nounced changes were observed in the seromucoid, haptoglobin, and ceruloplasmin fractions. These fractions started to rise in the first 4 days after the fracture, ...
Dynamic Changes of Different Serum Glycoproteins after Bone Fractures Mala Herzberg,
Z. Oberman,
S. L. Weissman, and H. Z. Herold
The physiologic response to a simple injury was studied in a group of 24 adults suffering from uncomplicated fractures of the tibia and fibula. The dynamic changes of different protein and glycoprotein fractions were estimated. At Days 1, 4, 10, and 30 after the fracture, there was no significant alteration in the total protein levels,
but the albumin decreased (18%) and and z2-globulins increased. The most pronounced changes were observed in the seromucoid, haptoglobin, and ceruloplasmin fractions. These fractions started to rise in the first 4 days after the fracture, but only the seromucoid and ceruloplasmin remained elevated after a month. The results suggesta rapid turnover of thoseglycoprotein fractions; they also show that even a
local injury,
like an uncomplicated fracture of the tibia and fibula, is sufficient to cause
significant elevation of most of the glycoprotein fractions. The possible mechanism
is discussed.
IN
OUR PREVIOUS PAPERS
elevation of a-globulins As the glycoproteins decided to investigate proteins
in or(ler
the influence healthy
adult
(1-3) it is shown that there was a significant in patients who had sustained bone fractures. consist of a group of conjugated proteins, we the behavior of many subfractions of glyco-
to get a clearer
of trauma. subjects
picture
of their
dynamic
changes
under
this purpose we chose a group of otherwise of whom was admitted to the orthopedic a simple and uncomplicated fracture of the tibia For each
ward suffering from and fibula. This group served as a model system to study physiologic reactions arising after trauma in a limited region of the body without concomitant inflammation. From the total glycoproteins we chose the seromucoid fractions as a representative indicator of 1-globulins; for the glycoproteins moving electrophoretically with 0-globulins, we determined haptogiobni and ceruloplasniin. As a representative of hexose-rich components of $-globulins, we chose fibrinogen. We also measured the red cell sedimentation rate (ESR) since it is a function of the changes in plasma proteins. From
the
Department
Ichilov Municipal Hospital, Received for publication
of
Biochemistry and the Department of Orthopedic Tel-Aviv University Medical School, Israel. Mar. 20, 1967; accepted for publication May 5, 1967. Clinical
1065
Surgery,
.0
t.-.
.0cc
cc.0
-4
-4 -
-
CCCCCOciC
C
C
C
C
C
©
-
-
C
-
.0
-
C
‘
4
C C
tc
-
cc C -© c-i dCClc-iC
-H II A -H II V 41 II V -H II A -ft II A -H II A -H II A 41 II V -H II A -H II V -H II A -H II V O
0.
“
C
0
c-i c-i ‘0
C
C
C
C
C
.0
-H IA-HIP
-,
cc C
C
C
cc
V-Hit
-
c-I
-
t..
C
C
-
C
.0
-
40
C
C
cc ©
v-HIP
V-HIP
-
4
C cc
C
-
.0
C
©
-
V-H
-
c-i cc C
t-
-
c-i ‘0
C
V-HIP
VA-HO
-
0.
,
,
-
0.
cc cc
t=
cc so
C.
‘©-
C
-
cc
C
-
V
-H
V-HIP
V-HO
cO’0
-
‘0
cc © c-i ©
II V
co
C
41 II
V 0.
©
I
©
-4
‘0
-
50
-
“
t..
‘
-
cc
-
C
-
0
cc ‘0
“4
.‘
t-.
‘0
Cc
‘
.-
cc
-
‘O C
-
‘0
-
-H
II A
-H
-H
II V
II V
-H
C
II V
-H
-H
II V
II A
-H
II V
-H
II V
-H
It V
-H
cc’0
-
CCCCe-CCCCccCCc-iCC-4C-’0Cc-i’0C.00C-4C’i
II V
-
t’-
-H
IC
CO
-H
II A
0.-’
cc
cc
C
‘0
‘0
C
-
c-i
-
©
-
.0
CO -
CO
ci
‘0
CO ‘0
‘0
‘0
cc
-
ci
t-.
-
‘
cc
-4
C
‘
a
-‘
-H
C
C
P
C
C
‘
-H
II A -H II V
C
II V
C
-H
40
C
II V
C
-
C
-H
C
-
-H
II A
C
II A
-
cc
41
C
II V
-
c-i
-H
C
II V
cc
#{149}CiC
-H
I.,-
C
C
II A
0.
‘0
‘0
.O0©#{149}OC’4
o
C
II V
c-c
C
C
-4
C
4
C
C
-H
II
1’
-H
-C -
-
-
C
II A
C
-
-H
c-i
C
II V 0,
0.
c-c
c-i 1
‘0 c-
C
#{149}0
-
cc
-
C
cc
c-i
-
-
11
1
-H
c-i
c-i ci
.0
-
C
C
-H
41
-
8
.S. aEE
-
.0
o
.E
n -
.0
-
.08 .9bc
so
c-i
t-
-
-
“4
-H
-H
-H
C “4
CO
C
‘0
cc
C C
C
-H
-4
-
ci cc
il
.
2.g
Tdr-..8e
g
‘ 5-
h .
.s
E E
8b -
0
c-i
cc
.E
*a’
E -
C
-
-H
ci
-
-
-
-
C,
-
cc c-c
ci
-ft
.
8
‘
c--i
I-
-
o
an
ci cc
0
CO
C
1
‘0
C
r12
Vol. 13, No. 12, 1967
SERUM GLYCOPROTEINS
1067
Materials and Methods A total of 24 patients (16 men and 8 women aged from 20-60 years) who had sustained an uncomplicated fracture of the tibia and fibula were repeatedly examined for a period of 1 month beginning with the day of admission to the hospital. Blood specimens were collected after overnight fasting on Days 1 or 2, 4 or 5, 10, and approximately 1 month after the fracture. We measured: (1) total protein by the method of Weichselbaum (4); (2) electrophoretic pattern of erum proteins according to Bloch et at. (5); (3) glycoprotein (hexose-bound protein) by the method of Shetlar et at. (6); (4) seromucoid according to the method of Winzler et at. (7); (5) haptoglobin by the method of Juva and Laurent (8); (6) ceruloplasmin according to Ravin (9); (7) fibrinogen using a modification of the method of Quick (10); and (8) the Westergren red cell sedimentation rate.
Results Table 1 presents the results of the investigation with statistical evaluation of significance according to the t test. As shown in Table 1 the total protein and y-globulin fraction did not change significantly. Relatively large changes, like those reported in our previous publications, occurred in the and 2-globulin fractions, which were elevated. There was a concomitant drop in the albumin level. The red cell sedimentation rate was elevated throughout the period of investigation. The behavior of various glycoproteins is particularly interesting. The total glycoprotein measured as hexose-bound protein was elevated by 18% at 4-S (lays after tile fracture occurred (Table 2) and had returned to normal within 1 month. The haptoglohin, which is the major component of the 0-g1obulin-bound hexoses, had the greatest elevation from Days 4 to 10 after fracture, and then returned to normal after 1 month. By contrast, ceruloplasmin, which also contributes to the glycoproteins moving electrophoretically in the z2-globulin fraction, rose 20% on Days 4-5 after the fractures and remained elevated even a month later (17.5%). The seromucoid-which is the fraction of the serum richest in carbohydrate-was highly elevated the day following the fracture (16%) and remained elevated to the end of the investigation period (36.5%). Fibrinogen, moving with $-globulin fraction, was found to be elevated on Day 10 following the fracture (26%) and returned to normal levels after 1 month. Concerning the glycoprotein fractions, the dynamic changes are as -
1068
HERZBERG
Table 2.
DEVIATIONS
OF PROTEIN NORMAL
FT AL.
ClinkalChemistry
AND GLYCOPROTEIN MEAN
FRACTIONS
0
Parameter.
‘rota! protein Albumin a,-globulin ai-globulin fl-globulin
-y-globulin Hexose-bound Seromucoid Haptoglobin eruIoplasmin Fibrinogen
protein
D4zy.
AS
%
Os’
VAlUES
1‘
D41y0 4-3
1-2
32 -16.7 +385 +29.5 + 7.4 + 7.9 +11.0
+16.0 69 0 9io
10
03
1.7 -18.8 +59.0 +36.5 +17.4 + 55 +18.5 +42.0 +57.0 +20.0
-12.8 +32.0 +25.5 +19.4 + 7.1 +10.5 +52.0 +64.5 +27.5
+
+26-0
-
+
Day -
Day
30
+
L4
-
3.3
+13.5 + 4.9 + 4.6 + 7.9 + 38 +36.5
+15.2 +17.5 -
1.3
follows:
Total glycoprotein and seromucoid started to rise the day the fracture, but only after 4 days did we observe a significant elevation of haptoglobin and ceruloplasmin. The above findings suggest a fast turnover of most of the glycoprotein fractions in human serum which is in agreement with the work of Weisman et al. (ii), Winzler (12), and Nyman (13). following
Discussion The above results show that even local trauma caused by uncomplicated fracture of the tibia and fibula produces drastic changes in distinct protein and glycoprotein fractions, despite the fact that the total protein values remain in the normal range. The results also show a good correlation between the elevation of and $-globulins and their consti tuent carbohydrate-rich components. Studies made by Macbeth et at. (14) have showii that the liver is the major, if not the sole .site of serum glycoprotein biosynthesis, and that the rate of biosynthesis and degradation of the seromucoid fraction is more rapid than that of the total serum glycoprotein. Our results show that the seromucoid and ceruloplasmin remain elevated even 1 month after the fracture. It is therefore possible to postulate that the healing process of fractures is accompanied by an acceleration in the formation of haptoglobin, seromucoid, and ceruloplasmin, of which the last two remain elevated a relatively long time. It is known that in most inflammatory diseases there is an elevation of and y-globulin fractions. Tn our group of uncomplicated fractures, we observed an increase only in -globuliris, while the y-globulins remained at an almost normal level. This may denote a lack of inflammation in the simple, uncomplicated bone injury. -
-
V01. 13, N0. I,
1967
SERUM GLYCOPROTEINS
1069
The elevation of glycoproteins in the serum seen after fractures cannot be explained by the suggestion of M#{228}rki and Wuhrmann (is) who postulated that the loss of albumin in acute inflammatory disease is compensated by a-globulins and fibrinogen so as to maiiitain circulatory function through a constant blood volume. Our results show that tile maximal decrease in albumin (Table 2) was 18%. To replace the amount of albumin while maintaining the total protein in the normal range, other fractions must be increased. We found significant increases only in and c2-globuliii fractions. This can be interpreted as a very rapid turnover of these fractions and suggests that the fractions rich in glycoproteins have an additional function in recovery after traumatic injury. More pronounced are the dynamic changes in the seromucoid and haptoglobin fractions, which increased to a maximum of 52% and 64.5%, -
respectively.
it is known
bind easily to hemoglobin and that they disappear from the circulation in cases of acute hemolysis. It is clear, therefore, that in uncomplicated fractures there is no sign of hemolysis. The elevation of haptoglobiiis after injury must have another physiologic significance and their role is still obscure. that
tile haptoglobiios
Keyser (16) and Winzler (12) suggest that the elevation of glycoproteins represents a nonspecific response to tissue damage. This response possibly may be mediated by liberation of breakdown products from the necrotic tissue. According to Winzler (12) “these agents act on the liver in such a way as to induce the formation of messenger RNA specific for certain proteins, including the glycoproteins.” The kind of tissue breakdown products that act as stimulators for glvcoprotein synthesis in the liver, the role of glycoprotein in overcoming the disease process, and the determination of whether there is a hormonal control of these glycoproteins are questions that remain for further study.
References 1. Hertzberg, M., Obernian, Z.,Weissman, S. L., and Herold, H. Z, Serum glycoproteins and proteins after fractures. Clin. Cheia. 11, 920 (1965). 2. Weissman, S. L., Herold, H. Z., Hertzberg, M., and Oherman, Z, Variation in blood protein fractions during the healing of fractures. J. Intern. Coil. Surgeons 43, 169 (1965). 3. Weissman, S. L., Herold, H. Z., Hertzberg, M., and Oberman, Z., Variations in blood protein fractions after trauma. Harefuah 68, 53 (1965). 4. Weichselbaum, T. T., An accurate and rapid method for determination of proteins in small amounts of blood serum and plasma. Am. J. C!in. Path. 10, 40 (1946). 5. Bloch, R. J., Durrum, E. L., and Zweig, 0., Acad. Press, New York, 1958, 509 pp. 6. Shetlar, M. R., Foster, J. V., and Everett, M. R., Determination of serum polysaecharides by the tryptophane reaction. Proc. Soc. Exp. Bid. Med. 67, 125 (1948).
1070 7. 8.
HERZBERG FT AL.
Clinical
Chemhtry
Winzler, it. J., Devor, A. W., Mehl, I. W., and Smyth, I. M., Studies on the mucoproteins of human plasma. I. Determination and isolation. J. Clin. Invest. 27, 609 (1948). Juva, K., and Laurent, B., Determination of the haptoglobin-hemoglobin complex in human serum with the aid of gel filtration. Scand. J. Lab. Invest. 15, Suppl. 76, 74
(1963). 9. 10. 11. 12. 13. 14. ]5. 16.
Havin, H. A., An improved coloriinetric enzymatic assay of ceruloplasmin. J. Lab. Clin. Med. 58, 161 (1961). Quick, A. J., The Physiology and Pathology of Haemostasis. Kimpton, London, 1951. Weisman, S., Goldsmith, B., Winzler, R. J., and Lepper, M., Turnover of plasma orosomucoid in man. J. Lab. Clin. Med. 57, 7 (1961). Winzler, 11.J., Metabolism of glycoproteins. Clin. Chein. 11, 339 (1965). Nyman, M., Serum haptoglobin, methodical and clinicalstudies. .Scand. J. Clin. Lab. Invest. 2, Suppl. 39 (1959). Macbeth, R. A. L., Bekesi, J. 0., Sugden, E., and Bice, 5., The metabolism of plasma glyeoproteins.J. Biol. Chein. 240, 10 (1965). Mhrki, H. H., and Wuhrmann, F., Plasmavolumen und Plasmaproteine bei akut-entzundlichen Erkrankungen. Schweiz. Med. Wochachr. 95, 736 (1965). Keyser, J. W., Studies of the blood serum glycoproteins in disease and after injury. J. Oklahoma State Med. Assoc. 53, 769 (1960).
