The Effects of Bile Salts on Some Coagulation Components and Related Enzymes^[*]

Summary

Certain purified bile salts, individually or in a mixture, profoundly affect - either inhibiting or enhancing - the esterolytic activities of thrombin, trypsin, and plasmin, the clotting activity of thrombin, and caseinolysis by trypsin. They also promote SK-induced fibrinolysis and impair FI clottability. These effects are directly related to bile salt concentration but not to the critical micellar concentration.

A very unusual effect was observed with deoxycholate and FI : besides inhibiting FI clottability, the salt induces spontaneous gelation. In addition, it binds strongly to the protein, as has been already reported for another plasma protein, albumin, and to a lesser degree, to alpha- and beta-globulins.

Noteworthy is the fact that activation of pancreatic juice trypsinogen by thrombin also was increased by prior thrombin exposure to the salts. On the other hand, thrombin-induced platelet aggregation was slightly inhibited by the bile salt mixture, which, when added to PRP moderately inhibited the ADP-induced aggregation. No effect was observed on the conversion of F II in plasma via the thromboplastic mechanism when deoxycholate, or cholate, or glycocholate was added to the system.

It is postulated that the bile salt mixture enhances SK-induced fibrinolysis by direct action, either on SK, or on the SK-activator complex, attributable to the detergent properties of the salts.

The physiologic and pathologic implications of our results with respect to hemostasis and pancreatitis are discussed.

^* This investigation was supported by U.S. Public Health Service Grants HE-11447 and HE-09011. A preliminary description of this work was presented at the Meetings of the Federation of American Soc. E xperimen. Biology, Atlantic City, N .J., 1968.

• References

• 1 Hofmann A. F. The function of bile salts in fat absorption. Biochem. J 89: 57 1963;
• 2 Garagnani A, Facchini G. Influenza della bile e dei sali biliari sul l’emocoagulazione in vitro con particolare riguardo all* attivita antitrombinica. Arch. Patol. Clin. med 33: 203 1956;
• 3 Curragh E. F, Elmore D. T. Kinetics and mechanism of catalysis by proteolytic enzymes. 2. Kinetic studies of thrombin-catalysed reactions and their modification by bile salts and other detergents. Biochem. J 93: 163 1964;
• 4 Engel A. M, Pechet L, Alexander B. Activation of trypsinogen and plasminogen by thrombin preparations. Biochem 05: 1543 1966;
• 5 Engel A. M, Alexander B. Activation of chymotrypsinogen-A by thrombin preparations. Biochem 05: 3590 1966;
• 6 Alexander B. Recent studies on plasma colloid substitutes: Their effects on coagulation and hemostasis. In: Body Fluid Replacement in the Surgical Patient. Conference Chairmen, Charles L.Fox, Jr. and Gabriel G.Nahas. 157 Grune & Stratton: New York; 1970
• 7 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Arkiv for Kemi 10: 415 1956;
• 8 Seegers W, McGlaughry R, Fahey J. Some properties of purified prothrombin and its activation with sodium citrate. Blood 05: 421 1950;
• 9 Sjövall J. On the concentration of bile acids in the human intestine during absorption. Acta physiol, scand 46: 339 1959;
• 10 Norman P. Studies of the plasmin system. I. Measurement of human and animal plasminogen. Measurement of an activator in human serum. J. Exp. Med 106: 423 1957;
• 11 Kattlove H. E, Alexander B. The effect of cold on platelets. I. Cold-induced platelet aggregation. Blood 38: 39 1971;
• 12 Division of Biological Control, National Institutes of Health. Second Revision. September 1946
• 13 Ware A. G, Seegers W. H. Two stage procedure for the quantitative determination of prothrombin concentration. Amer. J. Clin. Path 19: 471 1949;
• 14 Hummel B. C. W. A modified spectrophotometric determination of chymotrypsin, trypsin and thrombin. Can. J. Biochem. Physiol 37: 1393 1959;
• 15 Hofmann A. F. Thin layer chromatography of free and conjugated bile acids on silicic acid. J. Lipid Research 03: 127 1962;
• 16 Lowry O. H, Rosebrough N. J, Farr A. L, Randall R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem 193: 265 1951;
• 17 Born G. V. R. Quantitative investigations into aggregation of blood platelets. J. Physiol 162: 67 1962;
• 18 Olesen E. S. Activation of fibrinolysis in guinea pig serum by bile acids. Thrombos. Diathes. haemorrh. (Stuttg) 04: 473 1960;
• 19 Dam H, Kruse J, Krog Jensen M, Kallenhange H. E. Studies on human bile. II. Influence of two different fats on the composition of human bile. Scand. J. clin. Lab. Invest 19: 367 1967;
• 20 Wainio W. W. Properties of heart muscle cytochromes prepared with deoxycholate. Trans. N. Y. Acad. Sci 22: 426 1960;
• 21 Barthels M, Seegers W. H. Substitution of lipids with bile salts in the formation of thrombin. Thrombos. Diathes, haemorrh. (Stuttg) 12: 13 1969;
• 22 Rudman D, Kendall F. E. Bile acid content of human serum. II. The binding of cholanic acids by human plasma proteins. J. clin. Invest 36: 538 1957;
• 23 Astrup T, Allcjaersig N. Quaternary detergents and the activity of trypsin and plasmin. Nature 168: 565 1951;
• 24 Richard M. N, Sanders B. E. Effect of lysine and wetting agents on activated plasminogen solutions. Canad. J. Biochem 41: 211 1963;
• 25 Kwaan H. G, Brahman P, Astrup T. Enhancement of fibrinolysis and thrombolysis by polysorbate 80 (Tween 80). Experientia (Basel) 23: 1 1967;
• 26 Lach G. H, Yousuf Ali S. Tissue activator of plasminogen. Nature 201: 1030 1964;
• 27 Cole E. R. Hydrolysis of L-histidine methylester. I. The action of thrombin Thrombos. Diathes. haemorrh. (Stuttg) 19: 321 1968;
• 28 Jirgenson B. Effect of detergents in the conformation of proteins. II. Optical rotatory dispersion by using ultraviolet light Arch. Biochem. Biophys 96: 321 1962;
• 29 Lewis B, Tabaqchali S, Panveliwalla D, Wootton I. D. P. Serum-bile acids in the stagnant-loop syndrome. Lancet 01: 219 1969;
• 30 Carey J. B. The serum trihydroxy-dihydroxy bile acid ratio in liver and biliary tract disease. J. clin. Invest 37: 1494 1958;