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The kallikrein-kinin system in the brown norway rat
THROMBOSIS RESEARCH 31; 653-656, 1983 0049-3848/83 $3.00 + .OO Printed in the USA. Pergamon Press Ltd. METHODS:
MINI-REPORT
THE KALLIKREIN-KININ SYSTEM IN THE BROWN NORWAY RAT 1 2 Eric Marks, M.D. , Barbara Alving, M.D. , and John J. Pisano, Ph.D.3 1 Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814 n
'Department of Hematology, Walter Reed Army Institute of Research, Washington, D.C. 20012 3 National Institutes of Health, National Heart, Lung and Blood Institute, Laboratory of Chemistry, Section on Physiological Chemistry, Building 10, Room 7N260, Bethesda, Maryland 20205 Received 18.3.83; Accepted in revised form 27.5.83 by Editor O.D. Ratnoff
The kallikrein-kinin system has been implicated in a variety of pathophysiological reactions. Kinins produce pain, increase vascular permeability, relax vascular smooth muscle and lower blood pressure, contract isolated smooth muscle preparations, and stimulate prostaglandin synthesis in cells and target tissues. Furthermore, plasma kallikrein and high molecular weight (HMW) kininogen greatly enhance in vitro contact activation of coagulation. Studies to establish a definitive role for kinins, kallikrein and kininogens have been impeded by the lack of specific blocking agents and suitable animal models. It now appears, however, that an excellent animal model for HMW kininogen and plasma kallikrein deficiency has been found. Damas and Adam reported that the Brown Norway rat (Rattus norvegicus) (BN/Mai Pfd f) is severely deficient in plasma kallikrein and partially deficient in kininogen (1). The congenital deficiency in man characterized by a severe deficiency of HMW kininogen (2) and partial deficiency of LMW kininogen and plasma kallikrein (3) has been named Fitzgerald trait. Using rats obtained from the same source as Damas and Adam, (Katholieke Universitiet of Leuven, Haverlee, Belgium), Oh-ishi et al confirmed the results of Damas and Adam and showed that the rats from Haverlee have only 25-30% the normal level of prekallikrein and that they may be totally deficient in HMW kininogen (4). On the other hand, BN rats from Kitasato, Japan were normal. Because of the potentially great value of the report of Damas and Adam, we have examined Brown Norway rats from MA Laboratory Animals of Harlan SpragueDawley (formerly Microbiological Associates), Walkersville, Maryland. They represent the sole American source and originator of the parent strain, BN/Mai f (5), now classified as BN/NHsd. Sprague-Dawley (SD) rats (Taconic Farms) served as controls. To measure plasma prekallikrein and kininogen we employed both an in vitro enzymatic assay and in vivo assay based on the fall in blood pressure following activation of the rat's prekallikrein. Two activators of
Key Words:
Brown Norway rats, kallikrein, kininogen 653
KALLIKREIN-KININ/BROWN NORWAY RAT
654
Vo1.31, No.4
prekallikrein were employed, dextran sulfate (Pharmacia, MW 500,000) and the Q, 30,000 dalton fragment of Hageman factor (HF or B-XII ) prepared in this laboratory (6). Dextran sulfate is an indirecf activate? which initiates a complex reaction in plasma involving Hageman factor, prekallikrein and high molecular weight kininogen, whereas HFf is a potent direct activator of prekallikrein. When administered intravenously to normal rats, HFf causes the rapid generation of bradykinin and hypotension (7). BN and SD rats showed the same fall in blood pressure when treated with either dextran sulfate or HFf (Table 1). The rats also responded with a hypotensive episode of similar magnitude and duration when given a second injection of dextran sulfate indicating comparable levels of kininogen. In vitro assays of plasma kallikrein and Hageman factor also revealed no difference between BN and SD rats (Table 2).
TABLE 1 Lowering of Blood Pressure by Intravenous Injectiona of Dextran Sulfate or Hageman Factor Fragments into Brown Norway (BN) and Sprague-Dawley (SD) Rats
Strain(n)b
Dextran Sulfate Blood PressureC Injection t post injection decrease pre injection torr torr
SD(4) SD(4) BN(4) BN(4)
1 2 1 2
SD(5) BN(5)
1 1
87t9 104+14 92?7 9726
44+3 65230 3922 3924
DurationC seconds
43 39 53 58
311t111 88225 262+77 118257
36 33
248281 205f97
Hageman Factor Fragments 82+8 78211
46+9 4527
a Animals were anesthetized with sodium pentobarbital, 50 mgikg. The left femoral artery and vein were cannulated with PE-50 tubing and the arterial catheter was advanced to the level of the aortic bifurcation. All animals had stable blood pressures (measured with a P23D6 Stratham transducer + Grass 791)recorder) which were unchanged by painful stimuli of the hind paw. Animals were tested 30 min after surgery. Those that received dextran sulfate were given a second injection 10 min later (i.e. after the blood pressure had stabilized) to indicate the degree of kininogen depletion. Doses employed were 0.28 mg dextran sulfate and 77 ng Hageman factor fragments. b
n is the number of animals tested.
' Data are presented as mean + standard deviation. Differences between the strains were insignificant (p>O.O5, analysis of variance).
Vo1.31, No.4
KALLIKREIN-KININ/BROWN NORWAY RAT
655
TABLE 2 Comparison of Plasma Levels of Prekallikrein and Hageman Factor in Sprague-Dawley and Brown Norway Rats
Strain (n)b
Prekallikreina Dextran Sulfate HFf units/ml
Sprague-Dawley (8) Brown Norway (7)
0.76kO.22' 0.6OkO.16
0.61kO.19 0.59+0.12
Hageman Factor % of Normal Human Pool 204+57 203+50
aplasma prekallikrein was assayed after activation with dextran sulfate (8) or purified HFf (9) with the substrate D-prolyl-Lphenylalanyl-L-arginyl-p-nltranilide (S-2302 of Kabi). Dextran sulfate is an indirect activator and HFf a direct activator of prekalllkrein. Factor XII was measured by correction of the partial thromboplastln time of human plasma deficient in Factor XII, and values were compared to the level in normal human blood (10). b n is the number of animals tested. 'Data are presented as mean + standard deviation. Differences between the strains were insignificant (p>O.OS, Student t-test).
We conclude that BN rats obtained from the original parent source, MA Laboratory Animals of Harlan Sprague-Dawley, have normal levels of plasma prekallikrein and kininogen, unlike those at the Katholieke Universitiet of Leuven (Haverlee). Increased availability of these rats and their distribution should greatly aid research on the pathophysiologic significance of the kallikrein-kinin system.
REFERENCES 1.
DAMAS, J. and ADAM, A. Congenital deficiency in plasma kallikrein and kininogens in the brown Norway rat. Experientia,a, 586-587, 1980.
2.
SAITO, H., RATNOFF, O.D., WALDMANN, R., and ABRAHAM, J.P. Fitzgerald trait. Deficiency of a hitherto unrecognized agent, Fitzgerald factor, participating in surface-mediated reactions of clotting, fibrinolysis, generation of kinins, and the property of diluted plasma enhancing vascular permeability (PF/Dil). .J Clin Invest.55_, 1082-1089, 1975.
3.
OH-ISHI, S., UENO, A., UCHIDA, Y., KATORI, M., HAYASHI, H., KOYA, H., KITAJIMA, K. and KIMURA, I. Abnormalities in the contact activation through factor XII in Fujiwara trait: A deficiency in both high and low molecular weight klninogens with low level of prekallikrein. Tohoku J EXP Med.u, 67-80, 1981. -.-
656
KALLIKREIN-KININ/BROWN NORWAY RAT
Vo1.31, No.4
4.
OH-ISHI, S., SATOH, K., HAYASHI, I., YAMAZAKI, K. and NAKANO, T. Differences in prekallikrein and high molecular weight kininogen levels in two strains of brown Norway rat (Kitasato Strain and Katholiek Strain). Thrombosis Res. 28 143-147, 1982. .-I
5.
Animals for Research: A Directory of Sources. 10th Edition. Institute of Laboratory Animal Resources, National Research Council, National Academy of Sciences, 1979, p 37.
6.
HOJIMA, Y., TANKERSLEY, D.L., MILLER-ANDERSON, M., PIERCE, J.V. and PISANO, J.J. Enzymatic properties of human Hageman factor fragment with plasma prekallikrein and synthetic substrates. Thrombosis Res.18, 417-430, 1980.
7.
MARKS, E., HOJIMA, Y., FRECH, M.E., KEISER, H. and PISANO, J.J. An inhibitor from corn blocks the hypotensive action of plasma protein fraction and active Hageman factor. Thrombosis Res.23, 97-102, 1980.
8.
KLUFT, C. Determination of prekallikrein in human plasma: optimal conditions for activating prekallikretn. J Lab and Clin Med.9l, 83-95, 1978.
9.
ALVING, B.M., TANKERSLEY, D.L. and MASON, B.L. Plasma prekallikrein: quantitative determination by direct activation with Hageman Factor fragment (@XII,). J Lab and Clin Med l&, 226-241, 1983.
10.
GIDDINGS, J.C. Hereditary coagulation disorders: laboratory techniques In: Blood Coagulation and Haemostasis. A Practical Guide. J.M. Thomson _-._... _I.__-______ (Ed) New York: Churchill Livingstone, 1980, pp 144-146.
The opinions and assertions contained herein are those of the author and are not be construed as representing those of the Uniformed Services University of the Health Sciences or the Department of Defense.
MINI-REPORT
THE KALLIKREIN-KININ SYSTEM IN THE BROWN NORWAY RAT 1 2 Eric Marks, M.D. , Barbara Alving, M.D. , and John J. Pisano, Ph.D.3 1 Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814 n
'Department of Hematology, Walter Reed Army Institute of Research, Washington, D.C. 20012 3 National Institutes of Health, National Heart, Lung and Blood Institute, Laboratory of Chemistry, Section on Physiological Chemistry, Building 10, Room 7N260, Bethesda, Maryland 20205 Received 18.3.83; Accepted in revised form 27.5.83 by Editor O.D. Ratnoff
The kallikrein-kinin system has been implicated in a variety of pathophysiological reactions. Kinins produce pain, increase vascular permeability, relax vascular smooth muscle and lower blood pressure, contract isolated smooth muscle preparations, and stimulate prostaglandin synthesis in cells and target tissues. Furthermore, plasma kallikrein and high molecular weight (HMW) kininogen greatly enhance in vitro contact activation of coagulation. Studies to establish a definitive role for kinins, kallikrein and kininogens have been impeded by the lack of specific blocking agents and suitable animal models. It now appears, however, that an excellent animal model for HMW kininogen and plasma kallikrein deficiency has been found. Damas and Adam reported that the Brown Norway rat (Rattus norvegicus) (BN/Mai Pfd f) is severely deficient in plasma kallikrein and partially deficient in kininogen (1). The congenital deficiency in man characterized by a severe deficiency of HMW kininogen (2) and partial deficiency of LMW kininogen and plasma kallikrein (3) has been named Fitzgerald trait. Using rats obtained from the same source as Damas and Adam, (Katholieke Universitiet of Leuven, Haverlee, Belgium), Oh-ishi et al confirmed the results of Damas and Adam and showed that the rats from Haverlee have only 25-30% the normal level of prekallikrein and that they may be totally deficient in HMW kininogen (4). On the other hand, BN rats from Kitasato, Japan were normal. Because of the potentially great value of the report of Damas and Adam, we have examined Brown Norway rats from MA Laboratory Animals of Harlan SpragueDawley (formerly Microbiological Associates), Walkersville, Maryland. They represent the sole American source and originator of the parent strain, BN/Mai f (5), now classified as BN/NHsd. Sprague-Dawley (SD) rats (Taconic Farms) served as controls. To measure plasma prekallikrein and kininogen we employed both an in vitro enzymatic assay and in vivo assay based on the fall in blood pressure following activation of the rat's prekallikrein. Two activators of
Key Words:
Brown Norway rats, kallikrein, kininogen 653
KALLIKREIN-KININ/BROWN NORWAY RAT
654
Vo1.31, No.4
prekallikrein were employed, dextran sulfate (Pharmacia, MW 500,000) and the Q, 30,000 dalton fragment of Hageman factor (HF or B-XII ) prepared in this laboratory (6). Dextran sulfate is an indirecf activate? which initiates a complex reaction in plasma involving Hageman factor, prekallikrein and high molecular weight kininogen, whereas HFf is a potent direct activator of prekallikrein. When administered intravenously to normal rats, HFf causes the rapid generation of bradykinin and hypotension (7). BN and SD rats showed the same fall in blood pressure when treated with either dextran sulfate or HFf (Table 1). The rats also responded with a hypotensive episode of similar magnitude and duration when given a second injection of dextran sulfate indicating comparable levels of kininogen. In vitro assays of plasma kallikrein and Hageman factor also revealed no difference between BN and SD rats (Table 2).
TABLE 1 Lowering of Blood Pressure by Intravenous Injectiona of Dextran Sulfate or Hageman Factor Fragments into Brown Norway (BN) and Sprague-Dawley (SD) Rats
Strain(n)b
Dextran Sulfate Blood PressureC Injection t post injection decrease pre injection torr torr
SD(4) SD(4) BN(4) BN(4)
1 2 1 2
SD(5) BN(5)
1 1
87t9 104+14 92?7 9726
44+3 65230 3922 3924
DurationC seconds
43 39 53 58
311t111 88225 262+77 118257
36 33
248281 205f97
Hageman Factor Fragments 82+8 78211
46+9 4527
a Animals were anesthetized with sodium pentobarbital, 50 mgikg. The left femoral artery and vein were cannulated with PE-50 tubing and the arterial catheter was advanced to the level of the aortic bifurcation. All animals had stable blood pressures (measured with a P23D6 Stratham transducer + Grass 791)recorder) which were unchanged by painful stimuli of the hind paw. Animals were tested 30 min after surgery. Those that received dextran sulfate were given a second injection 10 min later (i.e. after the blood pressure had stabilized) to indicate the degree of kininogen depletion. Doses employed were 0.28 mg dextran sulfate and 77 ng Hageman factor fragments. b
n is the number of animals tested.
' Data are presented as mean + standard deviation. Differences between the strains were insignificant (p>O.O5, analysis of variance).
Vo1.31, No.4
KALLIKREIN-KININ/BROWN NORWAY RAT
655
TABLE 2 Comparison of Plasma Levels of Prekallikrein and Hageman Factor in Sprague-Dawley and Brown Norway Rats
Strain (n)b
Prekallikreina Dextran Sulfate HFf units/ml
Sprague-Dawley (8) Brown Norway (7)
0.76kO.22' 0.6OkO.16
0.61kO.19 0.59+0.12
Hageman Factor % of Normal Human Pool 204+57 203+50
aplasma prekallikrein was assayed after activation with dextran sulfate (8) or purified HFf (9) with the substrate D-prolyl-Lphenylalanyl-L-arginyl-p-nltranilide (S-2302 of Kabi). Dextran sulfate is an indirect activator and HFf a direct activator of prekalllkrein. Factor XII was measured by correction of the partial thromboplastln time of human plasma deficient in Factor XII, and values were compared to the level in normal human blood (10). b n is the number of animals tested. 'Data are presented as mean + standard deviation. Differences between the strains were insignificant (p>O.OS, Student t-test).
We conclude that BN rats obtained from the original parent source, MA Laboratory Animals of Harlan Sprague-Dawley, have normal levels of plasma prekallikrein and kininogen, unlike those at the Katholieke Universitiet of Leuven (Haverlee). Increased availability of these rats and their distribution should greatly aid research on the pathophysiologic significance of the kallikrein-kinin system.
REFERENCES 1.
DAMAS, J. and ADAM, A. Congenital deficiency in plasma kallikrein and kininogens in the brown Norway rat. Experientia,a, 586-587, 1980.
2.
SAITO, H., RATNOFF, O.D., WALDMANN, R., and ABRAHAM, J.P. Fitzgerald trait. Deficiency of a hitherto unrecognized agent, Fitzgerald factor, participating in surface-mediated reactions of clotting, fibrinolysis, generation of kinins, and the property of diluted plasma enhancing vascular permeability (PF/Dil). .J Clin Invest.55_, 1082-1089, 1975.
3.
OH-ISHI, S., UENO, A., UCHIDA, Y., KATORI, M., HAYASHI, H., KOYA, H., KITAJIMA, K. and KIMURA, I. Abnormalities in the contact activation through factor XII in Fujiwara trait: A deficiency in both high and low molecular weight klninogens with low level of prekallikrein. Tohoku J EXP Med.u, 67-80, 1981. -.-
656
KALLIKREIN-KININ/BROWN NORWAY RAT
Vo1.31, No.4
4.
OH-ISHI, S., SATOH, K., HAYASHI, I., YAMAZAKI, K. and NAKANO, T. Differences in prekallikrein and high molecular weight kininogen levels in two strains of brown Norway rat (Kitasato Strain and Katholiek Strain). Thrombosis Res. 28 143-147, 1982. .-I
5.
Animals for Research: A Directory of Sources. 10th Edition. Institute of Laboratory Animal Resources, National Research Council, National Academy of Sciences, 1979, p 37.
6.
HOJIMA, Y., TANKERSLEY, D.L., MILLER-ANDERSON, M., PIERCE, J.V. and PISANO, J.J. Enzymatic properties of human Hageman factor fragment with plasma prekallikrein and synthetic substrates. Thrombosis Res.18, 417-430, 1980.
7.
MARKS, E., HOJIMA, Y., FRECH, M.E., KEISER, H. and PISANO, J.J. An inhibitor from corn blocks the hypotensive action of plasma protein fraction and active Hageman factor. Thrombosis Res.23, 97-102, 1980.
8.
KLUFT, C. Determination of prekallikrein in human plasma: optimal conditions for activating prekallikretn. J Lab and Clin Med.9l, 83-95, 1978.
9.
ALVING, B.M., TANKERSLEY, D.L. and MASON, B.L. Plasma prekallikrein: quantitative determination by direct activation with Hageman Factor fragment (@XII,). J Lab and Clin Med l&, 226-241, 1983.
10.
GIDDINGS, J.C. Hereditary coagulation disorders: laboratory techniques In: Blood Coagulation and Haemostasis. A Practical Guide. J.M. Thomson _-._... _I.__-______ (Ed) New York: Churchill Livingstone, 1980, pp 144-146.
The opinions and assertions contained herein are those of the author and are not be construed as representing those of the Uniformed Services University of the Health Sciences or the Department of Defense.