bromochloroacetic-acid and Reperfusion-Injury

The ischemia-reperfusion injury of the small intestine is a condition of high mortality, which occurs following superior mesenteric artery (SMA) embolization or circulatory redistribution. The aim of the study was to evaluate the local and systemic effects of postconditioning in a rat model of small intestine ischemia-reperfusion.. Male Wistar rats underwent 60 min ischemia by the clamping of the SMA, followed by 6 hrs of reperfusion. The animals (n = 30) were randomized into three groups: sham-operated, control-, and postconditioned. Postconditioning was performed at the very onset of reperfusion by 6 alternating cycles of 10-10 seconds reperfusion/reocclusion, for a total of 2 min. At the end of the reperfusion blood and tissue (small intestine, lungs, kidney, liver) samples were taken for histological examination. The antioxidant status of small intestine was measured from intestinal homogenates.. Histologic results revealed increased damage in control-group lungs, kidney, liver and small intestine in comparison with the postconditioned group. The injury was supported by significantly higher wet/dry weight ratio (p = 0.026), and serum levels of creatinine (p = 0.013), ASAT (p = 0.038), LDH (p = 0.028) and CK (p = 0.038) in the control group. The postconditioned group showed lower serum IL-6 levels (420 pg/ml vs. 188 pg/ml), as well as significantly higher mucosal antioxidant concentration.. Postconditioning was able to decrease not only local, but the systemic damage intensity also, after a small intestinal ischemic-reperfusion episode.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Biomarkers; Cholesterol, HDL; Creatinine; Enzyme-Linked Immunosorbent Assay; Intestine, Small; Ischemic Postconditioning; Keratins; Kidney; Liver; Lung; Male; Mesenteric Artery, Superior; Organ Size; Random Allocation; Rats; Rats, Wistar; Reperfusion Injury

Ischemia-reperfusion (I/R) injury in liver transplantation units and the influence of I/R injury on bile flow dynamics is being intensely investigated in various animal models, but the expression of intracellular intermediate filaments of biliary type as an early sign of cholestasis has not been yet explored.. We studied the hepatic elimination kinetics of indocyanine green (ICG), an exclusively biliary excreted cholephilic dye, and the functional and morphological integrity of liver cells in a canine liver transplantation model following I/R. During reperfusion following cold ischemia, we evaluated the ICG excretion curves, biochemical signs of liver damage, the bile canaliculus of the hepatocytes by electron microscopy, and the expression of intermediate filaments of cytokeratin type by immunohistochemistry.. Impairment of the biliary ICG excretion was directly related to ischemia time, but hepatocellular ICG uptake and bile flow rate were not significantly reduced. Liver enzymes increased as early as 6 h of ischemia and hepatocytes showed an increase of the bile canaliculus area. This was correlated to a membranous to cytoplasmatic staining of the cytoskeleton of the hepatocytes.. To the best of our knowledge, this is the first evidence of cholestatic changes starting early following cold ischemia in a canine isolated perfused liver transplantation model despite prompt recovery of the bile flow.

Topics: Animals; Bile; Cholestasis; Coloring Agents; Disease Models, Animal; Dogs; Hepatocytes; Immunohistochemistry; Keratins; Liver; Liver Transplantation; Microscopy, Electron; Reperfusion Injury

Kidney is thought to be a regenerative organ in terms of repair from acute tubular injury. It is unknown whether cell population contributes to repair disordered kidney. We attempted to identify and isolate highly proliferative cells from a single cell. We dissected a single nephron from adult rat kidney. Isolated nephrons were separated into segments and cultured. Outgrowing cells were replated after limiting dilution so that each well contained a single cell. One of cell line which was the most potent to grow was designated as rKS56. rKS56 cells showed cobblestone appearance and expressed immature cell markers relating to kidney development and mature tubular cell markers. rKS56 cells grew exponentially and could be maintained for 300 days without transformation. In different culture conditions, rKS56 cells differentiated into mature tubular cells defined by aquaporin-1, 2 expression, and responsiveness to parathyroid hormone or vasopressin. Engrafted to kidney in rat ischemic reperfusion model, rKS56 cells replaced in injured tubules in part after implantation and improved renal function. These results suggest rKS56 cells possess character such as self-renewal, multi-plasticity and capability of tissue repair. rKS56 may possibly contribute to the future development of cell therapy for renal regeneration.

Topics: Animals; Aquaporin 1; Aquaporin 2; Arginine Vasopressin; Cell Culture Techniques; Cell Differentiation; Cell Line; Cell Proliferation; Cells, Cultured; Cyclic AMP; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Immunoblotting; Karyotyping; Keratins; Kidney; Kidney Diseases; Kidney Tubules; Male; Microscopy, Fluorescence; Nephrons; Parathyroid Hormone; Phenotype; Rats; Rats, Sprague-Dawley; Regeneration; Renal Insufficiency; Reperfusion Injury; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stem Cells; Time Factors; Vasopressins

In orthotopic liver transplantation, ischemic-reperfusion is one of the most important factors that cause the incidence of biliary compliance. The aim of this study was to investigate the effects of ischemia reperfusion on epithelial cells apoptosis and proliferation of intrahepatic bile duct (IBD) (>20 microm).. 30-minute warm ischemia was applied to rat livers respectively, and experiment was performed on days 2, 7, 14, 28 after reperfusion. Apoptosis was determined in situ by morphology and TUNEL, and cholangiocyte proliferation was evaluated in situ by morphometry of liver sections stained for cytokeratin-19 (CK-19) and by proliferating cellular nuclear antigen staining in liver sections.. Two days after ischemia reperfusion, apoptosis of cells was observed in large intrahepatic bile ducts (>20 microm) (5.6%+/-1.2%), but the number of large intrahepatic bile ducts reduced (0.32+/-0.06). Seven days after ischemia reperfusion, the apoptosis index of cholangiocytes decreased to 1.2%+/-0.3%, and the number of intrahepatic bile ducts began to proliferate and returned to nearly normal on day 28.. Ischemia reperfusion causes a decrease in the number of intrahepatic bile ducts (>20 microm) as a result of a higher rate of apoptosis and absence of initial proliferation.

Topics: Animals; Apoptosis; Bile Ducts, Intrahepatic; Cell Division; Epithelial Cells; Hepatocytes; Immunohistochemistry; In Situ Nick-End Labeling; Keratins; Male; Necrosis; Proliferating Cell Nuclear Antigen; Rats; Rats, Sprague-Dawley; Reperfusion Injury

Apoptosis is a cellular suicide program that can be activated by cell injury or stress. Although a number of laboratory studies have shown that ischemia/reperfusion injury can induce apoptosis, few clinical studies have been performed. The purpose of this study was to determine whether apoptosis is a major mechanism of cell death in intestinal epithelial cells and lymphocytes in patients who sustained trauma, shock, and ischemia/ reperfusion injury.. Intestinal tissues were obtained intraoperatively from 10 patients with acute traumatic injuries as a result of motor vehicle collisions or gun shot wounds. A control population consisted of six patients who underwent elective bowel resections. Apoptosis was evaluated by conventional light microscopy, laser scanning confocal microscopy using the nuclear staining dye Hoechst 33342, immunohistochemical staining for active caspase-3, and immunohistochemical staining for cytokeratin 18.. Academic medical center.. Patients with trauma or elective bowel resections.. Extensive focal crypt epithelial and lymphocyte apoptosis were demonstrated by multiple methods of examination in the majority of trauma patients. Trauma patients having the highest injury severity score tended to have the most severe apoptosis. Repeat intestinal samples obtained from two of the trauma patients who had a high degree of apoptosis on initial evaluation were negative for apoptosis at the time of the second operation. Tissue lymphocyte apoptosis was associated with a markedly decreased circulating lymphocyte count in 9 of 10 trauma patients.. Focal apoptosis of intestinal epithelial and lymphoid tissues occurs extremely rapidly after injury. Apoptotic loss of intestinal epithelial cells may compromise bowel wall integrity and be a mechanism for bacterial or endotoxin translocation into the systemic circulation. Apoptosis of lymphocytes may impair immunologic defenses and predispose to infection.

Topics: Adolescent; Adult; Apoptosis; Caspase 3; Caspases; Cell Death; Epithelial Cells; Female; Humans; Intestinal Mucosa; Ischemia; Keratins; Lymphocytes; Male; Middle Aged; Multiple Trauma; Reperfusion Injury; Shock