Investigating the Effect of Sertraline on Pancreatic Injury in Male Rats

Objective To investigate the correlation between pancreatic nuclear factor-κB (NF-κB) activation, cell apoptosis and pancreatic injury. To determine effects of N-acetylcysteine (NAC) on pancreatic injury in rats with acute necrotizing pancreatitis (ANP). Methods Thirty-three Wistar rats were divided into five groups: normal group, normal saline (NS) group, ANP group, prophylactic and therapeutic groups with NAC randomly. ANP was produced by 3.5% sodium taurocholate retrograde injection. In the prophylactic group, rats received intravenous (i. v.) injection of NAC (300 mg/kg) 1 hour before taurocholate injection and in the therapeutic group, NAC i.v. injection was given 1 hour after sodium taurocholate injection. Animals were sacrificed at 12 hours after induction of pancreatitis. Activation of NF-KB in pancreatic tissues was determined by electrophoretic mobility shift assay(EMSA). Cell apoptosis was assessed by modified TUNEL method. The following parameters were also measured: plasma levels of amylase and lipase, pancreatic wet/dry weight ratio and histologic grading. Results Taurocholate pancreatitis is characteristic of necrosis, haemorrhage, and severe leukocyte infiltration in the pancreas. Plasma amylase and lipase levels, pancreatic wet/dry weight ratio increased in rats of ANP. NF-κB banding activity was higher after pancreatitis induction (6.03±0.41). When NAC was given 1 hour before induction of pancreatitis, the activation (3.28±0.42) of NF-κB was prevented with significantly decreased severity of pancreatitis as assessed by amylase, pancreatic wet/dry weight ratio. The number of apoptotic cells in pancreatic tissue sections was greater in rats treated with NAC than in rats not treated with NAC. There was a negative correlation between NF-κB banding activity and apoptosis of pancreatic cells (r=-0.96, P< 0.01) and there was a positive correlation between NF-κB activation and histopathological score (r=0.63, P<0.01). Histopathological score of pancreatic injury had negative correlation with apoptosis of pancreatic cells(r=-0.98, P<0.01). Conclusion Early blocked NF-κB activation with NAC increases cell apoptosis in pancreatic tissue and decreases edema of pancreas and severity of pancreatitis in rats with ANP. Key words: Pancreatitis, acute necrotizing; Models, animal; NF-κB; Cell apoptosis

Objective To investigate the aggravation of pancreatic tissue injury in rats with acute hypertriglyceridemic pancreatitis and the possible role of NADPH oxidase (NOX). Methods Thirty SPF rats were randomly (random number)divided into five groups: N group, H group, NLAP group, HLAP group and HAPO group. AMY, TG, TC and FFA levels were detected. The pathological changes of pancreas were observed under light microscope and the ultrastructural changes of pancreatic acinar cells were observed by TEM. Serum levels of MDA, SOD, IL-1β, TNF-α and LDH were detected. The expression of NOX4, p-Akt and p-GSK3β in pancreas was detected by immunofluorescence, and the expression of NF-κB and TNF-α in pancreas was detected by immunohistochemistry. Results Intraperitoneal injection of P-407 could significantly increase the levels of serum TG, TC and FFA in rats. After acute pancreatitis induced by L-Arg, the levels of serum AMY in the NLAP and HLAP groups were significantly increased, while Apocynin could significantly decrease the level of serum AMY. Compared with the NLAP group, the pathological injury of pancreatic tissue in the HLAP group was more serious, the level of inflammatory mediators was significantly increased, and the cell necrosis was more serious. After inhibiting NOX, the activation of Akt/GSK3β pathway was regulated and the pancreatic injury was improved. Conclusion In HTGP, NOX aggravates pancreatic injury by regulating the activation of Akt/GSK3 β pathway. Inhibition of NOX expression can play a protective role in pancreas injury of HTGP.. Key words: Hypertriglyceridemia; Acute pancreatitis; Pancreatic injury; NADPH oxidase; Oxidative stress

Serum microRNAs-217 and -375 as biomarkers of acute pancreatic injury in rats.

Background: In trauma patients, pancreatic injury is rare; however, if undiagnosed, it is associated with high morbidity and mortality rates. Few predictive models are available for the identification of pancreatic injury in trauma patients with elevated serum pancreatic enzymes. In this study, we aimed to construct a model for predicting pancreatic injury using a decision tree (DT) algorithm, along with data obtained from a population-based trauma registry in a Level I trauma center. Methods: A total of 991 patients with elevated serum levels of amylase (>137 U/L) or lipase (>51 U/L), including 46 patients with pancreatic injury and 865 without pancreatic injury between January 2009 and December 2016, were allocated in a ratio of 7:3 to training (n = 642) or test (n = 269) sets. Using the data on patient and injury characteristics as well as laboratory data, the DT algorithm with Classification and Regression Tree (CART) analysis was performed based on the Gini impurity index, using the rpart function in the rpart package in R. Results: Among the trauma patients with elevated amylase or lipase levels, three groups of patients were identified as having a high risk of pancreatic injury, using the DT model. These included (1) 69% of the patients with lipase level ≥306 U/L; (2) 79% of the patients with lipase level between 154 U/L and 305 U/L and shock index (SI) ≥ 0.72; and (3) 80% of the patients with lipase level <154 U/L with abdomen injury, glucose level <158 mg/dL, amylase level <90 U/L, and neutrophil percentage ≥76%; they had all sustained pancreatic injury. With all variables in the model, the DT achieved an accuracy of 97.9% (sensitivity of 91.4% and specificity of 98.3%) for the training set. In the test set, the DT achieved an accuracy of 93.3%, sensitivity of 72.7%, and specificity of 94.2%. Conclusions: We established a DT model using lipase, SI, and additional conditions (injury to the abdomen, glucose level <158 mg/dL, amylase level <90 U/L, and neutrophils ≥76%) as important nodes to predict three groups of patients with a high risk of pancreatic injury. The proposed decision-making algorithm may help in identifying pancreatic injury among trauma patients with elevated serum amylase or lipase levels.

Octreotide prevents l-asparaginase–induced pancreatic injury in rats

Diabetes is usually associated with oxidative stress that causes hepatic and pancreatic tissue injury. This work was carried out to evaluate the effect of Cucumis sativus and Cucurbita maxima methanol extracts on the streptozotocin-induced diabetic hepatic and pancreatic injury in rats. Diabetes was induced in seven equal groups of rats by a single intraperitoneal injection of streptozotocin (40 mg/kg), in addition to the non-diabetic control group. Two diabetic groups were treated with Cucumis sativus methanol extract and two were treated with Cucurbita maxima, each at 200 and 400 mg/kg for 21 days after streptozotocin-induced diabetes. Another diabetic group was treated with both Cucumis sativus and Cucurbita maxima at 200 mg/kg of each. Another group was treated with metformin (200 mg/kg orally). The plant extracts normalized serum liver enzymes activities, oxidative stress markers, and restored serum proteins and lipid profile. They also significantly reduced blood sugar to values comparable to non-diabetic rats. The hypoglycemic effect is also confirmed by the improvement of the immunohistochemical expression of insulin in β-cells of islets of Langerhans. Hepatic and pancreatic protection was also confirmed by the improvement of the histopathological picture as compared to STZ-diabetic rats. The GC-MS analysis revealed the presence of 35 and 34 compounds in the methanol extract of cucumber and pumpkin, respectively. Finally, the methanol extract of cucumber and pumpkin could be beneficial acting synergistically in the protection of the liver and pancreas against diabetes-induced tissue damage.

Several chemicals such as N-diethylnitrosamine (DEN) promote hepatocellular cancer in rodents and induce hepatocyte injury. DEN affects the initiation stage of carcinogenesis together with enhanced cell proliferation accompanied by hepatocellular necrosis. DEN-induced hepatocellular necrosis is reported to be related to enhanced generation of reactive oxygen species. Carnosine (CAR), taurine (TAU), and betaine (BET) are known to have powerful antioxidant properties. We aimed to investigate the effects of CAR, TAU, and BET pretreatments on DEN-induced oxidative stress and liver injury in male rats. Rats were given CAR (2 g L-1 in drinking water), TAU (2.5% in chow), and BET (2.5% in chow) for 6 weeks and DEN (200 mg kg-1 intraperitoneally) was given 2 days before the end of this period. Serum alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and γ-glutamyl transferase activities were determined and a histopathologic evaluation was performed on the liver tissue. Oxidative stress was detected in the liver by measuring malondialdehyde, diene conjugate, protein carbonyl and nitrotyrosine levels, glutathione and glutathione peroxidase levels, and superoxide dismutase and glutathione transferase activities. Pretreatments with CAR, TAU, and BET decreased liver prooxidant status without remarkable changes in antioxidant parameters in DEN-treated rats. Pretreatments with TAU and BET, but not CAR, were also found to be effective to reduce liver damage in DEN-treated rats. In conclusion, TAU, BET, and possibly CAR may have an ameliorating effect on DEN-induced hepatic injury by reducing oxidative stress in rats.

Fatty Acid Ethyl Esters–Alcohol’s Henchmen in the Pancreas?

Background/Aims: Dachengqi decoction (DCQD) is a well-known traditional Chinese herbal drug with strong anti-inflammatory effects. Angiopoietin-1 (Ang-1) plays a vital role in maintaining the stability and integrity of the vascular wall and prevents vascular leakage due to inflammatory mediators. Our previous work found that DCQD protects against pancreatic injury in rats with severe acute pancreatitis (SAP). This study aims to investigate the effects of DCQD on intestinal endothelial damage in both damaged human umbilical vein endothelial cells (HUVECs) and SAP rats. Methods: HUVECs were randomly divided into four groups: control group, TNF-α group, TNF-α plus Ang-1 group (Ang-1 group), and TNF-α plus DCQD group (DCQD group). Cells were incubated for 6 h, 12 h, and 24 h, before collection. The treatment concentration of DCQD was decided based on a Cell Counting Kit-8 (CCK-8) assay. The monolayer permeability of the HUVECs was assessed by measuring the transendothelial electrical resistance (TEER). Apoptosis was analyzed by flow cytometry. mRNA and protein expression of aquaporin 1 (AQP-1), matrix metalloproteinase 9 (MMP9), and junctional adhesion molecule-C (JAM-C) was evaluated by RT-PCR, immunocytofluorescence, and western blot. Forty male Sprague-Dawley rats were randomized into a control group, SAP group, SAP plus Ang-1 group (Ang-1 group), and SAP plus DCQD group (DCQD group). SAP was induced by intraperitoneal injection of cerulein and lipopolysaccharide (LPS), while the control group received 0.9% saline solution. Evans blue was injected through the penile vein and the rats were then sacrificed 12 h after modeling. Levels of serum amylase, TNF-α, IL-1β, IL-2, and IL-6 were determined by using ELISA. Intestinal tissue was analysed by histology, and capillary permeability in the tissues was evaluated by Evans blue extravasation assay. Protein and mRNA expression of AQP-1, MMP9, and JAM-C were assessed by immunohistofluorescence, western blot, and RT-PCR. Results: DCQD reduced the permeability of HUVEC induced by TNF-α in vitro. Furthermore, DCQD altered the mRNA and protein levels of JAM-C, MMP9, and AQP-1 in HUVECs after TNF-α induction. SAP intestinal injury induced by cerulein combined with lipopolysaccharides was concomitant with increased expression of JAM-C and MMP9, and reduced expression of AQP-1 in intestinal tissue. Pretreatment with DCQD attenuated SAP intestinal injury and lowered the levels of serum amylase, TNF–α, IL-1β, IL-2, and IL-6 effectively. Our study demonstrated that DCQD decreased the expression of JAM-C and MMP9 and increased the expression of AQP-1 both in vitro and in vivo. Conclusion: DCQD can reduce capillary endothelial damage in acute pancreatitis-associated intestinal injury and the mechanism may be associated with the regulation of endothelial barrier function-associated proteins AQP-1, MMP9, and JAM-C.

Utility of serum pancreatic enzyme levels in diagnosing blunt trauma to the pancreas: A prospective study with systematic review

Various parts of the Moringa oleifera (M. oleifera) tree are widely accepted to have ameliorative effects against metal toxicity. In the present study, M. oleifeira oil (MO) was tested against HgCl2-induced tissue pathologies and oxidative stress. Male Wistar rats were administered MO (1.798 mg/kg p.o.) or HgCl2 (5 mg/kg body wt) alone or in combination (5 mg/kg HgCl2+1.798 mg/kg MO p.o.) three times per week for 21 days. After exposure and treatment periods, rats were sacrificed; blood collected and the oxidative status of the liver and kidney homogenates were evaluated. In the liver, malondialdehyde (MDA) level, glutathione (GSH), and superoxide dismutase (SOD) activities were higher whereas catalase (CAT) activity was lower in the HgCl2 group than in the control group. In the kidney, MDA level, SOD, and CAT activities were higher whereas GSH activity was unchanged in the HgCl2 group compared to the control group. In the liver, MDA level, SOD, and CAT activities were lower in the HgCl2+MO group than in the HgCl2 group. In the kidney, MDA level, SOD and CAT activities were lower in the HgCl2+MO than in the HgCl2 group. Furthermore, Hg-induced increases in creatinine and bilirubin levels as well as the increase in γ-glutamyl transferase, lactate dehydrogenase, and alkaline phosphatase activities were attenuated in the combine exposure group and the animals showed improvement in the histology of the liver and kidney. MO decreased the negative effects of Hg-induced oxidative stress in rats.

Background: Cadmium can induce pancreatic injury via oxidative stress, inflammation and apoptosis. Naringenin (NGN) and vanillin (VLN) exert antioxidant, anti-inflammatory, and antiapoptotic effects. Objective: The likely ameliorative effects of NGN, VLN and their combination were studied in rats exposed to cadmium-induced pancreatic injury. Materials and Methods: Rats received NGN (50 mg/kg/day, p.o.), VLN (100 mg/ kg/day, p.o.), or NGN + VLN for 7 days and one injection of CdCl2 (2 mg/kg, i.p.) on the 6th day. Results: Cadmium significantly lowered serum amylase and insulin levels. Cadmium also caused significant increments of malondialdehyde, tumor necrosis factor-α, interleukin-1β, nuclear factor-κB p65, Bax/Bcl-2 ratio and phosphorylated c-Jun N-terminal kinase (p-JNK) and p38 mitogen-activated protein kinases (MAPKs) and significant decrements of reduced glutathione and catalase in the pancreas of rats received CdCl2. Additionally, CdCl2 caused marked histopathological necrosis and significantly increased caspase-3 expression in pancreatic tissue. The cadmium-induced biochemical, histopathological and immunohistochemical changes were significantly ameliorated by NGN, VLN and NGN + VLN. However, NGN + VLN caused more significant ameliorative effects than did NGN and VLN alone. Conclusion: NGN, VLN and NGN + VLN afforded significant protection of pancreas in rats exposed to cadmium insult through modulation of JNK and p38 MAPK pathways and inhibition of oxidative stress, inflammation and apoptosis.

An increase in systemic inflammatory mediators from stimulated leucocytes and macrophages has been noted during acute pancreatitis. The role of cytolytic inflammatory macrophages and potential mechanisms in the development of acute pancreatic injury and endothelial barrier dysfunction are less well defined. Rats were challenged by an intraperitoneal injection of cytolytic or non-cytolytic inflammatory macrophage stimulators at various concentrations. The effects of oxygen free radicals, prostaglandin and extracellular calcium influx on macrophage-associated pancreatic endothelial compromise, measured by pancreatic intravascular plasma volume, pancreatic interstitial fluid volume, and the pancreatic extravascular human serum albumin distribution volume, were explored. Zymosan-induced overactivation of cytolytic inflammatory macrophages resulted in the development of acute pancreatic endothelial dysfunction in a dose- and time-dependent pattern. An increase in pancreatic water content and interstitial fluid volume was observed following a higher dose (0.5 mg/g) of concanavalin A without alteration in plasma lipase level, while thioglycollate medium did not compromise pancreatic endothelial barrier function. Oxygen free radicals, but also prostaglandins and extracellular calcium influx, seemed to be involved in macrophage overactivation-induced pancreatic injury. Overactivation of cytolytic macrophages plays a role in the pathogenesis of pancreatic injury by initiating the development of endothelial barrier dysfunction. Multiple inflammatory mediators from overactivated macrophages act as intercellular signals between macrophages and the endothelium during acute pancreatic injury.

This study was designed to evaluate the protective effect of the cysteinyl leukotriene receptor antagonist montelukast against pancreatic injury during acute pancreatitis. Acute pancreatitis was induced in rats by 20-μg/kg (intraperitoneal) cerulein given at 1-hour intervals within 4 hours. Montelukast was administered intraperitoneally at a dose of 10 mg/kg 15 minutes before the first cerulein injection. Six hours after the cerulein or saline injections, the animals were killed by decapitation. Blood samples were collected to analyze amylase, lipase, and the proinflammatory cytokines tumor necrosis factor α and interleukin 1β. Pancreas tissues were taken for the determination of tissue glutathione and malondialdehyde levels and Na,K-adenosine triphosphatase and myeloperoxidase activities. The extent of tissue injury was analyzed microscopically. Acute pancreatitis caused significant decreases in tissue glutathione level and Na,K-adenosine triphosphatase activity, which were accompanied with significant increases in the pancreatic malondialdehyde level, myeloperoxidase activity, and plasma cytokine level. On the other hand, montelukast treatment reversed all these biochemical indices and histopathological alterations that were induced by cerulein. These results suggest that cysteinyl leukotrienes may be involved in the pathogenesis of acute pancreatitis and that the cysteinyl leukotriene receptor antagonist, montelukast, might be of therapeutic value for treatment of acute pancreatitis.

Effects of gender differences and endurance training on exhaustive exercise induced-oxidative stress have been a question that has not been clarified in the literature. The aim of this study was to determine the effects of sex, acute exhaustive exercise and chronic aerobic exercise training on oxidative stress in the heart and the skeletal muscle. The study was carried out with 12 week-old male (n = 24) and female (n = 24) young adult Wistar rats. They were randomly divided into four groups: untrained, trained, untrained exhausted and trained exhausted. The rats in the trained group swam for 60 min/day, five days per week for eight weeks. Thereafter, one-half of the trained and one-half of the untrained rats were randomly selected into the trained and untrained exhaustive exercise groups, respectively. They were killed immediately after one last exhaustive swimming exercise. In the heart, endurance training decreased malondialdehyde (MDA) levels in the female rats at rest, but did not change in the male rats in the heart; MDA levels were also increased in female rats at rest in the gastrocnemius tissues. [corrected]. In the trained female rats, exhaustive exercise decreased MDA levels in the heart and gastrocnemius tissues. The nitric oxide (NO) levels in the heart in the untrained female rats were higher than in the male rats after exhaustive exercise. Training decreased the NO levels in both sexes in the gastrocnemius tissue at rest. In the heart, the untrained female rats had higher total glutathione (GSH) levels than in the male rats at rest. Also, exhaustive exercise decreased the GSH levels in the trained female rats. In the gastrocnemius, untrained female rats showed higher GSH levels than in the male after exhaustive exercise. The superoxide dismutase activities in the gastrocnemius were similar between the female and male rats. The results suggested that gender was a major determinant of changes in MDA, NO and GSH levels in the heart and gastrocnemius tissues after the exhaustive exercise or endurance training. Also, the responses to oxidative stress induced by acute exercise or training in the heart and gastrocnemius muscle tissues are different.