Hyperosmolality impairs ammonia-mediated inflammation: implications for the renal medulla.
Although ammonia modifies the third component of complement (C3) and activates the alternative pathway, inflammation is not seen in the renal medulla where ammonia concentrations are normally elevated. We examined the effect of the unique hyperosmolar milieu of the renal medulla on the interaction of ammonia with C3 and the capacity of ammonia-modified C3 (NH3.C3) to induce cytolytic injury and stimulate neutrophils (PMN). Incubation of purified human C3 with ammonia in concentrations found in urine results in significant disruption of the C3 thiolester bond compared with ammonia-free controls. Coincubation with urinary osmolytes and hyperosmolar NaCl and urea does not impair thiolester disruption over a range of ammonia concentrations. However, hyperosmolar NaCl and urea virtually abolish cytolytic injury mediated by the alternative pathway. Coincubation with the organic osmolytes betaine, sorbitol, and inositol fails to reverse this inhibitory effect of hyperosmolar NaCl and urea. Hyperosmolar NaCl and urea also suppress lytic injury mediated by ammonia and complement in MDCK, a cell line derived from canine distal tubular epithelium. Both PMN degranulation and respiratory burst responses to NH3.C3 are significantly blunted in the presence of hyperosmolar NaCl and urea. Hyperosmolality also impairs PMN responses to the formyl peptide N-formyl-Met-Leu-Phe and phorbol 12-myristate 13-acetate (PMA). Therefore, in an in vitro setting of hyperosmolar NaCl and urea, amidation of C3 occurs, but subsequent membrane-directed and receptor-mediated functions of NH3.C3 are markedly impaired.(ABSTRACT TRUNCATED AT 250 WORDS)
Clark
E C
EC
Department of Medicine, University of Rochester School of Medicine, New York 14621.
Nath
K A
KA
Hostetter
T H
TH
Hostetter
M K
MK
D000641:Ammonia / Q000494:pharmacology; D000818:Animals; D015550:Cell Degranulation / Q000187:drug effects; D003176:Complement C3 / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007249:Inflammation / Q000139:chemically induced*; D007679:Kidney Medulla / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology; D009994:Osmolar Concentration; D012965:Sodium Chloride / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology
D003176:Complement C3; D012965:Sodium Chloride; D000641:Ammonia; D014508:Urea