyposecretion of mucins when stained with a chic reaction with alcyan blue. There were foci of desquamation of the superficial epithelium and single lymphocytes. The introduction of DN in rats with AA led to signs of erosive changes in MM. Deep erosions and single ulcerative defects were found in 28.6% of rats, reaching the depth of the lesion to the muscle plate. There is a small amount of mucopolysaccharides with a predominance of acidic glycoproteins. Reduced content of neutral glycoproteins. Single desquamated epitheliocytes were detected. Focal lympho-plasmacytic infiltration with exudate with eosinophilic leukocytes was noted in the submucosal layer of MM. Against the background of the combined use of DN and CEP in rats with AA in any animal, no significant violations of the histoarchitectonics of the MM, except for isolated foci of desquamation of the superficial epithelium.
 In gastric MM homogenates in rats with AA there was a decrease (p <0.01) in the level of TP by 14.1% and a decrease (p <0.05) in the content of RNA and DNA by 27.5% and 14.3%, respectively, relative to indicators of intact animals.
 Against the background of DN monotherapy, there was an even greater decrease (p <0.001) in the level of TP: by 26.2% (40.3 ± 1.43 μg / mg) relative to intact animals (54.6 ± 1.39 μg / mg). And the content of RNA and DNA was 0.59 ± 0.57 μg / ml and 2.21 ± 0.27 μg / ml, respectively.
 Against the background of the combined use of DN and CEP, comparable to the level of intact animals indicators of TP and DNA were noted. Thus, the level of TP was 53.0 ± 2.09 μg / mg, which was only 2.9% lower than intact animals (54.6 ± 1.39 μg / mg), and the DNA content by 5.1% (p = 0.3) was lower than intact animals (3.1 ± 0.12 μg / ml) and was respectively 2.94 ± 0.08 RNA level (1.72 ± 0.12 μg / ml) on 17.6% exceeded (p = 0.1) intact animals (1.46 ± 0.08 μg / ml).
 Conclusions. The combined use of placental cryoextract and diclofenac sodium eliminates the negative effect of the latter on protein metabolism in the gastric mucosa of rats with adjuvant arthritis. Thus, the level of total protein was 53.0 ± 2.09 μg / mg, which is only 2.9% lower than intact animals. Restoration of diclofenac sodium-induced mucin secretion by the integumentary epithelium of the gastric mucosa is one of the mechanisms of cytoprotective activity of cryopreserved placenta extract in experimental ulcerogenesis.

Possible Regulation of P-glycoprotein Function by Adrenergic Agonists in a Vascular-luminal Perfused Preparation of Small Intestine

Although the functions of small intestine are largely regulated by enteric nervous system (ENS), an independent intrinsic innervation, as well as central nervous system (CNS), the neural regulation of drug absorption from the small intestine still remains to be clarified. To obtain some information on it, the effect of adrenergic agonists on P-glycoprotein (P-gp) function was investigated by utilizing a vascular-luminal perfused rat small intestine. Adrenaline significantly decreased the secretion of rhodamine-123 (R-123) into the intestinal lumen, but dibutyryl cAMP (DBcAMP) significantly enhanced R-123 secretion. The inhibition study with quinidine clearly indicated that the decrease in secretory clearance of R-123 by adrenaline or the increase by DBcAMP would be attributed to the decrease or increase in P-gp activity, respectively. Expression levels of P-gp in whole mucosal homogenates were not changed at all by any chemicals examined, but those on brush border membrane (BBM) of intestinal epithelial cells were significantly decreased or increased by adrenaline or DBcAMP, respectively. Furthermore, changes in P-gp activity caused by adrenergic agonists and DBcAMP were significantly correlated with changes in expression level of P-gp in BBM, suggesting that the trafficking of P-gp from cytosolic pool to BBM would be regulated by adrenergic agonists and DBcAMP.

BACTERIAL SYMBIOSIS IN COMPLICATED ULCERS: THE PATHOGENETIC HYPOTHESIS

Aim of this paper is to estimate crystallogenic properties of gastric mucosa in connection with its microbial contamination. We investigated crystallogenic properties of some biological substrata (gastric mucosa, gastric mucosal layer homogenates) in 12 healthy individuals and 30 patients with ulcer disease complicated in 12 cases by perforation, bleeding or penetration. Biological substrata were received at fibrogastroduodenoscopy. Estimation of crystallogenic properties of biological material was accomplished by classic crystalloscopy. Biological substrata crystalloscopic investigation was accompanied by its traditional microbiological study for Helicobacter pylori and detection of other microorganisms. Our data allow to suppose dual contamination of stomach mucosa both by Helicobacter pylori and Providencia or Morganella. This combination caused elevation of gastric mucosa crystallogenic properties that provoked formation of ulcer. Procrystallogenic potential of this symbiosis may be an important link to the pathogenesis of ulcer disease which realized through microorganism-associated mucosal damage and the progression of complications.

Maternal Supplementation of Clofibrate Stimulates Intestinal Fatty Acid Oxidation in Newborn Suckling Piglets

Abstract Objectives To evaluate the effects of maternal feeding of clofibrate, a pharmaceutical PPARα agonist, on the development of intestinal fatty acid metabolism in offspring using the pig as a model. Methods A total of 27 pregnant sows were randomly assigned into three treatment groups. Each group was fed a standard diet (3265 kcal ME/kg) supplemented with either 0, 0.25% or 0.5% clofibrate (w/w) from d 107 of gestation to d 7 of lactation. A piglet from each litter was sampled on 1, 7, 14 and 19 d of age. Fatty acid oxidation was examined in homogenates of ileum mucosa using 1 mM [1–14C] oleic acid as a substrate in the absence or presence of in vitro supplemented L-carnitine (1 mM), malonate (10 mM) or carnitine + malonate. Results The metabolism of [1–14C] oleic acid was modified by the maternal clofibrate, and the effects varied with the dose of clofibrate and postnatal age (p < 0.001). The 14C accumulation in 14CO2 increased by 2.4 fold at d7 (vs d1), but showed no difference after d7. Maternal clofibrate stimulated the 14CO2 accumulation from d1 to d19, but the stimulation decreased with postnatal age. The 14C accumulation in acid soluble products (14C-ASP) was reduced by 8.4 fold at d7 (vs d1), and no difference was observed after d7. Maternal clofibrate increased 14C-ASP at d1, and the effect was greater as maternal clofibrate was incremented from 0.25 to 0.5%. The 14C accumulation in esterified products (14C-ESP) was greater at d7 than d1, 14 and 19. Maternal clofibrate reduced the 14C-ESP and, again the reduction was stronger with increasing clofibrate dose. In vitro supplementation of carnitine increased 14CO2 by 60% and 14C-ASP by 120%, but reduced 14C-ESP by 39% compared to control incubations. Supplementation of malonate reduced 14CO2 by 95% and 14C-ESP by 44%, but no effect on 14C-ASP was detected. Conclusions Maternal dietary supplementation of clofibrate increases intestinal fatty acid oxidation in suckling offspring, but the increase is attenuated after one week. Inhibition of the TCA cycle reduces CO2 production without affecting ASP. The availability of carnitine could be a key element to maintain a high rate of intestinal fatty acid oxidation during the neonatal period. Funding Sources Supported by Program of Animal Nutrition, Growth and Lactation [grant no. 2015–67015-23245/project accession no. 1005855] from the USDA NIFA.

IN VITRO ANTI-ULCER ACTIVITY OF DIFFERENT EXTRACTS OF CISSUS QUADRANGULARIS

Aim:The aim of this study was undertaken to assessin vitro H+-K+ ATPase inhibitory activity of different extracts of Cissus quadrangularis Linn. Materials and Methods: Phytochemical and Phyto-analytical studies like total phenolic compound and total flavonoid contents from extract were quantified and H+-K+ ATPase inhibition assay was performed in presence of different concentrations(10,20,50 and 100 μg/ml) of standard (omeprazole),chloroform and methanolic extract. Results:The extract has shown dose dependent significant (*P < 0.05) proton pump inhibitory activity in the goat gastric mucosal homogenate which was compared to standard Omeprazole. Conclusions: Hence, from this study we have concluded that methanolic extract of C.quadrangularis has more inhibitory effect than chloroform extract.