phalloidine and Kidney-Diseases

The mitochondria are a critical target for cisplatin-associated nephrotoxicity. Though nitric oxide formation has been implicated in the toxicity of cisplatin, this formation has not so far been related to a possible activation of mitochondrial nitric oxide synthase (mNOS). We show here that the upregulation of oxide mNOS and peroxynitrite formation in cisplatin treatment are key events that influence the development of the harmful parameters described in cisplatin-associated kidney failure. We confirm this by isolating the mitochondrial fraction of the kidney and across different access routes such as the use of a specific inhibitor of neuronal NOS, L-NPA, a peroxynitrite scavenger, FeTMPyP, and a peroxynitrite donor, SIN-1. The in vitro studies corroborated the information obtained in the in vivo experiments. The administration of cisplatin reveals a clear upregulation in the transcription of neuronal NOS and an increase in the levels of nitrites in the mitochondrial fractions of the kidneys. The upregulated transcription directly affects the cytoskeleton structure and the apoptosis. The inhibition of neuronal NOS reduces the levels of nitrites, cell death, and cytoskeleton derangement. Peroxynitrite is involved in the mechanism promoting the NOS transcription. In addition, in controls SIN-1 imitates the effects of cisplatin. In summary, we demonstrate that upregulation of mNOS in cisplatin treatment is a key component in both the initiation and the spread of cisplatin-associated damage in the kidney. Furthermore, peroxynitrite formation is directly involved in this process.

Topics: Animals; Antineoplastic Agents; Apoptosis; Blotting, Western; Caspase 3; Cisplatin; Coloring Agents; Cytochromes c; Cytoskeleton; Immunohistochemistry; Kidney Diseases; Male; Mitochondria; Molsidomine; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Peroxynitrous Acid; Phalloidine; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Interleukin 2 (IL-2) is a potent cytokine with diverse effects, including the ability to stimulate lymphocyte differentiation into cells capable of lysing tumor. Its therapeutic efficacy is limited because of side effects such as breakdown of the microvascular barrier and edema. Control of the microvascular barrier is in part regulated by endothelial cell cytoskeletal contractile proteins. This study tests whether the cyclopeptides that maintain actin filament organization and distribution and reduce macromolecular flux across the endothelial cell junction in vitro would similarly maintain barrier tightness and prevent early edema produced by IL-2 in vivo. Anesthetized rats were treated at 30-min periods with intravenous saline (0.5 ml, n = 41), phalloidin (20 micrograms in 0.5 ml, n = 21), or antamanide, (20 micrograms in 0.5 ml, n = 21), starting 30 min before the 1-h infusion of 10(6) U of recombinant human IL-2 or saline. Six hours after the start of IL-2, there was edema in the saline/IL-2 group, as measured by increased wet-to-dry ratios (W/D) in the lungs, heart, and kidney. With saline/IL-2, bronchoalveolar lavage (BAL) fluid contained an elevated protein concentration and higher plasma thromboxane levels compared with controls. The number of neutrophils sequestered in the lungs was more than twice that of saline controls. Phalloidin significantly attenuated edema in lung and reduced BAL protein leak. Antamanide treatment was as effective in limiting lung and heart edema, but, in contrast to phalloidin, antamanide prevented kidney edema and did not lead to an alteration in the liver W/D. Antamanide also prevented BAL fluid protein leak.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Capillary Permeability; Chemical and Drug Induced Liver Injury; Edema; Edema, Cardiac; Interleukin-2; Kidney Diseases; Liver Diseases; Male; Peptides, Cyclic; Phalloidine; Pulmonary Edema; Rats; Rats, Inbred Strains; Thromboxane B2

Plasma protein profiles were investigated electrophoretically in rats with aminonucleoside-induced glomerular damage, uranyl-nitrate-induced tubular damage, phalloidin-induced hepatocyte membrane damage, adjuvant arthritis as a model of chronic inflammation, and phenyl isothiocyanate pleurisy as a model of acute inflammation. Characteristic profiles were observed for each experimental disease, and their development and seriousness could be evaluated. Similar studies are already routinely used in clinical investigations of human plasma.

Topics: Animals; Blood Proteins; Chemical and Drug Induced Liver Injury; Electrophoresis, Cellulose Acetate; Isothiocyanates; Kidney Diseases; Liver Diseases; Phalloidine; Pleurisy; Puromycin Aminonucleoside; Rats; Rodent Diseases; Thiocyanates; Uranyl Nitrate