This Article Abstract Free Full Text (Free PDF) Free Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Waters, S. L. Articles by Schnellmann, R. G. Search for Related Content PubMed PubMed Citation Articles by Waters, S. L. Articles by Schnellmann, R. G. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB ANTIMYCIN A CALCIUM COMPOUNDS CALCIUM, ELEMENTAL GLYCINE MUSCIMOL STRYCHNINE Social Bookmarking                         What's this?

Examination of the Mechanisms of Action of Diverse Cytoprotectants in Renal Cell Death

Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205-7199

Rick G. Schnellmann

Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205-7199

Glycine, strychnine, muscimol, allopregnanolone, and pregnenolone sulfate act in the late phase of renal cell injury, block Cl^– influx and cell lysis induced by the mitochondrial inhibitor antimycin A, and promote the recovery of respiration and ion transport following hypoxia/reoxygenation. However, the mechanism of action of these compounds has not been completely elucidated. Recently, we have shown that calpains are critical mediators of renal cell death produced by diverse toxicants and that antimycin A exposure results in calpain translocation from the cytosol to the membrane fraction that is temporally associated with Cl^– influx and precedes cell death/lysis. The current study examined the effects of a group of diverse cytoprotectants on calpain activity and determined if calpain inhibition plays a role in the cytoprotection produced by these compounds. The cytoprotection produced by glycine, strychnine, muscimol, allopregnanolone, and pregnenolone sulfate in rabbit renal proximal tubules exposed to antimycin A was associated with the inhibition of antimycin A-induced calpain translocation. None of the cytoprotectants had a direct effect on calpain activity. All of the cytoprotectants decreased calcium-ionophore-induced cell death. Glycine, strychnine, and muscimol also blocked antimycin A mediated extracellular Ca²⁺ influx. These data suggest that the cytoprotective mechanism of action of glycine, strychnine, and muscimol involves the inhibition of antimycin A mediated extracellular Ca²⁺ influx as well as calpain translocation and associated Cl^– influx. In contrast, the mechanism of action of the neurosteroids results only from the blockade of calpain translocation and associated Cl^– influx.

Key Words: Calpains • calcium • glycine • neurosteroids • renal proximal tubules • mitochondrial dysfunction • necrosis

• 1. Aleo MD and Schnellmann RG (1992). The ncurotoxicants strychnine and bicuculline protect renal proximal tubules from mitochondrial inhibitor induced cell death. Life Sci. 51: 1783–1787.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 2. Baines AD, Shaikh N, and Ho P (1990). Mechanism of perfused kidney cytoprotection by alanine and glycine. Am. J. Physiol. 259: F80–F87.[Web of Science][Medline] [Order article via Infotrieve]
• 3. Bronk SF and Gores GJ (1993). pH-Dependent nonlysosomal proteolysis contributes to lethal anoxic injury of rat hepatocytes. Am. J. Physiol. 264: G744–G751.[Web of Science][Medline] [Order article via Infotrieve]
• 4. Edelstein CL, Wieder ED, Gengaro PE, Burke TJ, and Schrier RW (1994). Hypoxia-induced calpain activation in rat proximal tubules. J. Am. Soc. Nephrol. 5: 896.
• 5. Edelstein CL, Wieder ED, Yaqoob MM, Gengaro PE, Burke TJ, Nemenoff RA, and Schrier RW (1995). The role of cysteine proteases in hypoxia-induced rat renal proximal tubular injury. Proc. Natl. Acad. Sci. USA 92: 7662–7666.[Abstract/Free Full Text]
• 6. Gornall AG, Bardawill CJ, and David MM (1949). Determination of serum proteins by means of the biuret reaction. J. Biol. Chem. 177: 751–766.[Free Full Text]
• 7. Heyman SN, Brezis M, Epstein FH, Spokes K, and Rosen S (1992). Effect of glycine and hypertrophy on renal outer medullary hypoxic injury in ischemia reflow and contrast nephropathy. Am. J. Kidney Dis. 19: 578–568.[Web of Science][Medline] [Order article via Infotrieve]
• 8. Heyman SM, Rosen S, Silva P, Spokes K, Egorin MJ, and Epstein FH (1991). Protective action of glycine in cisplatin nephrotoxicity. Kidney Int. 40: 273–279.[Web of Science][Medline] [Order article via Infotrieve]
• 9. Li Q, Bowmer CJ, and Yates MS (1995). Amelioration of cisplatin nephrotoxicity with glycine: Dose dependency in rats. J. Pharm. Pharmacol. 47: 223–226.[Web of Science][Medline] [Order article via Infotrieve]
• 10. Lowry OJ, Rosebrough AL, Farr AL, and Randall RJ (1951). Protein measurement with folin phenol reagent. J. Biol. Chem. 193: 265–275.[Free Full Text]
• 11. Mangino MJ, Murphy MK, Grabau GG, and Anderson CB (1991). Protective effects of glycine during hypothermic renal ischemia-reperfusion injury. Am. J. Physiol. 261: F841–F848.[Web of Science][Medline] [Order article via Infotrieve]
• 12. Marsh DC, Hjelmhaug PK, Vreugdenhill PK, Belzer FO, and Southard JH (1991). Glycine prevention of cold ischemic injury in isolated hepatocytes. Cryobiology 28: 105–109.[CrossRef][Medline] [Order article via Infotrieve]
• 13. Miller GW, Lock EA, and Schnellmann RG (1994). Strychnine and glycine protect renal proximal tubules from various nephrotoxicants and act in the late phase of necrotic cell injury. Toxicol. Appl. Pharmacol. 125: 192–197.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 14. Miller GW and Schnellmann RG (1993). Cytoprotection by inhibition of chloride channels: The mechanism of action of glycine and strychnine. Life Sci. 53: 1211–1215.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 15. Miller GW and Schnellmann RG (1995). Inhibitors of renal chloride transport do not block toxicant-induced chloride influx in the proximal tubule. Toxicol. Lett. 76: 179–184.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 16. Moran JH and Schnellmann RG (1996). A rapid β-NADH-linked fluorescence assay for lactate dehydrogenase in cellular death. J. Pharmacol. Toxicol. Methods 36: 41–44.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 17. Moran JH and Schnellmann RG (1997). Diverse cytoprotectants prevent cell lysis and promote recovery of respiration and ion transport. Biochem. Biophys. Res. Commun. 234: 275–277.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 18. Nichols JC, Bronk SF, Mellgren RL, and Gores GJ (1994). Inhibition of nonlysosomal calcium-dependent proteolysis by glycine during anoxic injury of rat hepatocytes. Gastroenterology 106: 168–176.[Web of Science][Medline] [Order article via Infotrieve]
• 19. Nissim I and Weinberg JM (1996). Glycine attenuates Fanconi syndrome induced by maleate or ifosfamide in rats. Kidney Int. 49: 684–695.[Web of Science][Medline] [Order article via Infotrieve]
• 20. Rodeheaver DP, Aleo MD, and Schnellmann RG (1990). Differences in enzymatic and mechanical isolated rabbit renal proximal tubules: Comparison in long-term incubation. In Vitro Cell. Dev. Biol. 26: 898–904.[Web of Science][Medline] [Order article via Infotrieve]
• 21. Schilling MK, den Butter G, Saunder A, Lindell S, Belzer FO, and Southard JH (1991). Membrane stabilizing effects of glycine during kidney cold storage and reperfusion. Transplant. Proc. 23: 2387–2389.[Web of Science][Medline] [Order article via Infotrieve]
• 22. Schnellmann RG (1991). Pathophysiology of nephrotoxic cell injury. In: Diseases of the Kidney, Vol. 2, RW Schrier and CW Gottschalk (eds). Little, Brown, and Company, Boston, pp. 1049–1068.
• 23. Silva P, Rosen S, Spokes K, and Epstein FH (1991). Effect of glycine on medullary thick ascending limb injury in perfused kidneys. Kidney Int. 39: 653–658.[Web of Science][Medline] [Order article via Infotrieve]
• 24. Varani J, Ginsburg I, Johnson KJ, Gibbs DF, Weinberg JM, and Ward PA (1991). Amino acids and metal ions protect endothelial cells from lethal injury. Fed. Am. Soc. Exp. Biol. J. 5: 887.
• 25. Venkatachalam MA, Weinberg JM, Patel Y, Saikumar P, and Dong Z (1996). Cytoprotection of kidney epithelial cells by compounds that target amino acid gated chloride channels. Kidney Int. 49: 449–460.[Web of Science][Medline] [Order article via Infotrieve]
• 26. Waters SL, Miller GW, and Schnellmann RG (1997). Neurosteroid modulation of cell death. Am. J. Physiol. 273: F869–F876.[Web of Science][Medline] [Order article via Infotrieve]
• 27. Waters SL, Sarang SS, Wang KKW, and Schnellmann RG (1997). Calpains mediate calcium and chloride influx during the late phase of cell injury. J. Pharmacol. Exp. Ther. 283: 1177–1184.[Abstract/Free Full Text]
• 28. Waters SL and Schnellmann RG (1996). Extracellular acidosis and chloride channel inhibitors act in the late phase of cellular injury to prevent death. J. Pharmacol. Exp. Ther. 278: 1012–1017.[Abstract/Free Full Text]
• 29. Weinberg JM, Davis JA, Roeser NF, and Venkatachalam MA (1991). Role of increased cytosolic free calcium in the pathogenesis of rabbit proximal tubule cell injury and protection by glycine or acidosis. J. Clin. Invest. 87: 581–590.[Web of Science][Medline] [Order article via Infotrieve]
• 30. Weinberg JM, Varani J, Johnson KJ, Roeser NF, Dame MK, Davis JA, and Venkatachalam MA (1992). Protection of human umbilical vein endothelial cells by glycine and structurally related amino acids against calcium and hydrogen peroxide-induced lethal cell injury. Am. J. Pathol. 140: 457–471.[Abstract]
• 31. Weinberg JM, Venkatachalam MA, Garzo-Quintero R, Roeser NF, and Davis JA (1990). Structural requirements for protection by small amino acids against hypoxic injury in kidney proximal tubules. Fed. Am. Soc. Exp. Biol. J. 4: 3347–3354.
• 32. Zhong A, Jones S, and Thurman RG (1996). Glycine minimizes reperfusion injury in a low-flow reflow liver perfusion model in the rat. Am. J. Physiol. 270: G332–G338.[Web of Science][Medline] [Order article via Infotrieve]

Toxicologic Pathology, Vol. 26, No. 1, 58-63 (1998)
DOI: 10.1177/019262339802600108


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				X. Liu, J. J. Rainey, J. F. Harriman, and R. G. Schnellmann Calpains mediate acute renal cell death: role of autolysis and translocation Am J Physiol Renal Physiol, October 1, 2001; 281(4): F728 - F738. [Abstract] [Full Text] [PDF]

				Z. Zhong, G. E. Arteel, H. D. Connor, M. Yin, M. V. Frankenberg, R. F. Stachlewitz, J. A. Raleigh, R. P. Mason, and R. G. Thurman Cyclosporin A increases hypoxia and free radical production in rat kidneys: prevention by dietary glycine Am J Physiol Renal Physiol, October 1, 1998; 275(4): F595 - F604. [Abstract] [Full Text] [PDF]

This Article Abstract Free Full Text (Free PDF) Free Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Waters, S. L. Articles by Schnellmann, R. G. Search for Related Content PubMed PubMed Citation Articles by Waters, S. L. Articles by Schnellmann, R. G. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB ANTIMYCIN A CALCIUM COMPOUNDS CALCIUM, ELEMENTAL GLYCINE MUSCIMOL STRYCHNINE Social Bookmarking                         What's this?