Histological and Immunohistochemical Investigation Concerning the Potential Protective Function of Quercetin and Omega-3 against the Detrimental Effects of Energy Drinks on The Pancreas of Adult Male Albino Rats

Background: Pancreas is one of the most vital complex organs in the body. Bleomycin sulfate is a cytotoxic drug that may injure the pancreas. Objective: To detect the effect of bleomycin sulfate on pancreas of adult male Albino rats and to assess the possible pancreatic protection by selenium use. Materials and methods: This study continued 10 days and included 20 adult male Albino rats that were categorized into three groups: group I served as the control group. Group II: bleomycin sulfate treated group. Group III: bleomycin sulfate and selenium treated group. Pancreatic specimens were excised and processed for light and electron microscopic examination. Furthermore, Anti-insulin immunohistochemical staini was assessed and statistically analyzed. Results: Treatment with bleomycin sulfate induced distortion and damage of pancreatic architecture with areas of hemorrhage and necrosis. The acinar cells showed vacuolation of cytoplasm with either euchromatic or pyknotic nuclei. Islets of Langerhans appeared distorted, shrunken with difficult differentiation between beta cells and alpha cells. Most of these changes were improved by selenium administration. Conclusion: Selenium has significantly ameliorated bleomycininduced alterations in the pancreas of adult male Albino rats.

Background: The world wide increasing popularity of the energy drinks and the lack of information about their possible hazardous effects on health is a matter of controversy and research. The aim of this study is to assess the histological and histochemical effects of energy drinks on the pancreas of adult male albino rats and the possible protective effect of omega-3. Material and Methods: Fifty adult male albino rats were randomly divided into 4 groups. First group is control. Second group (Omega-3 treated) rats received omega-3 at a dose of 300 mg /kg/day orally for 4 weeks. Third group (Red Bull treated) rats received Red Bull at a dose of 10 mg/kg/day orally for 4 weeks then they were randomly subdivided into two equal subgroups: IIIA, rats were sacrificed after 24 h of the last dose and in IIIB (Recovery group), rats were sacrificed after 4 weeks of the last dose .Fourth group (Omega-3 and Red Bull treated group) rats received Red Bull at a dose of 10mg/kg/day and Omega-3 at a dose of 300mg/kg/day for 4 weeks. At sacrifice, blood samples were drawn for biochemical study and pancreas specimens were prepared for histological and histochemical study. Results: Energy drink had no significant effect on the animal weight (P = 0.055), but there was highly significant increase in the pancreatic weight (P = 0.001) and in mean blood glucose level (P = 0.000). There were signs of β cells overstimulation. Histological and histochemical study of the pancreatic sections revealed multiple deleterious effects of the energy drink on the acinar and the islet cells. These changes were reversible as shown in the recovery group. Co-administration of Omega-3 showed marked protection of the pancreatic acini and the islets of Langerhans. Conclusion: Omega-3 administration has a highly protective effect on the pancreatic tissue against the hazardous effects of the energy drinks.

Bisphenol A (BPA) is an endocrine disturbing element liberated through the environment and broadly used all over the world. The purpose of this work was to explore the impacts of BPA on liver, spleen and pancreas of adult male albino rats and the probable protecting role of Lycopene. Thirty adult male albino rats were distributed into three groups; group I (control group) , group II administrated BPA orally (50 mg/kg) for 30 days and group III (BPA+ Lycopene treated group) administered Lycopene (LYC) concomitantly with BPA at a dose of 10 mg/kg. At the end of the experiment, rats of all groups were sacrificed. Liver, spleen and pancreas were extracted, fixed and processed for histologic study. The area % of the collagen fibers in liver, spleen and pancreas was measured in the different groups and statistically analyzed. Structural alterations were discovered in liver, spleen and pancreas of BPA-treated rats including dilated congested blood vessels and vacuolar degeneration of the cells. When Lycopene (LYC) was concomitantly administered with BPA, It produced marked improvement at structural and ultrastructural levels. The morphometric results declared that the area % of the collagen fibers in the liver, spleen and pancreases was considerably increased in the BPA-treated group in comparison to the control group. Meanwhile the area % of the collagen fibers of BPA+ Lycopene treated group was more or less comparable to the control group with no significant difference. It was concluded that LYC has noticeable protective effects versus the damaging effects of BPA.

Introduction: Energy drinks are widely used among youth as boosters to increase their concentration. Many Health disorders associated with energy drinks have been reported. Avocado oil is a natural oil extracted from avocado fruit and possess an antioxidant and anti-inflammatory effects. Aim of the work: This work was conducted to investigate the effect of energy drink on the histological structure of the pancreas of adult male albino rat and to evaluate the possible protective role of avocado oil.Materials and Methods: Forty adult male albino rats were divided equally into 4 groups; control group, avocado oil-treated group (0.4 mg/100 gm body weight), energy drink-treated group (1.5 mg/100 gm body weight) and both avocado oil and energy drink treated group. All animals were orally administrated once daily for 4 weeks. Pancreatic specimens were processed for light and electron microscopy. Immunohistochemical study was performed using insulin and caspase-3 antibodies. Results: Energy drink-treated group revealed focal disturbed architecture of the exocrine and endocrine parts of pancreas. The exocrine pancreas appeared with cytoplasmic vacuolations and deeply stained nuclei of the acinar cells in addition to congested blood vessels. Ultrastructural examination revealed both acinar and islet cells with cytoplasmic vacuolations, swollen mitochondria with disrupted cristae, dilated rough endoplasmic reticulum in addition to shrunken condensed nuclei with dilated perinuclear cisternae. The immunohistochemical study showed a significant decrease in the insulin immunoreaction of β cells associated with a significant increase in caspase-3 immunoreaction in acinar and islet cells. In contrast, minimal changes were observed in rats treated concomitantly with avocado oil and energy drink with a non-significant change in the immunohistochemical reaction. Conclusion: Energy drink induced structural changes in the pancreas of rats and the concomitant intake of avocado oil could to some extent ameliorate such changes

Depression is linked with a high risk of type 2diabetes (T2D). The affiliation between depression and diabetes might be correlated to depression itself and, or medications recommended. Significantly, the usage of selective serotonin reuptake inhibitors (SSRIs), the most commonly antidepressants increased the hazard of developing T2D. Nevertheless, the mechanism underlying this suggestion remains vague. Omega-3 had antioxidant and anti-inflammatory activity. So, there is developing evidence that consumption of omega-3 could be expedient. The present study was carried out to investigate the potential effect of omega-3 in the fluoxetine induced alterations in the pancreas of adult male albino rats. Forty adult male albino rats were divided into four groups. Group I served as a control group. Group II received a single daily dose of 300 mg/kg of omega-3. Group III received 24 mg/kg bw/day of fluoxetine hydrochloride. Group IV received omega-3 and fluoxetine as group II and III for 30 days. Light and electron microscopic investigations were carried out. Histological examination using H & E and Masson’s Trichrome stain were carried out. The insulin expression in β cells was evaluated using immunohistochemistry. Morphometric results were subjected to statistical analysis. Investigation of group III (Fluoxetine group) showed distorted exocrine pancreas with thick interlobular septa that contained dilated congested blood vessels, cellular infiltration, and fat cells. Marked shrunken of the pancreatic islets was observed. Masson’s trichrome stain showed increased collagen fibers deposition. Electron microscopic examination revealed that most of the acinar cells had irregular shaped nuclei with peripheral heterochromatin and wide capillaries. The cytoplasm of β cells had a variety of secretory granules. Most of them had an electron dense core with increased electron lucent halo however, few β granules were empty and coalesced. Omega- 3 supplementation improved the morphology of Langerhans compared to fluoxetine group. Importantly, some pancreatic duct cells revealed a positive reaction against anti-insulin antibodies. The current results demonstrated that fluoxetine harmfully affected the histological structure of the pancreas. Omega-3 diminished effectively some histological, immunohistochemical and electron microscopic changes in a fluoxetine induced pancreatic injury. Omega-3 could stimulates β-cell regeneration from potent islet progenitor cells present in the ductal cells and these might lead to repair of the functional accomplishments of the injured pancreas to a great extent.

Introduction Aflatoxin contamination of foods is a worldwide problem, especially in developing countries. Ginger has antioxidant properties. Aim of the study To study the histological and biochemical changes in the pancreas of rats with experimental aflatoxicosis, and to evaluate the role of ginger supplementation. Materials and methods Forty-five adult male albino rats were equally divided into three groups. Group I that served as the control group. Group II that received 250 μg/kg body weight/day of aflatoxin B1 dissolved in olive oil using a gastric tube for 5 days/week for 4 weeks. Group III that received both aflatoxin as in group II and 400 mg/kg body weight/day of ginger orally for 5 days/week for 4 weeks. At the end of the experiment, all rats were anesthetized, and their pancreases were extirpated and divided into two parts to be processed for light and electron microscopic examinations. Morphometrical analysis for area percentage of collagen fibers and biochemical analysis for glucose, insulin, and serum amylase were performed and statistically analyzed. Results Examination of group II revealed thick interlobular septa that contained congested blood vessels, cellular infiltration, mast, and fat cells. Pancreatic acinar cells showed decreased secretory granules, vacuolization, and dilated fragmented rough endoplasmic reticulum. Few acinar cells showed rarified areas of cytoplasm. Some acinar cells had small condensed heterochromatic nuclei. Most of the islets of Langerhans were formed of cells separated by dilated congested capillaries. Most of the nuclei of β cells were euchromatic, whereas some were small heterochromatic. The cytoplasm of β cells had a variety of secretory granules. Most of them had an electron-dense core and an electron-lucent halo, whereas others had homogenous moderate density. Some granules coalesced. A few cells had cytoplasmic areas depleted of granules. Pancreatic ducts were dilated. Examination of group III revealed that pancreatic lobules were separated by thin interlobular septa. Acini had numerous apical acidophilic secretory granules, a few vacuoles, and basal euchromatic nuclei. Beta cells of the islets of Langerhans had euchromatic nuclei and numerous secretory granules with an electron-dense core and a wide electron-lucent halo. Biochemical analysis of glucose and serum amylase showed a highly significant increase, whereas that of insulin showed a highly significant decrease, in group II in comparison with group I. The glucose and serum amylase levels were significantly decreased, whereas the insulin level was significantly increased in group III compared with group II. Conclusion Aflatoxin had a deleterious effect on the histological structure of the rats' pancreas, and ginger minimized these effects.

Biochemical changes associated with the administration of aqueous pulp extract of Irvingia gabonensis fruit on adult male Wistar rats

Background Ethanol has a destructive effect on the pancreas by inducing oxidative damage on pancreatic cells. This injurious effect could be ameliorated by the administration of vitamin E. Aim The aim of the study was to investigate the possible protective role of vitamin E on ethanol-induced alterations in the pancreas in adult male albino rats and also the effect of ethanol withdrawal. Materials and methods Forty adult male albino rats were divided into four equal groups (10 rats each): group I (control group), group II (ethanol-treated group), group III (ethanol and vitamin E-treated group), and group IV (recovery group). At the end of the experiment, the rats were sacrificed by cervical dislocation and their pancreas were excised and prepared for light microscopic and histochemical study. Detection of interleukin (IL)-1B and IL-6 by real-time PCR and proliferating cell nuclear antigen immunoexpression in the pancreatic tissue was carried out. Results Group II showed loss of acinar tissue, distorted degenerated pancreatic acini, dilated distorted interlobular ducts, distorted islets of Langerhans, and dilatation and congestion of the blood vessels. Interlobular inflammatory cellular infiltration was also detected. There was an increased collagen fiber deposition and diminution of the proliferating stem cells. There was a marked increase in IL-1B and IL-6 levels. Group III showed an obvious improvement in the structure of the pancreas. The recovery group showed improvement in the structure of the pancreas and minimal collagen deposition. Conclusion Vitamin E could protect the pancreas against ethanol-induced alteration in adult male albino rats.

Objectives The aim of this study was to investigate the effect of furan on the pancreas of adult male rat and to evaluate the possible protective role of garlic oil (GO). Background Furan is carcinogenic in rats and mice and possibly carcinogenic to humans. Methods Sprague–Dawley rats were divided randomly into four main groups: control, GO (80 mg/kg/day), furan-treated (2 and 8 mg/kg/day), and protected group (furan and GO). All rats were treated orally by a gavage for 5 days per week for 8 weeks. At the end of the experiment, pancreases were subjected to biochemical (measurement of the level of glucose, amylase, lipase, and oxidative stress indices in pancreatic tissue), histological, and immunohistochemical analyses. Results Our results showed the toxic effects of furan on the pancreas in adult male albino rats. This was indicated by an increase in pancreatic (amylase and lipase levels) and lipid peroxidation biomarker (malondialdehyde), and a decrease in antioxidant enzymes, including catalase, superoxide dismutase, and glutathione. In addition, histopathological alterations were detected, including mononuclear cellular infiltration, congestion of blood vessels, and cytoplasmic vacuolation of the acinar cells. There were significant increases in the number of inflammatory cells and apoptotic expression, whereas cytochrome P450 2E1 expression was significantly decreased after the administration of furan. Furan-induced toxicity was ameliorated by the coadministration of GO. Conclusion The administration of furan-induced biochemical and histopathological changes in the pancreas of adult male albino rats. These changes were improved by the coadministration of GO.

In fetal rats, just before birth, there is a dramatic increase in pancreatic amylase activity. Besides the well demonstrated role of corticosteroids in this process, in vivo studies have suggested that CCK, a trophic hormone for the pancreas in adult rats, might also be involved (Werlin, Biol Neonate 1983; 44: 287-94). Since in vivo studies cannot rule out an indirect maternal effect through corticosteroid secretion, we studied the direct effect of CCK on fetal rat pancreas in organ culture. Furthermore, the role of insulin, another important hormone for the exocrine pancreas in adult rats, was checked by using streptozotocin (STZ). Pancreata from 20 day-old rat fetuses were cultured in a serum-free medium (SFM) for 6 days, with or without dexamethasone 3.10−6M (DXM) and/ or CCK8 2.10−6M. In the presence of these two hormones, and with or without insulin (0.1 U/ml), cultures were exposed to STZ 10−7M for the first day of culture. Pancreatic explants were assayed for proteins and amylase on days 0, 2, 4, 6. Amylase specific activity (ASA) was expressed as percentage of day 0. With SFM alone, almost all the ASA disappeared by day 2 (17.7 % ± 12.5), but was partially but significantly maintained when CCK (48.6% ± 8.8) or DXM (47.9% ±14.4) was added. The DXM effect was significantly maintained for 6 days but not the CCK effect which lasted only 2 days. On day 2 of culture the latter was dose-dependent with a maximum occuring between 10 md 10−10M and was inhibited by asperlicin 10 μM. A combination of CCK and DXM gave the best results in maintenance of ASA (87.4 % ± 21.8 on day 2; 81.9% ± 15.9 on day 4; 62.2% ± 10.3 on day 6). When cultures in this optimal medium (CCK + DXM) were exposed to streptozotozin for the first day of culture, a significant decrease of the ASA was found on days 4 (63.2% ± 21)and 6 {47.6% ± 12.8). This difference was corrected when, in the same protocol, insulin was added for the complete time of culture. Conclusions: CCK, like DXM, is important in the prenatal development of the pancreas in the rat. Its action on ASA is different from and additive, but not synergistic, to that of DXM. The effect of STZ, corrected by insulin, suggests also a role for this hormone. The paracrine effect of insulin on exocrine pancreas, well demonstrated in the adult rat, might be already efficient in the fetus.

Aim of the Study: G-CSF and umbelliferon were tested to see if they could reduce olanzapine-induced pancreatic damage in adult male albino rats. Olanzapine's pancreatic damage and G-CSF and umbelliferon's protective effects were examined in terms of histological, immunohistochemical, and biochemical changes. Study Type: A randomised, controlled study examined the effects of olanzapine, G-CSF, and umbelliferon on pancreatic tissue. Study place: Rashid Latif Medical College, Lahore. Research Method: 40 mature male albino rats were divided into four ten-rat groups: Regular saline was provided to the Control Group for eight weeks. The Olanzapine Group received 10 mg/kg/day oral olanzapine for eight weeks. The Olanzapine + G-CSF Group received 10 mg/kg/day intraperitoneal olanzapine and 50 µg/kg/day G-CSF for 8 weeks. Oral olanzapine (10 mg/kg/day) and umbelliferon (50 mg/kg/day) were given to Olanzapine + Umbelliferon Group participants for eight weeks. After the experiment, blood and pancreatic tissues were collected for histological, immunohistochemical, and biochemical investigation. The histological study included H&E and Masson's trichrome staining. The immunohistochemistry examination assessed the expression of caspase-3, TNF-α, and insulin. Both MDA and SOD, as well as GPx and metabolic parameters (fasting glucose and insulin), were investigated by the use of biochemical assays. The data were examined with SPSS 25 using a one-way analysis of variance (ANOVA) and Tukey's post hoc test. Results: 1 The olanzapine group demonstrated substantial pancreatic architectural deformation, collagen deposition, and cellular infiltration, according to the histological findings. Umbelliferon and G-CSF work synergistically to improve pancreatic architecture while concurrently lowering collagen. According to the immunohistochemistry data, the group that was provided olanzapine exhibited heightened levels of apoptosis and inflammation, as shown by the increased expression of Caspase-3 and TNF-α. A reduction in insulin expression indicates β-cell malfunction. Umbelliferon and G-CSF restored insulin levels and significantly reduced caspase-3 and TNF-α expression. 3 Biochemical Results: Oxidative stress was shown by increased MDA and decreased SOD and GPx activity in the olanzapine group. Fasting glucose increased while insulin declined. The treatment of G-CSF and umbelliferon normalised metabolic and oxidative stress indicators. Discussion: The study showed that olanzapine causes pancreatic oxidative stress, inflammation, and apoptosis. Umbelliferon and G-CSF prevented these pathologies. G-CSF and umbelliferon, due to their tissue-repairing and anti-inflammatory properties, improved pancreatic architecture, reduced oxidative stress, and increased β-cell function. These findings suggest G-CSF and umbelliferon may cure olanzapine-induced pancreatic dysfunction. Conclusion: Histology, immunohistochemistry, and biochemistry show that olanzapine causes pancreatic oxidative stress, apoptosis, and inflammation. G-CSF and umbelliferon protected against olanzapine-induced pancreatic damage, suggesting they may minimise its metabolic adverse effects. These findings improve our understanding of olanzapine-induced pancreatic damage and prepare researchers for future therapeutic intervention studies..

Introduction Type I diabetes is characterized by the deficiency of endocrine β cells in the pancreatic islets of Langerhans. Stem cell therapy can be an effective therapeutic approach. Aim of the work The present study was carried out to investigate the therapeutic effect of mesenchymal stem cells (MSCs) in the recovery of streptozotocin (STZ)-induced diabetes mellitus in the pancreas of adult male albino rats. Materials and methods Thirty-five adult male albino rats were divided into three groups. Group I served as the control group. In group II, diabetes was induced by a single intraperitoneal injection of STZ at a dose of 60 mg/kg body weight. In Group III, diabetes was induced; the rats then received MSC therapy and were sacrificed after 1 week (subgroup IIIa) and at 4 weeks (subgroup IIIb). The mean blood glucose levels were measured for all groups. Pancreatic sections were subjected to the following staining procedures: H&E, MSC marker CD44, and insulin immunostaining. The mean number of CD44-immunopositive cells and the mean area % of insulin-immunopositive cells were measured using image analysis. Results were statistically compared. Results In group II, cells located mainly in the central part of the islets showed marked swelling and a vacuolated and degenerated cytoplasm. Peripheral islet infiltration by lymphocytes was observed. Treatment of the diabetic group with MSCs for the longer duration (4 weeks) caused restoration of the normal pancreatic architecture. There was a significant increase in the number of CD44 cells after MSC therapy compared with the control and diabetic groups. Mean area % of insulin-positive cells presented a significant decrease in the diabetic group, followed by progressive increase after MSC therapy. Conclusion Treatment with MSCs promoted pancreatic islet regeneration in STZ-induced diabetic rats, suggesting its potential use as a therapeutic modality for diabetes mellitus.

Background: Valproic acid [VA] is an antiepileptic drug that is extensively prescribed for many neurological disorders. Its use is associated with serious pancreatic complications. Marjoram oil [MO] is a health protective agent with proved anti-oxidant activity.Aim: Study of the effect of valproic acid on pancreas of adult male albino rats exploring novel mechanisms of VA-induced pancreatic damage and to evaluate the potential protective role of marjoram oil. Materials and Methods: Fifty adult male albino rats were used as a control group, a valproic acid group and a valproic acid-marjoram group. Both VA [200mg/kg] and MO [0.5ml/kg] were given once a day orally for eight weeks. Specimens of the pancreas were processed for light and electron microscopic studies. Immunohistochemical study was performed using anti-P53 antibodies. Results: Specimens of the valproic acid group showed an obvious distortion in the pancreatic acini as well as the islets of Langerhans. The acinar and islets cells showed cytoplasmic vacuoles and pyknotic nuclei. Large autophagic vacuoles containing an acidophilic material appeared in many cells. Dilated ducts and blood vessels were seen. There was a significant increase in the P53-immunoreaction of the acinar and islets cells and a significant decrease in the number of the zymogen granules of the acinar cells. Ultrastructurally, there were shrunken nuclei with irregularity, dilated rough endoplasmic reticulum and swollen mitochondria in the acinar cells and beta cells. In contrast, minimal changes occurred in the valproic acid-marjoram group that received MO before VA. Conclusion: Valproic acid administration to albino rats resulted in significant structural alterations in the pancreas. Marjoram oil attenuated VA effect and preserved the pancreas structure.

The effect of tetracycline, at two doses of 50 and 200 mg kg(-1) daily, was studied on pancreatic and liver tissue function for 14 and 21 days in adult male albino rats. For pancreatic function the parameters studied were content of amylase and lipase in pancreas, serum amylase and lipase, serum glucose and faecal fat excretion. For liver function, liver specific enzymes in serum, namely alanine amino transaminase, aspartate amino transaminase and lactate dehydrogenase were estimated. In addition, total lipid, antiperoxidative enzymes and lipid peroxidation were measured in pancreas and liver. The content of amylase and lipase in pancreas showed a small but significant decrease in the rats given 50 mg kg(-1) for 21 days and the decrease was much more significant in those receiving the 200 mg kg(-1) dose. In pancreas free radical levels show a significant increase and reduced glutathione shows a substantial decrease at the 50 mg kg(-1) level and a significant change in these parameters was observed at the 200 mg kg(-1) dose. Antioxidant enzymes, superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase, showed a small but significant decrease in the pancreas of the rats treated with 50 mg kg(-1) tetracycline. A significant decrease in the antioxidant enzymes level was observed at the 200 mg kg(-1) dose. In the liver, free radical levels and reduced glutathione were within the normal range at the 50 mg kg(-1) level and significant changes were observed at 200 mg kg(-1). The antioxidant status was unaffected in liver after treatment with tetracycline at the 50 mg kg(-1) level and a significant decrease was observed at the higher dose. Our results reveal the safe nature of tetracycline with respect to the liver at the lower dose tested, whereas, both the higher and lower doses seem to have detrimental effect on the pancreas as revealed by the rise in free radical levels and decrease in the antioxidant enzyme levels.

Background: Bleomycin sulfate is a chemotherapeuticantibiotic that may has adverse effects on both the exocrineand endocrine component of the pancreas.Aim of Study: This study was designed to assess the effect of bleomycinsulfate on pancreas of adult male albino rats and the possible protection by vitamin E administration.Material and Methods: Twenty-five adult male albinorats were divided into three groups: Group I: Control group,Group II: Bleomycin sulfate treated group and Group III:Bleomycin sulfate and vitamin E treated group. At the end ofthe experiment, pancreas was taken out and processed forlight, electron microscopic examinations and anti-insulinimmunohistochemical staining. The percentage area of anti-insulin antibody reaction was measured and statisticallyanalyzed.Results: Treatment with bleomycin sulfate showed variablemarked histopathological changes. Distortion of the normalarchitecture of the pancreatic tissue with areas of hemorrhage,vacuolation and necrosis either in the acini or islets of Lang-erhans with loss of differentiation between beta cells andalpha cells. Histopathological changes were diminished afterreceiving vitamin E. The acinar cells were more regular withbasophilic rounded basal nuclei and apical acidophilic cyto-plasm. The islet of Langerhans restored its normal shape andsize with nearly regular out line.Conclusion: Vitamin E could protect the pancreas againstbleomycin sulfate-induced alteration in adult male albino rats.