benzyloxycarbonylvalyl-alanyl-aspartyl-fluoromethyl-ketone and Body-Weight

Enterocytes die during high-dose radiation exposure in radiation accidents. The modality of cell death has a profound effect on the therapeutic response. The ilea from mice with 15 Gy total body irradiation (TBI) were drawn, morphological features observed by hematoxylin and eosin staining and transmission electron micrographs. The biochemical features of mouse ileum presented with the structure were cleaved Caspase-3 (apoptosis marker), Light Chain 3 (LC3)-I's conversion to LC3-II (autophagy marker) and high mobility group box chromosomal protein 1's secretion (necrosis marker). Then, the autophagy inhibitor (3-methyladenine), caspase inhibitor (Z-VAD-FMK) or necrosis inhibitor (necrostatin) was used to prevent death. Apoptosis, autophagy and necrosis were all appeared in the ileum, but necrosis had the biggest size; the use of 3-methyladenine and Z-VAD-FMK prolong one day's life of the mice after 15 Gy TBI, necrostatin significantly extended the lifespan of 15 Gy irradiated mice (p < 0.05). The results suggest that the death of enterocytes could not be classified into one type of cell death but rather as 'mixed death.'

Topics: Adenine; Amino Acid Chloromethyl Ketones; Animals; Apoptosis; Autophagy; Body Weight; Caspase 3; Caspase Inhibitors; Enterocytes; Enzyme Inhibitors; Feces; HMGB1 Protein; Imidazoles; Indoles; Intestinal Mucosa; Intestines; Male; Mice, Inbred C57BL; Necrosis; Radiation Dosage; Reactive Oxygen Species; Whole-Body Irradiation

In the initial days after transplantation islets are particularly vulnerable and show increased apoptosis and necrosis. We have studied the effects of caspase inhibition on this early beta cell death in syngeneically transplanted islets. Streptozotocin-diabetic C57BL/6 mice were transplanted with 150 syngeneic islets, an insufficient mass to restore normoglycemia, preincubated with or without the pan-caspase inhibitor z-VAD. fmk 2 h before transplantation. Beta cell apoptosis was increased in control islets on day 3 after transplantation (0.28 +/- 0.02%) compared with freshly isolated islets (0.08 +/- 0.02%, p < 0.001), and was partially reduced in transplanted islets preincubated with z-VAD.fmk 200 microM (0.14 +/- 0.02%, p = 0.003) or with z-VAD.fmk 500 microM (0.17 +/- 0.01%, p = 0.012), but not with a lower z-VAD.fmk (100 microM) concentration. Diabetic mice transplanted with islets preincubated with z-VAD.fmk 500 microM showed an improved metabolic evolution compared with control and z-VAD.fmk 200 microM groups. The z-VAD.fmk 500 microM group showed an overall lower blood glucose after transplantation (p = 0.02), and at the end of the study blood glucose values were reduced compared with transplantation day (15.7 +/- 3.6 vs. 32.5 +/- 0.5 mmol/L, p = 0.001). In contrast, blood glucose was not significantly changed in control and z-VAD.fmk 200 microM groups. Four weeks after transplantation beta cell mass was higher in z-VAD.fmk 500 microM group (0.15 +/- 0.02 mg) than in the control group (0.10 +/- 0.02 mg) (p = 0.043). In summary, the treatment of freshly isolated islets with the caspase inhibitor z-VAD.fmk reduced the subsequent apoptosis of the islets once they were transplanted and improved the outcome of the graft.

Topics: Amino Acid Chloromethyl Ketones; Animals; Apoptosis; Blood Glucose; Body Weight; Caspase Inhibitors; Cysteine Proteinase Inhibitors; Diabetes Mellitus, Experimental; Graft Survival; Islets of Langerhans; Islets of Langerhans Transplantation; Male; Mice; Mice, Inbred C57BL; Time Factors; Tissue Culture Techniques

Malignant mesothelioma (MM) is a cancer with uniformly poor responses to current therapeutic regimens. This study evaluates whether taurolidine, a novel antineoplastic agent, is effective against human MM cell lines and a murine model of human MM.. Cell growth inhibition and viability assays were performed on REN, LRK, and H28 cell lines after 24-72-h exposure to 0-200 microm taurolidine. Cell cycle analysis with annexin-V binding, terminal deoxynucleotidyl transferase-mediated nick end labeling assay, electron microscopy, and response to the general caspase inhibitor z-VAD-fmk were performed on MM cell lines after 24-72-h exposure to 50-150 microm taurolidine. Athymic mice were given i.p. injections of 20 x 10(6) REN cells, followed by i.p. taurolidine (17.5 or 20 mg), 3 days/week for up to 3 weeks. Tumors were assessed at day 30. All statistical tests were two-sided.. A 72-h exposure of MM cells to taurolidine showed IC50 of 28-42.7 microm and 50% viability at 49.8-135 microm. Annexin V assay for apoptosis revealed significant increases in annexin binding after 24-72-h exposure to 50-150 microm taurolidine (P <0.05), which was significantly inhibited by z-VAD (P <0.05). MM cells exposed to 50-150 microm taurolidine for 24-72 h showed terminal deoxynucleotidyl transferase-mediated nick end labeling staining consistent with apoptosis, as well as structural evidence of apoptosis via electron microscopy. In vivo, there were significant tumor reductions (62 to >99% reduction) for all dosage regimens compared with untreated controls (P <0.001). In addition, all control animals exhibited ascites and diaphragmatic tumors while treated animals did not.. Taurolidine has significant antineoplastic activity against MM in vitro and in vivo, in part, due to tumor cell apoptosis. These findings warrant further study for potential clinical usefulness.

Topics: Amino Acid Chloromethyl Ketones; Animals; Annexin A5; Antineoplastic Agents; Apoptosis; Body Weight; Caspases; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; Enzyme Inhibitors; Humans; In Situ Nick-End Labeling; Lung Neoplasms; Male; Mesothelioma; Mice; Mice, Nude; Microscopy, Electron; Phosphatidylserines; Taurine; Thiadiazines; Time Factors

Blockade of angiotensin type 1 (AT1) receptors induces smooth muscle cell (SMC) death and regression of aortic hypertrophy in spontaneously hypertensive rats (SHR). We postulated that SMC death and vascular remodeling in this model may be attenuated by z-Val-Ala-Asp(OMe)-CH2F (z-VAD-fmk), a tripeptide inhibitor of caspase enzymes mediating apoptosis. To determine the time course of SMC death and aortic remodeling, SHR were treated with losartan (30 mg/kg per day) for up to 9.5 days. Transient SMC apoptosis occurred in the aortic media with a peak around day 5 of treatment, with increases in the Bax to Bcl-2 protein ratio (>3-fold), in active caspase-3 (5.6-fold), in TUNEL-positive nuclei (19-fold), preceding by 24 hours the peak activation of capase-9 (3.8-fold), and significant reductions in SMC number (46%) and aortic cross-sectional area (8.5%) at 5.5 days. The decrease in total aortic DNA reached significance at 6.5 days (29%). Blood pressure reduction with losartan was progressive and reached significance at day 7 of treatment. Next, we examined the causal link between vascular apoptosis and remodeling. SHR received placebo or losartan (30 mg/kg per day) for 6 days. During the last 24 hours, a subgroup of losartan-treated rats received 3 IV injections of z-VAD-fmk (cumulative dose: 4.4 mg x kg(-1)). All other rats received the vehicle, DMSO. The 24-hour cotreatment with z-VAD-fmk effectively prevented losartan-induced caspase-3 activation and internucleosomal DNA fragmentation, as well as SMC depletion and the reductions in aortic mass and DNA content. Together, these data suggest that caspase-dependent SMC death mediates the early phase of vascular remodeling in response to AT1 receptor blockade in this model of hypertension.

Topics: Amino Acid Chloromethyl Ketones; Animals; Antihypertensive Agents; Aorta; Apoptosis; bcl-2-Associated X Protein; Blood Pressure; Body Weight; Caspase 3; Caspase 9; Caspase Inhibitors; Caspases; Cell Count; DNA Fragmentation; Hypertension; Hypertrophy; In Situ Nick-End Labeling; Losartan; Male; Muscle, Smooth, Vascular; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Inbred SHR; Remission Induction; Time Factors

Acute fatty degeneration in the liver is caused by various agents, such as aspirin, valproic acid, and ibuprofen, that directly inhibit mitochondrial beta-oxidation of fatty acid and oxidative phosphorylation. Endogenous molecules, such as cytokines and hormones, are also known to mediate microvesicular steatosis in liver failure. In this study, we examined how interleukin-12 (IL-12) and IL-18 cause steatosis in the liver. Administration of these cytokines in combination caused marked hepatosteatosis and weight loss in mice. There were marked increases in levels of interferon-gamma (IFN-gamma), nitrite (NO(2)/NO(3)), and fibrinogen in the circulation in these mice. On the other hand, the ATP concentration and blood flow in the liver were significantly reduced. These changes, except the production of IFN-gamma and NO, were partially inhibited by Z-VAD-fmk, a synthetic tripeptide inhibitor for NO-induced caspases. These results indicate that IL-12 and IL-18 may mediate inflammatory hepatosteatosis through impairment of the microcirculation, which leads to mitochondrial dysfunction in hepatocytes.

Topics: Adenosine Triphosphate; Amino Acid Chloromethyl Ketones; Animals; Body Weight; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Fatty Liver; Female; Fibrinogen; Interferon-gamma; Interleukin-12; Interleukin-18; Laser-Doppler Flowmetry; Liver; Liver Function Tests; Mice; Mice, Inbred C57BL; Mice, Inbred MRL lpr; Mice, Knockout; Mitochondria, Liver; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrites; Oxidation-Reduction; Regional Blood Flow; Time Factors

Inflammatory processes may play an important role in the degeneration of basal forebrain cholinergic cells Alzheimer's disease. We infused the proinflammagen lipopolysaccharide into the basal forebrain of young rats and determined whether the chronic administration of two novel non-steroidal anti-inflammatory drugs or a pan-caspase synthesis inhibitor, z-Val-Ala-Asp(OMe)-fluoromethyl ketone (zVAD), could provide neuroprotection from the cytotoxic effects of the neuroinflammation. Chronic lipopolysaccharide infusions decreased choline acetyltransferase activity and increased the number of activated microglia within the basal forebrain region. The level of caspases 3, 8 and 9 was increased in ventral caudate/putamen. Non-steroidal anti-inflammatory drug therapy attenuated the toxicity of the inflammation upon cholinergic cells and reduced caspases 3, 8 and 9 activity in the caudate/putamen. zVAD treatment significantly decreased the levels of caspases 3, 8 and 9 but did not provide neuroprotection for the cholinergic neurons. These results suggest that prostaglandins contribute to the degeneration of forebrain cholinergic neurons in Alzheimer's disease.

Topics: Amino Acid Chloromethyl Ketones; Animals; Anti-Inflammatory Agents, Non-Steroidal; Body Weight; Caspase Inhibitors; Choline O-Acetyltransferase; Enzyme Inhibitors; Flurbiprofen; Immunohistochemistry; Inflammation; Lipopolysaccharides; Male; Neurons; Parasympathetic Nervous System; Prosencephalon; Rats; Rats, Inbred F344