Hepatic Cell Integrity Research Articles

Effect of garden cress in reducing blood glucose, improving blood lipids, and reducing oxidative stress in a mouse model of diabetes induced by a high-fat diet and streptozotocin.

A mouse model in which diabetes mellitus was induced by low-dose streptozotocin (STZ) injection combined with a high-fat diet was used to study the effect of two water cress (Lepidium savitum) preparations. Diabetic mice were treated with dried cress powder or with water-soluble extracts (tested at two doses), together with proper control groups. The mice were evaluated after 4 weeks of continuous intervention for type 2 diabetic and associated markers. We determined blood glucose, body weight, total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), serum insulin levels, and DNA integrity of hepatic cells. The concentrations of malondialdehyde (MDA) and lipid peroxide (LPO) and the activities of four enzymes that are part of the antioxidant defense system were determined in liver samples, as well as gene expression (by semi-quantitative reverse transcription polymerase chain reaction) and enzyme activity of IRS-1, IRS-2, PI3K, AKT-2, and GLUT4. After 4 weeks of intervention, the levels of TC, TG, and LDL cholesterol were significantly (P < 0.5) decreased and HDL cholesterol was significantly increased. Enzyme activities of liver superoxide dismutase, glutathione, glutathione peroxidase, and catalase were significantly increased, whereas MDA and LPO concentrations were significantly reduced. The transcription level of the five genes assessed was increased, with corresponding increases in protein expression. Oral uptake of garden cress can significantly reduce the blood glucose and improve the blood lipid metabolism of diabetic mice. Considerable improvements in the activity of antioxidant defense enzymes were observed in type 2 diabetic mice that improved the body's antioxidant emergency response. © 2019 Society of Chemical Industry.

Osteopathic lymphatic techniques reduces cortisol plasma level in rats after repeated treatments

BackgroundMany reports evidence in humans the positive effect of manipulation, in particular of osteopathic lymphatic techniques in normal as well in pathological conditions. In animals too, lymphatic treatment has been reported to induce changes of lymphatic flux and to mobilize inflammatory mediators thus determining positive effects on the immune system. MethodsTwelve male Wistar rats (weight 180 g) were randomly allocated to control (6 rats) and experimental (6 rats) group. Experimental group rats underwent lymphatic techniques for 10 min each day for 5 consecutive days under gaseous anaesthesia. Control group animals were anaesthetized but did not have lymphatic treatment. Blood samples were collected before and at the end of the experiment period. A complete blood cell count and serum chemistry were determined. TNF-alpha and IL-1 beta were determined in order to evaluate changes in inflammatory response. Cortisol was also determined as indicative of animal stress. The experiment was carried on in August 2011 and was replicated in February 2012. ResultsBlood cell count did not show differences at baseline and at the end of the experiment both in control and in treated rats. Blood serum chemistry was also unchanged. In particular, blood urea nitrogen (BUN) and serum creatinine (expression of renal function), alanine transaminase (ALT) and aspartate transaminase (AST) (indicative of hepatic cell integrity) were normal and did not change after lymphatic treatment. No changes in plasmatic level of cytokines were found after the five-days lymphatic treatment. On the contrary and unexpectedly, cortisol level was significantly reduced in rats that were treated with lymphatic techniques. In untreated rat group cortisol level was, however, minimally increased. These findings were confirmed by the results of the replicated study. ConclusionsThis study demonstrates that repeated lymphatic treatment is a safe procedure. Moreover, lymphatic techniques induce reduction in cortisol level, which may be related to an anti-stress effect.

Butea monosperma and chemomodulation: Protective role against thioacetamide-mediated hepatic alterations in Wistar rats

The present study was carried out to study the effect of Butea monosperma, a known liver acting drug on the tumor promotion related events of carcinogenesis in rat liver. Thioacetamide (TAA) was used to induce tumor promotion response and oxidative stress and caused significant depletion in the detoxification and antioxidant enzyme armory with concomitant elevation in malondialdehyde (MDA) formation, hydrogen peroxide (H(2)O(2)) generation, ornithine decarboxylase (ODC) activity and unscheduled DNA synthesis. However, B. monosperma pretreatment at two different doses restored the levels of the above-said parameters (p < 0.001) in a dose-dependent manner. The alcoholic extract of B. monosperma used in the present study seems to offer dose-dependent protection and maintain the structural integrity of hepatic cells. This was evident from the significant reduction in TAA-induced serum GOT, GPT, Lactate dehydrogenase (LDH) and gamma-Glutamyl transpeptidase activity (GGT) activities (p < 0.001). These investigations validate the use of B. monosperma in liver disorders by Ayurvedic physicians. Overall results indicate that the methanolic extract of B. monosperma possesses hepatoprotective effects and also it might suppress the promotion stage via inhibition of oxidative stress and polyamine biosynthetic pathway.

Hepatic Function Is Preserved Following Liver-Directed, Adenovirus-Mediated Gene Transfer

Inherited and acquired disorders of the liver are attractive targets for gene therapy. Hepatic cells are susceptible targets for shuttle vectors because of a diversity of protein and viral receptors and accessibility of a selective afferent blood supply. Preservation of existing hepatic cell integrity and metabolic function is of paramount importance for successful whole animal gene therapy trials. In this report, we examine hepatic cell function and integrity following adenovirus-mediated reporter gene transfer to the liver in vivo. E1-deleted, replication-defective adenovectors encoding the LacZ gene driven by the human CMV promoter were delivered to the liver by isolated portal perfusion. The gene transfer rate, as determined by specific histochemical staining, approached 30% with recombinant protein detectable by Western blot throughout the course of study. Hepatic cell integrity as assessed by histology and hepatic enzyme profile (serum aspartate aminotransferase, γ-glutamyl transpeptidase) demonstrated normal cellular architecture and no significant difference between transfected liver and controls. Hepatic synthetic and metabolic function, as determined by albumin levels, prothrombin time, and bilirubin, were similar between the two study groups. This study demonstrates that efficient adenovirus-mediated gene transfer and expression in the rat liver do not compromise hepatic cell metabolism and integrity.

In vivo studies on toxic effects of concurrent administration of paracetamol and its [formula omitted] ester (SUR 2647 combination)

1. 1. Single p.o. doses of paracetamol 400 and 800 mg/kg or SUR 2647 combination (free paracetamol + paracetamol-N- acetyl- dl-methionate , paracetamol/methionine ratio 2:1) equivalent to paracetamol 400 and 800 mg/kg were given to Bom:NMRI mice. Vehicle treated (1% w/v aqueous methylcellulose) mice were established as a control group. 2. 2. All treatment groups irrespective of medication caused an initial GSH depletion. However, SUR 2647 combination 400 mg/kg caused a much earlier hepatic GSH recovery than paracetamol 400 mg/kg. SUR 2647 combination 800 mg/kg caused a higher hepatic GSH level than paracetamol 800 mg/kg. 3. 3. There was no significant difference in the plasma ALAT level after SUR 2647 combination 400 or 800 mg/kg and the control group. Paracetamol 400 and 800 mg/kg caused significant plasma ALAT elevations compared to the control group. 4. 4. The addition of N- acetyl- dl-methionine esterified to paracetamol, as in the SUR 2647 combination, enhances the hepatic GSH synthesizing capacity in Bom:NMRI mice after experimental overdosage and offers protection of hepatic cell integrity as assessed by plasma ALAT level compared to paracetamol alone.

Efficacy of paracetamol-esterified methionine versus cysteine or methionine on paracetamol-induced hepatic GSH depletion and plasma alat level in mice

The effect of paracetamol- N- acetyl- dl-methionate (PAM) in preventing paracetamol-induced hepatic glutathione (GSH) depletion and hepatic cell damage assessed by plasma ALAT level, was compared to those of concomitantly administered paracetamol and N- acetyl- l-cysteine (NAC) or N- acetyl- dl-methionine (NAM) and paracetamol 400 mg/kg (P) alone. PAM, NAM and NAC reduced hepatic GSH depletion compared to P. The concomitant administration of GSH precursors in either form apparently maintained hepatic cell integrity as evaluated by plasma ALAT compared to predose and 16 hr control measurements. No statistically significant difference between PAM, NAM and NAC was observed. In group P a statistically significant, but transitory, rise in plasma ALAT level following dosage was seen. NAC was more effective than PAM and NAM in the prevention of GSH depletion 1 hr after dosing but was less effective in promoting de novo GSH synthesis towards 16 hr. There was no statistically significant difference between PAM and NAM with respect to effect on GSH depletion or hepatic cell integrity. PAM and NAM increased the GSH level significantly above control level 16 hr after dosing. PAM is rapidly cleaved to paracetamol and methionine following dosage as shown by the observed plasma paracetamol level. PAM compares favourably in hepatoprophylactic effect, to concomitant administration of equimolar doses of free N- acetyl- dl-methionine added to the paracetamol formulation.

Preservation of hepatic cell integrity by aprotinin in hypoxia

Perfused cat livers subjected to 2.5 hr of hypoxia exhibited dramatic increases in perfusate cathepsin D and lactic acid dehydrogenase (LDH) activities, amino nitrogen concentrations, and a 60% depression in the clearance rate of carbon particles by the reticuloendothelial system (RES). Addition of aprotinin (250 KIU/ml) to the perfusate prior to hypoxia prevented the increases in circulating cathepsin D, LDH, and amino-nitrogen observed at 150 minutes. In addition, aprotinin prevented the reduction in carbon clearance during severe hypoxia. However, aprotinin had no effect on the percent free cathepsin D activity indicating that this agent did not directly prevent increases in lysosomal fragility occurring in response to hypoxia. Thus, addition of pharmacologic doses of aprotinin to the perfusate protected RES cells, and markedly reduced the release of cytoplasmic and lysosomal enzymes. The prevention of cell membrane dissolution appears to be a critical factor in hepatic preservation, and may be related to the inhibition of proteolysis by aprotinin. These effects may help explain the therapeutic effectiveness of this agent in shock and in myocardial ischemia.

Hepatic cell integrity in hypodynamic states.

Changes in liver integrity were studied in isolated perfused cat livers during simulated shock conditions (i.e., combined hypoxia, acidosis, and ischemia) or under the influence of each hypodynamic state separately. The combined hypodynamic stimuli depressed carbon clearance 51% and significantly elevated lactic acid dehydrogenase (LDH) and cathepsin D activities in the perfusate. The perfused liver was more seriously affected by hypoxia than by acidosis or ischemia alone. Reticuloendothelial clearance was depressed 20% and 25% in acidosis and hypoxia, respectively. Hypoxia also induced a 3-fold increase in cathepsin D and a 13-fold increase in LDH activities in the perfusate. After 150 min of hypoxia or ischemia, free cathepsin D in liver tissue increased significantly. The impairment of liver cell integrity (i.e., of Kupffer and parenchymal cells) occurred between 60 and 90 min during simulated shock conditions, indicating that the liver is stable for 60 min when it is exposed to hypoperfusion. The perfused liver is sensitive to local stimuli that predominate in circulatory shock, particularly hypoxia. These stimuli promote the release of lysosomal and cytoplasmic enzymes as well as depress phagocytosis by the reticuloendothelial system, phenomena that exacerbate the shock state.

Pancreatic hydrolases and the formation of a myocardial depressant factor in shock.

Pancreatic hydrolases and the formation of a myocardial depressant factor in shock.

Use of Histochemical Stains in Needle Biopsy of the Liver

Summary Needle biopsy specimens of the liver fixed in absolute alcohol readily lend themselves to staining with both routine hematoxylin and eosin stain and with the periodic acid stain. The latter stain demonstrates quantitative and distributional changes of neutral polysaccharide (glycogen) within the liver cells. Through the application of the neutral polysaccharide stain, it is possible to gain added information regarding hepatic cell integrity. Normally, hepatocellular glycogen is present in generous amounts and is evently distributed throughout the parenchymal cells. In various disease states, where study of hepatic morphology may be unrevealing, reduction in quantity and irregular distribution of intracellular glycogen demonstrated by this stain may be considered evidence of continued disease activity. Liver needle biopsy specimens have routinely been interpreted on the basis of morphologic change. The addition of the neutral polysaccharide stain permits interpretation on the basis of a functional alteration as well, and gives an added opportunity for the correlation of hepatocellular structure with function.