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Plasma prokallikrein and kininogens in burned patients
THROMBOSIS RESEARCH 41; 537-543, 1986 0049-3848/86 $3.00 t .OO Printed in the USA. Copyright (c) 1986 Pergamon Press Ltd. All rights reserved.
PLASMA PROKALLIKREIN
AND KININOCENS IN BURNED PATIENTS
Adam(l,2), Jac ues Damas(3), Adelin Albertt4), Pierre E&l), Jacques Marichit 3 1, Ciermain Calayqll, Philippe Laurent(6). l- Laboratoire de Biologie clinique, C.H. Sainte-Ode, B-6970 Baconfoy, 2Laboratoire de Radioimmunologie, 3- Service de Physiologie, 4- Laboratoire de Chimie mkdicale, Universite de Liege, B-4020 Liege, Belgique. 5- Centre des BrOlCs, H6pital E. Herriot, 6- Laboratoire de Physiopathologie de 1’Inflammation, Institut Pasteur, Lyon, France. Albert
(Received 27.6.1985; Accepted in revised form 15.10.1985 by Editor B. Vargaftig) (Received in final form by Executive Editorial Office 26.11.1985)
ABSTRACT Using specific immunological (1) and enzymatic (2) methods, we have measured prokallikrein, total, high, and low molecular weight kininogens in 36 severely burned patients. At admission to the intensive care unit, all constituents were significantly decreased when compared to previously defined reference intervals. The values remained low during the three first days after burn. The changes affecting total and low molecular weight kininogens were significantly correlated (p < 0.05) with the severity of the burn area. Prokallikrein and kininogens levels were also closely related to the concentrations of C3c and C4 complement factors.
INTRODUCTION Rocha e Silva and Antonio (31 have observed that immersing rat paw in hot water induced the appearance of kinins in the perfusate of the subcutaneous spaces. Similar results has been obtained by others (4, 51, whereas Edery and Lewis (6) have detected a kinin-forming activity in the lymph draining the burned limb of the dog (6). Kinins are released from low and high molecular weight kininogens (LMW- and respectively) by several specific (e.g., plasma and tissue HMW kininogens, kallikreins) or non specific (e.g., trypsin, leucocytic cathepsin) enzymes. These polypeptides have proinflammatory actions and might be involved in the inflammatory response to the thermal injury. The contribution of the kinin system to inflammatory processes has been extensively investigated in animal, but until now very few results have been reported in man (71. Key words : prokallikrein, inhibitor, burned patients.
LMW- and HMW kininogens, a2 537
cystein
proteinase
538
KININ SYSTEM IN BURN
Vol.
41,
No. 4
Using recently developed specific immunological and enzymatic methods (1, 21, of prokallikrein and kininogens in we quantified the plasma concentration severely burned patients at the admission to intensive care unit and during the first three days of the acute hospitalisation. We studied the evolution of the same constituents according to outcome, death or survival, and correlated the observations with the severity of the burn and with the complement factors. We argue that the changes affecting the kinin precursors (kininogensl and their specific plasma activator (prokallikrein) can inform us about their involvement in the inflammatory response pathway to skin burn.
iMATERIAL --
AND
METHODS
Patients. 36 burned patients (20 men and 16 women, 24 f- 19 years) -admitted to the “Centre des BrQICs, H8pital E. Herriot”, Lyon, France, have been studied. The patients were classified according to the extent of the burn, assessed by both the measurements of Burn Skin Area (BSA) and Burn Unit Skin (BUS) proposed by Sachs and Watson (81. The patients were divided into (301 and nonsurvivors (61. The two groups according to outcome : survivors cause of accidental burn and the clinical characteristics of the patients are summarized in Table I. Sampling procedure. Blood samples were collected by venipuncture at admiss= to theintensive care unit and on the two following days. After discarding the first ml to remove the “endogeneous” activator, blood was drawn in polystyrene tubes containing EDTA (final concentration : 1 mg/ml) for complement factor determinations or sodium citrate (0.1 mmol, l/IO v/v) for prokallikrein and kininogen measurements. After centrifugation (10 min.,
Cause
of the
TABLE I Characteristics of the burned burn burning flam hot fluid fluid
No of (::%, patients Severity of the burn 10 BSA (%l 2 Noof patients (6%) Benign BUS (%) 50 No of patients
Z%, 10-20 (2;%, 50-100 14 (39%)
C::%l 20-30 (II%, Heavy
patients
(n = 361
gas bursting
hot solid
(;%I
(62%)
30-40 &%1 100-150
40-50 (87y61 Severe > 150
> 50 7 (19%)
;)Il%)
15OC, 1500 x g), the plasma was decanted with a plastic pipette and stored - 30°C. Quantification was performed within a month. Plasma total proteins were measured on an autoanalyser the biuret method. ztac-ka Works Katsuda, Japan) * Prokallikrein enzyme activity ‘was determined at 37’?gy an automated method on a Hitachi 705 autoanalyser (2); the activator was ellagic acid (Cephotest, Nyegaard, Oslo Norway) and the chromogenic substrate, PK chromozyme, commercialized by Boehringer, Mannheim (FRG). Total kininogens and HMW kininogen were quantified by radioimmunoassay (11.
KININ SYSTEM IN BURN
Vol. 41, No. 4
539
and antisera were raised in The antigens were purified in our laboratory rabbit (9). Because of the complete and equimolar cross reaction of HMW kininogen in the LMW kininogen radioimmunoassay, total kininogens can be quantified by this antiserum. The antiserum anti-HMW kininogen was immuno-adsorbed by the LMW kininogen before use and was therefore highly specific for HMW kininogen. The concentration (in mmol/L) of LMW kininogen was obtained by the difference between total kininogens and HMW the molecular weight of LMW- and HMW kininOgenS kininogen. Knowing (66.000 and 110.000, respectively) (91, the concentrations (mg/Ll of HMWLMW- and total kininogens were immediately calculated. C complement factors were quantified by laser nephelometry and C France). (H$and Neph4&ometer) using reagents purchased from Hyland (Plaisir, Statistical analysis. The mean value and standard deviation (SD) were computed for all variables. Correlations were tested by the Student t-test on where n denotes the number of observations. n-2 degrees of freedom, Response curves of the constituents of the kallikrein-kininogen system were determined for each patient by linear interpolation between the serial We compared mean response curves according to severity of measurements. the burn by applying Zerbe’s method (101. This method allows comparison of curves over the 3-day survey period, thus providing a global response assessment of group differences. In each case, an F-criterion, with degrees of freedom that depended on the observations and the period selected, was calculated.
RESULTS l-
Plasma levels of prokallikrein, kininogens and complement factors in burned patients. Upon admission to the intensive care unit, prokallikrein, HMW- LMW- and total kininogens concentrations were significantly decreased (p < 0.05) when compared to previously defined reference values (1, 2) (Table II).
Mean c3c
values and
TABLE II (SD) of the kallikrein-kininogen C4 factors, at admission to Burned patients (n = 36)
Total proteins, g/L Prokallikrein, U/L Total kininogens, mg/L LMW kininogen, mg/L HMW kininogen, mg/L Cgc’ g/L C4’ g/L (a1 Significantly reduced (p <0.05). (b) See reference (11 and (2).
system constituents, intensive care unit. (al Reference values (n = 450)
52’85 (1:;; 196 (461 118 (351 78 (15) 0.69 (0.23) 0.18 (0.08)
7:: 265 175 0.; 0.35
(b)
(1:;; (411 (36) (1&6) (0.07)
Prokallikrein, total and LMW kininogens were markedly diminished: 26, and 33 respectively. These changes important than protein losses $5%); therefore the relative conc~~~~ati~~e(mg/g proteins) of total and LMW kininogens were also significantly reduced. The decrease in HMW kininogen values was similar to the decrease of total protein concentrations (14 %). The values of Cgc and C4 were also statistically decreased, C4 dropping by more
540
KININ SYSTEM IN BURN
Vol. 41, No. 4
than 50 percent. system during the The evolutions of the constituents of the kallikrein-kininogen in FIG. 1. The striped zone on the first three days after burn are displayed figure corresponds to reference values determined on healthy volunteers (1, 2). For each day the mean value and its standard error are represented.
60
1
Evolution of phase of the
2
prokallikrein, burn. Each
j
FIG. 1 total, HMW- and point represents the
1
LMW mean
I 2
kininogens during and its standard
3 Day
the acute error.
During the acute phase, the concentration of prokallikrein and kininogens were not significantly different from those recorded on admission. Prokallikrein, total and LIMW kininogens evolved towards the lower fence of the reference Although significantly decreased, the modif ications affecting HM W interval. and the mean values remained inside the kininogen were less pronounced, reference interval. 30 survived and 6 died from septicemia Among the 36 burned patients, We compared the response curves of average several days after admission. survivors and nonsurvivors over the 3 day observation period (FIG. 2) and found that total kininogens (F = 3.73, df = 1.35 and 39.11, p < 0.09, and LMW kininogen (F = 3.16, df 1.32 and 38.17, p < 0.07), were significantly lower in nonsurvivors. No difference however was found for prokallikrein (F = 1.66, df = 1.40 and 33.56, N.S.), and HMW kininogen (F = 2.54, df = 1.30 and 37.65, N.S.), measured in both groups of patients.
Vol. 41, No. 4
541
KININ SYSTEM IN BURN
3
120
E
100
5 F .E
80
C r 3
6o
I
1
Evolution outcome,
of prokallikrein survivors (01, and
FIG. 2 and kininogens nonsurvivors (AI.
in
2
burned
patients
3 Day
according
to
2- ------Correlation betweendasma prokallikrein and kininoge_n_s, and the extent --~------of the burn I* The correlations between the variables investigated are given in Table III. Total and LMW kininogens were negatively correlated with BSA (p < 0.05) and with the severity of the injury estimated by the BUS. No correlation however, with prokallikrein and HMW kininogen. ProkaIIikrein and observed was kininogens were positively correlated with the concentrations of C3c and C4 complement factors. TABLE III of the constituents of the kallikrein-kininogen system with the severity of the burn and the complement factors BSA BUS c4 c3c 0.27n 0.23* - 0.26 - 0.23 Prokallikrein 0.49* - 0.38X 0.56” - 0.31% Total kininogens 0.46” 0.52% - 0.38X - 0.30% LM W kininogen 0.42* 0.48* - 0.25 - 0.25 HM W kininogen (WI significant at the level p .=z0.05 Correlations at admission
542
KININ SYSTEM IN BURN
Vol.
41,
No. 4
DISCUSSION -----~ Several studies performed in animals (3-6) have shown that proteolytic enzymes and kinins are released in burned tissues. Our results obtained in man agree with these previous findings. We have shown that plasma prokallikrein and kininogens are significantly decreased after an extensive burn of the skin and remain low during the acute inflammatory response. A decreased prokallikrein activity suggests that this enzyme is activated by the injury, releasing bradykinin from HM W kininogen. The plasma concentration of HMW kininogen however is modified in the same way as total proteins. Further, values of LMW kininogen are even more decreased. This apparent discrepancy, also observed in acute pancreatitis (121, can be explained by the nature of the methods used to quantify both kininogens. We recently showed that “in vitro” the heavy chain used to quantify total and LMW kininogen was more sensitive to proteolysis than the histidin rich light chain specific of HMW kininogen (13). This observation is related to the immunogenicity of both kinds of kininogens and agrees with the results of Scott et al. (141, who observed that HMW kininogen can release bradykinin without loosing the procoagulant activity of its light chain. We also noted that the decrease in LMW kininogen was directly proportional to the severity of the burn, indicating that this kind of kininogen might be cleaved by tissue proteases released after the injury. Our findings suggest that kinins liberated early after skin burn participate in the inflammatory response. other biochemical mediators are also involved, e.g., the complement However, and C4 which are simultaneously activated in burned patients as zt%? inc3fhis stu dy. This confirms the results of Green (15) who described some differences between the exsudates induced by bradykinin or by thermal injury and our previous observation that heating induces oedema in the paw of normal rats as well as in brown Norway rats in which kinin formation does not occur (16). REFERENCES -------1.
ADAM, A., ALBERT, A., CALAY, G., CLOSSET, J., DAMAS FRANCHIMONT, P. Human kininogens of low- and high- molecular Quantification by radioirnmunoassay and determination of reference Clin. Chem. 31,- 423-426, 1985. _
2.
ADAM, A., AZZOUZI, M., BOULANGER, J., ERS, P., ALBERT, A., DAMAS. J. and FAYMONVILLE. M-E. Obtimized determination of olasma prokallikrein on a Hitachi 705’ autoanaliser. Clin. Chem. Clin. Biochem. ---------~---&3, 203-207, 1985.
3.
ROCHA e mechanism Med. Exe, --
4.
GARCIA LEME, J., HAMAMURA anti-proteases and hexadimethrine bradykinin-like substance during Pharmacol. 40, 294-309, 1970. --
5.
LIMAOS, E.A., BORGES, D.R., SOUZA-PINTO, J.C., GORDON, A.M. and PRADO, J.L. Acute turpentine inflamation and kinin release in rat-paw thermic oedema. Br. J. exp. Path. 62, 591-594, 1981.
SILVA, M. and ANTONIO, of production of a thermic 371-382, 1960. L.
A. Release of oedema (45’C)
3. and mass : values.
bradykinin and the in the rat’s paw.
and ROCHA e SILVA, M. Effect bromide on the release of heating (46°C) of rat paws. Brit.
of a J.
Vol. 41, No. 4
KININ SYSTEM IN BURN
EDERY, IM. and LEWIS, G.P. 6. lymph after tissue injury. JLasiol. _COLMAN, 7. conditions. In: Springer-Verlag,
543
Kinin forming activity 169, 568-583, 1963.
and
R.W. and WONG, P.Y. Kallikrein-kinin system Hand. eg. --- Pharmacol. --_--25 suppl. E.G. Erdijs I_-_pp 569-607, 1979.
8. SACHS, A. and WATSON, an overview. In: The immune (ed.) Baltimore: Wi~~rn~?-%%~
histamine
in
in pathologic ted.) Berlin:
T. The immune consequences of thermal injury: cons3uences of thermal in&. J. Ninnemann _1-----ppT%, 1981.
ADAM, A., CLOSSET, J. and DAMAS, J. 4 convenient and simultaneous 9. purification of high and low molecular weight human kininogens. Mol. Physiol. -I, 177-184, 1985. 10. ZERBE, extended to 1979.
G.O. Randomization growth and response
11. ARMSTRONG, D. In: Springer-Verlag, pp 434-481,
analysis curves.
of the completely randomized design J. Assoc., 74, 215-221, -----_-Am. Stat.
Pharmacol. Hand. exp, -_-_I---1970.
LASSON,A. and OHLSSON, 12. during acute pancreatitis in man.
K. Changes Thromb. Res. -_
25.
E.G.
Erdbs
in the kallikrein 35, 27-41, 1984.
13. ADAM, A., DAMAS, J., ERS, P., ALBERT, A., Signif ication physiopathologique des dosages J. kininogenes humains. Pathol. Biol. I, 1986 (in press). ----
ted.1
kinin
Berlin:
system
STAS, J.L. and LECOMTE, radioimmunologiques des
14. SCOTT, C.F., SILVER, L.D., SCHAPIRA, M. and COLMAN, R.W. Cleavage of human high molecular weight kininogen markedly enhances its coagulant activity. Evidence that this molecule exists as a profactor. J. Clin. --___--__-- Invesg, 954-962, 1984. 15. GREEN, K.L. Quantitative studies on and erythrocytes in mouse paw oedema Br. _-J. ---exe, _-I_ Pathol. 59, 38-47, 1978. injury. -DAMAS, J., REMACLE-VOLON, G. 16. system in various inflammatory models 391-399, 1984.
and in
the accumulation of serum induced by bradykinin or
albumin thermal
ADAM, A. The role of the kinin the rat. Int. J. Tiss. Reac. 6 -_-__-_-___I
PLASMA PROKALLIKREIN
AND KININOCENS IN BURNED PATIENTS
Adam(l,2), Jac ues Damas(3), Adelin Albertt4), Pierre E&l), Jacques Marichit 3 1, Ciermain Calayqll, Philippe Laurent(6). l- Laboratoire de Biologie clinique, C.H. Sainte-Ode, B-6970 Baconfoy, 2Laboratoire de Radioimmunologie, 3- Service de Physiologie, 4- Laboratoire de Chimie mkdicale, Universite de Liege, B-4020 Liege, Belgique. 5- Centre des BrOlCs, H6pital E. Herriot, 6- Laboratoire de Physiopathologie de 1’Inflammation, Institut Pasteur, Lyon, France. Albert
(Received 27.6.1985; Accepted in revised form 15.10.1985 by Editor B. Vargaftig) (Received in final form by Executive Editorial Office 26.11.1985)
ABSTRACT Using specific immunological (1) and enzymatic (2) methods, we have measured prokallikrein, total, high, and low molecular weight kininogens in 36 severely burned patients. At admission to the intensive care unit, all constituents were significantly decreased when compared to previously defined reference intervals. The values remained low during the three first days after burn. The changes affecting total and low molecular weight kininogens were significantly correlated (p < 0.05) with the severity of the burn area. Prokallikrein and kininogens levels were also closely related to the concentrations of C3c and C4 complement factors.
INTRODUCTION Rocha e Silva and Antonio (31 have observed that immersing rat paw in hot water induced the appearance of kinins in the perfusate of the subcutaneous spaces. Similar results has been obtained by others (4, 51, whereas Edery and Lewis (6) have detected a kinin-forming activity in the lymph draining the burned limb of the dog (6). Kinins are released from low and high molecular weight kininogens (LMW- and respectively) by several specific (e.g., plasma and tissue HMW kininogens, kallikreins) or non specific (e.g., trypsin, leucocytic cathepsin) enzymes. These polypeptides have proinflammatory actions and might be involved in the inflammatory response to the thermal injury. The contribution of the kinin system to inflammatory processes has been extensively investigated in animal, but until now very few results have been reported in man (71. Key words : prokallikrein, inhibitor, burned patients.
LMW- and HMW kininogens, a2 537
cystein
proteinase
538
KININ SYSTEM IN BURN
Vol.
41,
No. 4
Using recently developed specific immunological and enzymatic methods (1, 21, of prokallikrein and kininogens in we quantified the plasma concentration severely burned patients at the admission to intensive care unit and during the first three days of the acute hospitalisation. We studied the evolution of the same constituents according to outcome, death or survival, and correlated the observations with the severity of the burn and with the complement factors. We argue that the changes affecting the kinin precursors (kininogensl and their specific plasma activator (prokallikrein) can inform us about their involvement in the inflammatory response pathway to skin burn.
iMATERIAL --
AND
METHODS
Patients. 36 burned patients (20 men and 16 women, 24 f- 19 years) -admitted to the “Centre des BrQICs, H8pital E. Herriot”, Lyon, France, have been studied. The patients were classified according to the extent of the burn, assessed by both the measurements of Burn Skin Area (BSA) and Burn Unit Skin (BUS) proposed by Sachs and Watson (81. The patients were divided into (301 and nonsurvivors (61. The two groups according to outcome : survivors cause of accidental burn and the clinical characteristics of the patients are summarized in Table I. Sampling procedure. Blood samples were collected by venipuncture at admiss= to theintensive care unit and on the two following days. After discarding the first ml to remove the “endogeneous” activator, blood was drawn in polystyrene tubes containing EDTA (final concentration : 1 mg/ml) for complement factor determinations or sodium citrate (0.1 mmol, l/IO v/v) for prokallikrein and kininogen measurements. After centrifugation (10 min.,
Cause
of the
TABLE I Characteristics of the burned burn burning flam hot fluid fluid
No of (::%, patients Severity of the burn 10 BSA (%l 2 Noof patients (6%) Benign BUS (%) 50 No of patients
Z%, 10-20 (2;%, 50-100 14 (39%)
C::%l 20-30 (II%, Heavy
patients
(n = 361
gas bursting
hot solid
(;%I
(62%)
30-40 &%1 100-150
40-50 (87y61 Severe > 150
> 50 7 (19%)
;)Il%)
15OC, 1500 x g), the plasma was decanted with a plastic pipette and stored - 30°C. Quantification was performed within a month. Plasma total proteins were measured on an autoanalyser the biuret method. ztac-ka Works Katsuda, Japan) * Prokallikrein enzyme activity ‘was determined at 37’?gy an automated method on a Hitachi 705 autoanalyser (2); the activator was ellagic acid (Cephotest, Nyegaard, Oslo Norway) and the chromogenic substrate, PK chromozyme, commercialized by Boehringer, Mannheim (FRG). Total kininogens and HMW kininogen were quantified by radioimmunoassay (11.
KININ SYSTEM IN BURN
Vol. 41, No. 4
539
and antisera were raised in The antigens were purified in our laboratory rabbit (9). Because of the complete and equimolar cross reaction of HMW kininogen in the LMW kininogen radioimmunoassay, total kininogens can be quantified by this antiserum. The antiserum anti-HMW kininogen was immuno-adsorbed by the LMW kininogen before use and was therefore highly specific for HMW kininogen. The concentration (in mmol/L) of LMW kininogen was obtained by the difference between total kininogens and HMW the molecular weight of LMW- and HMW kininOgenS kininogen. Knowing (66.000 and 110.000, respectively) (91, the concentrations (mg/Ll of HMWLMW- and total kininogens were immediately calculated. C complement factors were quantified by laser nephelometry and C France). (H$and Neph4&ometer) using reagents purchased from Hyland (Plaisir, Statistical analysis. The mean value and standard deviation (SD) were computed for all variables. Correlations were tested by the Student t-test on where n denotes the number of observations. n-2 degrees of freedom, Response curves of the constituents of the kallikrein-kininogen system were determined for each patient by linear interpolation between the serial We compared mean response curves according to severity of measurements. the burn by applying Zerbe’s method (101. This method allows comparison of curves over the 3-day survey period, thus providing a global response assessment of group differences. In each case, an F-criterion, with degrees of freedom that depended on the observations and the period selected, was calculated.
RESULTS l-
Plasma levels of prokallikrein, kininogens and complement factors in burned patients. Upon admission to the intensive care unit, prokallikrein, HMW- LMW- and total kininogens concentrations were significantly decreased (p < 0.05) when compared to previously defined reference values (1, 2) (Table II).
Mean c3c
values and
TABLE II (SD) of the kallikrein-kininogen C4 factors, at admission to Burned patients (n = 36)
Total proteins, g/L Prokallikrein, U/L Total kininogens, mg/L LMW kininogen, mg/L HMW kininogen, mg/L Cgc’ g/L C4’ g/L (a1 Significantly reduced (p <0.05). (b) See reference (11 and (2).
system constituents, intensive care unit. (al Reference values (n = 450)
52’85 (1:;; 196 (461 118 (351 78 (15) 0.69 (0.23) 0.18 (0.08)
7:: 265 175 0.; 0.35
(b)
(1:;; (411 (36) (1&6) (0.07)
Prokallikrein, total and LMW kininogens were markedly diminished: 26, and 33 respectively. These changes important than protein losses $5%); therefore the relative conc~~~~ati~~e(mg/g proteins) of total and LMW kininogens were also significantly reduced. The decrease in HMW kininogen values was similar to the decrease of total protein concentrations (14 %). The values of Cgc and C4 were also statistically decreased, C4 dropping by more
540
KININ SYSTEM IN BURN
Vol. 41, No. 4
than 50 percent. system during the The evolutions of the constituents of the kallikrein-kininogen in FIG. 1. The striped zone on the first three days after burn are displayed figure corresponds to reference values determined on healthy volunteers (1, 2). For each day the mean value and its standard error are represented.
60
1
Evolution of phase of the
2
prokallikrein, burn. Each
j
FIG. 1 total, HMW- and point represents the
1
LMW mean
I 2
kininogens during and its standard
3 Day
the acute error.
During the acute phase, the concentration of prokallikrein and kininogens were not significantly different from those recorded on admission. Prokallikrein, total and LIMW kininogens evolved towards the lower fence of the reference Although significantly decreased, the modif ications affecting HM W interval. and the mean values remained inside the kininogen were less pronounced, reference interval. 30 survived and 6 died from septicemia Among the 36 burned patients, We compared the response curves of average several days after admission. survivors and nonsurvivors over the 3 day observation period (FIG. 2) and found that total kininogens (F = 3.73, df = 1.35 and 39.11, p < 0.09, and LMW kininogen (F = 3.16, df 1.32 and 38.17, p < 0.07), were significantly lower in nonsurvivors. No difference however was found for prokallikrein (F = 1.66, df = 1.40 and 33.56, N.S.), and HMW kininogen (F = 2.54, df = 1.30 and 37.65, N.S.), measured in both groups of patients.
Vol. 41, No. 4
541
KININ SYSTEM IN BURN
3
120
E
100
5 F .E
80
C r 3
6o
I
1
Evolution outcome,
of prokallikrein survivors (01, and
FIG. 2 and kininogens nonsurvivors (AI.
in
2
burned
patients
3 Day
according
to
2- ------Correlation betweendasma prokallikrein and kininoge_n_s, and the extent --~------of the burn I* The correlations between the variables investigated are given in Table III. Total and LMW kininogens were negatively correlated with BSA (p < 0.05) and with the severity of the injury estimated by the BUS. No correlation however, with prokallikrein and HMW kininogen. ProkaIIikrein and observed was kininogens were positively correlated with the concentrations of C3c and C4 complement factors. TABLE III of the constituents of the kallikrein-kininogen system with the severity of the burn and the complement factors BSA BUS c4 c3c 0.27n 0.23* - 0.26 - 0.23 Prokallikrein 0.49* - 0.38X 0.56” - 0.31% Total kininogens 0.46” 0.52% - 0.38X - 0.30% LM W kininogen 0.42* 0.48* - 0.25 - 0.25 HM W kininogen (WI significant at the level p .=z0.05 Correlations at admission
542
KININ SYSTEM IN BURN
Vol.
41,
No. 4
DISCUSSION -----~ Several studies performed in animals (3-6) have shown that proteolytic enzymes and kinins are released in burned tissues. Our results obtained in man agree with these previous findings. We have shown that plasma prokallikrein and kininogens are significantly decreased after an extensive burn of the skin and remain low during the acute inflammatory response. A decreased prokallikrein activity suggests that this enzyme is activated by the injury, releasing bradykinin from HM W kininogen. The plasma concentration of HMW kininogen however is modified in the same way as total proteins. Further, values of LMW kininogen are even more decreased. This apparent discrepancy, also observed in acute pancreatitis (121, can be explained by the nature of the methods used to quantify both kininogens. We recently showed that “in vitro” the heavy chain used to quantify total and LMW kininogen was more sensitive to proteolysis than the histidin rich light chain specific of HMW kininogen (13). This observation is related to the immunogenicity of both kinds of kininogens and agrees with the results of Scott et al. (141, who observed that HMW kininogen can release bradykinin without loosing the procoagulant activity of its light chain. We also noted that the decrease in LMW kininogen was directly proportional to the severity of the burn, indicating that this kind of kininogen might be cleaved by tissue proteases released after the injury. Our findings suggest that kinins liberated early after skin burn participate in the inflammatory response. other biochemical mediators are also involved, e.g., the complement However, and C4 which are simultaneously activated in burned patients as zt%? inc3fhis stu dy. This confirms the results of Green (15) who described some differences between the exsudates induced by bradykinin or by thermal injury and our previous observation that heating induces oedema in the paw of normal rats as well as in brown Norway rats in which kinin formation does not occur (16). REFERENCES -------1.
ADAM, A., ALBERT, A., CALAY, G., CLOSSET, J., DAMAS FRANCHIMONT, P. Human kininogens of low- and high- molecular Quantification by radioirnmunoassay and determination of reference Clin. Chem. 31,- 423-426, 1985. _
2.
ADAM, A., AZZOUZI, M., BOULANGER, J., ERS, P., ALBERT, A., DAMAS. J. and FAYMONVILLE. M-E. Obtimized determination of olasma prokallikrein on a Hitachi 705’ autoanaliser. Clin. Chem. Clin. Biochem. ---------~---&3, 203-207, 1985.
3.
ROCHA e mechanism Med. Exe, --
4.
GARCIA LEME, J., HAMAMURA anti-proteases and hexadimethrine bradykinin-like substance during Pharmacol. 40, 294-309, 1970. --
5.
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