JOURNAL
OF SURGICAL
RESEARCH
Complement
%,
74-78 (1979)
Activation
Early in Endotoxin
Shock
MATS HEIDEMAN, M.D.,’ BERTIL KAIJSER, M.D., AND LARS-ERIK GELIN, M.D. Department
of Surgery I and Institute of Medical Microbiology, Gothenburg, Sweden
University
of Gothenburg,
Submitted for publication March 14, 1978 The effect of endotoxin infusion was studied in dogs. Aggregation of platelets in the blood and trapping in the lung were observed. Endotoxin infusion was initially followed by leucopenia. Hemolysis of red cells corresponding to about 1% of the erythrocytes was observed. A blood pressure decrease and a tracheal insufflation pressure increase followed injury. Impaired oxygenation of arterial blood was registered. The infusion was accompanied by complement activation. Its relation to the phenomena studied is discussed.
Septic shock is known to induce blood pressure instability, thrombocytopenia, leucopenia, and respiratory impairment [5, 181. The similarity between anaphylactic and endotoxin shock and the effect of histamine infusion has been pointed out [5, 17, 181. Both in viva [2, 181and in vitro studies [ 1,4, 81 have revealed a complement activation by endotoxin. The aim of this study was to investigate the relationship between early hematological, circulatory, and respiratory reactions and changes of complement activity in serum after intravenous infusion of endotoxin. MATERIAL
AND METHODS
intravenous infusion of 2 ml of saline/kg body weightihr was given throughout the experiments. No heparin was given. In seven dogs, a thoracotomy on the right side was carried out for repeated lung biopsies. In two dogs, the splenic arteries and veins were ligated to evaluate the influence of the spleen on cell behavior. Endotoxin shock was induced by intravenous infusion of lipopoly saccharide W, extracted from E. coli (Difco Laboratories, Detroit, Mich. U. S. A.). Two milligrams per kilogram of body weight was infused for 5 min, leading to a mortality of more than 50% within the first 6 hr after infusion. Three additional dogs were used for control. They were anesthetized, catheterized, and followed by repeated measurements of the same tissue and blood parameters as the 14 experimental dogs for 7 to 10 hr. At the end of the experiments, the animals were killed, and autopsies performed. The methods used for determination of hematological, hemodynamic, and respiratory variables have been described in a previous paper [7]. Titration of whole complement in serum was performed according to the method described by Mayer [9].
Fourteen mongrel dogs of either sex weighing 16 to 30 kg were used. They were anesthetized with Ketalar and given Pavulon for muscle relaxation. After endotracheal intubation, the dogs were connected to an Engstrom respirator for controlled respiration with ambient air. The volume and rate were kept constant during the experiments except that the dogs were hyperinflated twice an hour to prevent atelectasis. One polyethylene catheter was introduced into the carotid artery and another into the jugular vein with its tip at the right atrium. An Statistical * Address requests for reprints to: Mats Heideman, M.D., Theological Lab, Dept. of Surgery I, Sahlgrenska sjukhuset, S-413 45 Gothenburg, Sweden. 0022~4804/79/010074-05$01.00/0 CopyriSht AU rights
Q 1979 by Academic Press, Inc. of reproduction in any fom reserved.
Evaluations
Means and standard errors of the mean were calculated by standard methods. Fish74
HEIDEMAN
ET AL.: COMPLEMENT
ACTIVATION
IN ENDOTOXIN
75
SHOCK
TABLE 1 CHANGES
IN
BLOOD
Leucocytes (x 1OVliter)
Phlt&tS ( x IOViter) Control O-O.5 hr l-2 hr 3-4 hr S-6 hr after infusion
I86 64 106 130
? 16 (14) 2 8 (14)*** t I4 (14)*** f I5 (lo)*
6ooO I400 1900 3900
Ill
2 IO (6)**
8700 k 2300
a Mean values f SEM. Figures in parentheses after infusion. Significant differences are indicated * P 4 0.05. **P < 0.01. ***FJ c 0.001
AFTER INFUSION
z 700 z 300 f 300 2 1200
(14) (I,)*** (14)*** (IO) (6)
OF ENDOTOXIN”
Hemolysis (absorbance at 541 nm) 0.238 0.374 0.619 0.546
c 2 2 k
er’s permutation test was used for comparisons of initial values with the values obtained at defined time intervals after infusion. A P value
OF SURGICAL
RESEARCH
Complement
%,
74-78 (1979)
Activation
Early in Endotoxin
Shock
MATS HEIDEMAN, M.D.,’ BERTIL KAIJSER, M.D., AND LARS-ERIK GELIN, M.D. Department
of Surgery I and Institute of Medical Microbiology, Gothenburg, Sweden
University
of Gothenburg,
Submitted for publication March 14, 1978 The effect of endotoxin infusion was studied in dogs. Aggregation of platelets in the blood and trapping in the lung were observed. Endotoxin infusion was initially followed by leucopenia. Hemolysis of red cells corresponding to about 1% of the erythrocytes was observed. A blood pressure decrease and a tracheal insufflation pressure increase followed injury. Impaired oxygenation of arterial blood was registered. The infusion was accompanied by complement activation. Its relation to the phenomena studied is discussed.
Septic shock is known to induce blood pressure instability, thrombocytopenia, leucopenia, and respiratory impairment [5, 181. The similarity between anaphylactic and endotoxin shock and the effect of histamine infusion has been pointed out [5, 17, 181. Both in viva [2, 181and in vitro studies [ 1,4, 81 have revealed a complement activation by endotoxin. The aim of this study was to investigate the relationship between early hematological, circulatory, and respiratory reactions and changes of complement activity in serum after intravenous infusion of endotoxin. MATERIAL
AND METHODS
intravenous infusion of 2 ml of saline/kg body weightihr was given throughout the experiments. No heparin was given. In seven dogs, a thoracotomy on the right side was carried out for repeated lung biopsies. In two dogs, the splenic arteries and veins were ligated to evaluate the influence of the spleen on cell behavior. Endotoxin shock was induced by intravenous infusion of lipopoly saccharide W, extracted from E. coli (Difco Laboratories, Detroit, Mich. U. S. A.). Two milligrams per kilogram of body weight was infused for 5 min, leading to a mortality of more than 50% within the first 6 hr after infusion. Three additional dogs were used for control. They were anesthetized, catheterized, and followed by repeated measurements of the same tissue and blood parameters as the 14 experimental dogs for 7 to 10 hr. At the end of the experiments, the animals were killed, and autopsies performed. The methods used for determination of hematological, hemodynamic, and respiratory variables have been described in a previous paper [7]. Titration of whole complement in serum was performed according to the method described by Mayer [9].
Fourteen mongrel dogs of either sex weighing 16 to 30 kg were used. They were anesthetized with Ketalar and given Pavulon for muscle relaxation. After endotracheal intubation, the dogs were connected to an Engstrom respirator for controlled respiration with ambient air. The volume and rate were kept constant during the experiments except that the dogs were hyperinflated twice an hour to prevent atelectasis. One polyethylene catheter was introduced into the carotid artery and another into the jugular vein with its tip at the right atrium. An Statistical * Address requests for reprints to: Mats Heideman, M.D., Theological Lab, Dept. of Surgery I, Sahlgrenska sjukhuset, S-413 45 Gothenburg, Sweden. 0022~4804/79/010074-05$01.00/0 CopyriSht AU rights
Q 1979 by Academic Press, Inc. of reproduction in any fom reserved.
Evaluations
Means and standard errors of the mean were calculated by standard methods. Fish74
HEIDEMAN
ET AL.: COMPLEMENT
ACTIVATION
IN ENDOTOXIN
75
SHOCK
TABLE 1 CHANGES
IN
BLOOD
Leucocytes (x 1OVliter)
Phlt&tS ( x IOViter) Control O-O.5 hr l-2 hr 3-4 hr S-6 hr after infusion
I86 64 106 130
? 16 (14) 2 8 (14)*** t I4 (14)*** f I5 (lo)*
6ooO I400 1900 3900
Ill
2 IO (6)**
8700 k 2300
a Mean values f SEM. Figures in parentheses after infusion. Significant differences are indicated * P 4 0.05. **P < 0.01. ***FJ c 0.001
AFTER INFUSION
z 700 z 300 f 300 2 1200
(14) (I,)*** (14)*** (IO) (6)
OF ENDOTOXIN”
Hemolysis (absorbance at 541 nm) 0.238 0.374 0.619 0.546
c 2 2 k
er’s permutation test was used for comparisons of initial values with the values obtained at defined time intervals after infusion. A P value
