Does endotoxin-activated complement alter myocellular sodium homeostasis during sepsis?

Weiyang Wang, Ken Okamoto, Danny O. Jacobs

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: Inappropriate complement activation is closely related to tissue injury and organ dysfunction during systemic infection. It is not clear, however, if endotoxin-induced complement activation is responsible for changes in myocellular sodium homeostasis during sepsis. Methods: Rats underwent cecal ligation and puncture (CLP) or sham operation. Twenty-four hours after operation, fast-twitch extensor digitorum longus (EDL) muscles were isolated, incubated at 30°C for 1 hour in Krebs-Henseleit buffer (KHB) (pH 7.4), and used to measure intracellular Na+ and K+ contents. Blood samples were collected to measure serum hemolytic complement activity and endotoxin levels. In addition, EDL muscles isolated from normal animals were incubated at 30°C for 1 hour with zymosan-activated (10 mg/mL at 37°C for 1 hour) rat sera, with lipopolysaccharide (LPS)-activated (LPS from Escherichia coli 055: B5, 10 or 200 μg/mL at 37°C for 30 minutes) rat sera, with heat-inactivated (56°C for 30 minutes) rat sera, with LPS (1 or 20 μg/mL), or in KHB. EDL muscles isolated from normal animals were also incubated with septic sera collected 6 or 24 hours after CLP with or without administration of soluble complement receptor type 1 (20 mg/kg, intraperitoneally). Myocellular Na+ and K+ contents ([Na+]i and [K+]i) were assayed using "washout" technique. Soluble C5b-9 complex levels in zymosan-activated or LPS-activated human sera were determined by enzyme-linked immunosorbent assay to evaluate the degree of complement activation induced by zymosan or LPS. Results: Myocellular [Na+]i and [Na+]i/[K+]i ratios increased significantly 24 hours after CLP as compared with sham operation and were associated with decreased serum hemolytic complement activity and increased serum endotoxin levels. Zymosan-activated rat sera at sublytic concentrations markedly increased [Na+]i and [Na+]i/[K+]i ratios in isolated EDL muscles relative to heat-inactivated rat sera. LPS-activated rat sera, however, did not alter these two indices. In addition, myocellular [Na+]i and [Na+]i/[K+]i ratios were equivalent among normal EDL muscles incubated with septic sera, soluble complement receptor type 1-treated septic sera, or KHB. Conclusion: These results collectively suggest that polymicrobial sepsis, as produced by CLP, alters sodium homeostasis in fast-twitch skeletal muscles in association with changes in systemic complement activation and circulating endotoxin levels. Although endotoxin can activate the complement cascade, endotoxininduced complement activation does not appear to be responsible for changes in myocellular sodium homeostasis observed during sepsis in rats.

Original languageEnglish (US)
Pages (from-to)951-961
Number of pages11
JournalJournal of Trauma - Injury, Infection and Critical Care
Volume52
Issue number5
StatePublished - 2002
Externally publishedYes

Fingerprint

Endotoxins
Sepsis
Homeostasis
Sodium
Serum
Complement Activation
Lipopolysaccharides
Zymosan
Punctures
Ligation
Muscles
Complement Receptors
Complement System Proteins
Hot Temperature
Skeletal Muscle
Enzyme-Linked Immunosorbent Assay
Escherichia coli

Keywords

  • Complement activation
  • Complement membrane attack complex
  • Critical care
  • Lipopolysaccharide
  • Potassium
  • Septic shock
  • Skeletal muscle
  • Sodium
  • Zymosan

ASJC Scopus subject areas

  • Surgery

Cite this

Does endotoxin-activated complement alter myocellular sodium homeostasis during sepsis? / Wang, Weiyang; Okamoto, Ken; Jacobs, Danny O.

In: Journal of Trauma - Injury, Infection and Critical Care, Vol. 52, No. 5, 2002, p. 951-961.

Research output: Contribution to journalArticle

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T1 - Does endotoxin-activated complement alter myocellular sodium homeostasis during sepsis?

AU - Wang, Weiyang

AU - Okamoto, Ken

AU - Jacobs, Danny O.

PY - 2002

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N2 - Background: Inappropriate complement activation is closely related to tissue injury and organ dysfunction during systemic infection. It is not clear, however, if endotoxin-induced complement activation is responsible for changes in myocellular sodium homeostasis during sepsis. Methods: Rats underwent cecal ligation and puncture (CLP) or sham operation. Twenty-four hours after operation, fast-twitch extensor digitorum longus (EDL) muscles were isolated, incubated at 30°C for 1 hour in Krebs-Henseleit buffer (KHB) (pH 7.4), and used to measure intracellular Na+ and K+ contents. Blood samples were collected to measure serum hemolytic complement activity and endotoxin levels. In addition, EDL muscles isolated from normal animals were incubated at 30°C for 1 hour with zymosan-activated (10 mg/mL at 37°C for 1 hour) rat sera, with lipopolysaccharide (LPS)-activated (LPS from Escherichia coli 055: B5, 10 or 200 μg/mL at 37°C for 30 minutes) rat sera, with heat-inactivated (56°C for 30 minutes) rat sera, with LPS (1 or 20 μg/mL), or in KHB. EDL muscles isolated from normal animals were also incubated with septic sera collected 6 or 24 hours after CLP with or without administration of soluble complement receptor type 1 (20 mg/kg, intraperitoneally). Myocellular Na+ and K+ contents ([Na+]i and [K+]i) were assayed using "washout" technique. Soluble C5b-9 complex levels in zymosan-activated or LPS-activated human sera were determined by enzyme-linked immunosorbent assay to evaluate the degree of complement activation induced by zymosan or LPS. Results: Myocellular [Na+]i and [Na+]i/[K+]i ratios increased significantly 24 hours after CLP as compared with sham operation and were associated with decreased serum hemolytic complement activity and increased serum endotoxin levels. Zymosan-activated rat sera at sublytic concentrations markedly increased [Na+]i and [Na+]i/[K+]i ratios in isolated EDL muscles relative to heat-inactivated rat sera. LPS-activated rat sera, however, did not alter these two indices. In addition, myocellular [Na+]i and [Na+]i/[K+]i ratios were equivalent among normal EDL muscles incubated with septic sera, soluble complement receptor type 1-treated septic sera, or KHB. Conclusion: These results collectively suggest that polymicrobial sepsis, as produced by CLP, alters sodium homeostasis in fast-twitch skeletal muscles in association with changes in systemic complement activation and circulating endotoxin levels. Although endotoxin can activate the complement cascade, endotoxininduced complement activation does not appear to be responsible for changes in myocellular sodium homeostasis observed during sepsis in rats.

AB - Background: Inappropriate complement activation is closely related to tissue injury and organ dysfunction during systemic infection. It is not clear, however, if endotoxin-induced complement activation is responsible for changes in myocellular sodium homeostasis during sepsis. Methods: Rats underwent cecal ligation and puncture (CLP) or sham operation. Twenty-four hours after operation, fast-twitch extensor digitorum longus (EDL) muscles were isolated, incubated at 30°C for 1 hour in Krebs-Henseleit buffer (KHB) (pH 7.4), and used to measure intracellular Na+ and K+ contents. Blood samples were collected to measure serum hemolytic complement activity and endotoxin levels. In addition, EDL muscles isolated from normal animals were incubated at 30°C for 1 hour with zymosan-activated (10 mg/mL at 37°C for 1 hour) rat sera, with lipopolysaccharide (LPS)-activated (LPS from Escherichia coli 055: B5, 10 or 200 μg/mL at 37°C for 30 minutes) rat sera, with heat-inactivated (56°C for 30 minutes) rat sera, with LPS (1 or 20 μg/mL), or in KHB. EDL muscles isolated from normal animals were also incubated with septic sera collected 6 or 24 hours after CLP with or without administration of soluble complement receptor type 1 (20 mg/kg, intraperitoneally). Myocellular Na+ and K+ contents ([Na+]i and [K+]i) were assayed using "washout" technique. Soluble C5b-9 complex levels in zymosan-activated or LPS-activated human sera were determined by enzyme-linked immunosorbent assay to evaluate the degree of complement activation induced by zymosan or LPS. Results: Myocellular [Na+]i and [Na+]i/[K+]i ratios increased significantly 24 hours after CLP as compared with sham operation and were associated with decreased serum hemolytic complement activity and increased serum endotoxin levels. Zymosan-activated rat sera at sublytic concentrations markedly increased [Na+]i and [Na+]i/[K+]i ratios in isolated EDL muscles relative to heat-inactivated rat sera. LPS-activated rat sera, however, did not alter these two indices. In addition, myocellular [Na+]i and [Na+]i/[K+]i ratios were equivalent among normal EDL muscles incubated with septic sera, soluble complement receptor type 1-treated septic sera, or KHB. Conclusion: These results collectively suggest that polymicrobial sepsis, as produced by CLP, alters sodium homeostasis in fast-twitch skeletal muscles in association with changes in systemic complement activation and circulating endotoxin levels. Although endotoxin can activate the complement cascade, endotoxininduced complement activation does not appear to be responsible for changes in myocellular sodium homeostasis observed during sepsis in rats.

KW - Complement activation

KW - Complement membrane attack complex

KW - Critical care

KW - Lipopolysaccharide

KW - Potassium

KW - Septic shock

KW - Skeletal muscle

KW - Sodium

KW - Zymosan

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