Some clinical aspects of arylsulphatase activity

CLINICA

CHIMICA

REVIEW

SOME

ACTA

ARTICLE

CLINICAL

ASPECTS

L. hf. JXiX4EOSZY~SKl Biochemistry ( Received

APD J

Depurtment, July

zIst,

Mikotaj

OF ARYLSULPHATASE

ACTIVITY

GNIOT-SZI:LzYCKA Kopernik

University,

Tovufi

(Poland)

1966)

The chemical composition of blood and urine reflects to some degree the physiological status of the organism concerned and hence the great value of blood and urine analysis in clinical diagnostics. The last decade witnessed a tremendous interest in the determination of the activities of several enzymes in blood serum and urine and in the observed relationship between those activities and several diseases 1-4. Today the estimation in blood serum of such enzymes as transaminases, lactic and malic acid dehydrogenases, aldolase, acid and alkaline phosphatases constitutes a great help in the differential diagnosis of a number of diseases. Great hopes are also attached to the discovery of isoenzymes and the organ specificity of their spectrum3+. This paper is concerned with the activity of arylsulphatases in various functional states of the organism. Three arylsulphatases are known: two of them, A and B, are lysosomal enzymes and are soluble in water, the third one, C is insoluble and in the cell appears in the microsomal fraction. The first two have greater affinity for z-hydroxy+nitrophenyl sulphate (NCS) and lower for p-nitrophenyl sulphate (NPS), while third one, C, has greater affinity for the latter substrate (NPS). ARYLSULPHATASE

ACTIVITY

IN BLOOD

AND

URINE

Arylsulphatase can be found in blood serum 0-17as well as in urine6,7.9-Jl.14,1R. Its activity is rather low in those media and depends on the substrate used for the tstimation. Some of the activities found by various authors are summarised in Table I. The highest values for arylsulphatase activity were obtained using e-hydroxy5-nitrophenyl sulphate as substrate. In the presence of other substrates, such as $-nitrophenyl sulphate, $-acetophenyl sulphate or phenyl sulphate, only very low activities were shown. The use of @-nitrophenyl sulphate is further complicated by its nonenzymic hydrolysis caused by an unknown factor 9. The high affinity of the arylsulphatase found in blood and urine to 2-hydioxy5-nitrophenyl sulphate and the low one to the other substrates indicates the presence in those media of the soluble arylsulphatases (type II). Dodgson and Spencer lo applying paper electrophoresis to human urine demonstrated the presence of two soluble arylsulphatases in that fluid. No similar data are available as regards blood serum. According to Ammon and Neye the amount of arylsulphatase excreted in urine Clin.

Chim.

Acta,

15 (1967)

381-386

2.

!A 8

T

w

ARYLSULPHATASE

IN

Wenclewski

rg57

and Lewicki,

1963

5.75 5-8

5 8 --~.

5.5

0.2

NCS

5.6

NC5

Dzialoszyriski, Friihlich and Kontek, rg6G NCS -----“.-~-~.. .__ _______ _-. _

Poczekaj,

Dzialoszyfiski,

NCS

40 (serum) 30 (urine)

8

NPS

Dodgson and Spencer, 195,

7.5

5

I I

I8 20

5.X 6.9

17

XCS

.-2PS

r955

24

2%

24

‘L-20

20

20-24

Boyland, Wallace and Williams,

5.8 6.9

IO

__--_

5

5.8

VARIOUS

INVESTIGATORS

---

---

37”

37D

37”

38

38’

38’

37”

37O

37’>

Temp.

_

-

2.50 (1.8%4.Jh)

-..--.. .__~--

-’ -_-

0.0z1~0.0042 2.30 (1.5-3.0) 2.50 (1.5-4.0)

(o.o-o.rg) (0.0-0.21) (0.0~0.2.3) (2.8-7.8) (1.2-6.5)

(cm-0.82)

-

0.06 (o.o- 0.13) -0.10 (0.03-0.20) 0.18 (o.o7-0.26) 8. IO (4.30-I I .9) (A‘+0 (3.40-10.3) -

o.og 0.14 0.13 4.40 4.70

0.39

(0.0-0.02) (o.o-0.02) (1.2-j.30)

-(o,og-*.g7)

(o.og-- 1.50)

srtlphate.

0. I I (0.0-0.21)

x.67

NCS = r-hydroxy-5-nitrophenyt

BY

Time h

NPS XPS

~--^

pH

Assay conditions Substrate cont. mM

_____._

suiphate,

DETERMINED

Dodgson and Spencer, rg54

1

AS

5

._“__.

URIXE

= P-acetophenyl

ASD

NPS

-

Substrate

.lPS

SERUM

sulphate,

BLOOD

Huggins and Smith, 1947

-----

: NPS = p-nitropbenyl

OF

- .-~--_I_-____

A uthovs

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ACTIVITY

THE

Substrate used

1

5

TrZBLE

ul “;;

a.

a

ARYLSULPHATASE

383

ACTIVITY

depends on the age of the subject. In the urine of children, activity is very low. It reaches a maximum at the age 30 to 40 years. Diurnal changes of arylsulphatase activity in blood serum and urine have also been observed. According to Boyland’ the activity is highest at noon. The higher activity of the insoluble arylsulphatase C in the urine of female subis jects as compared to male urinesI and the increased activity during menstruation6 due to the presence of an increased number of epithelial cells in the female urines. After centrifugation, the activity in the female urines was equal to that in the male urines. Increased arylsulphatase activity in blood serum was found during pregnancy 13,15. Besides these physiological changes of the arylsulphatase activity, much more pronounced changes have been observed in pathological cases. As was shown by many diseases are associated with increased excretion of arylsulDzialoszynski12, phatase in urine. In the case of myeloid leukemia a 3o-fold increase was observed. Rather substantial increases were also noted in diseases of the bladder, testes and uterus, in cancer of the breast, or of the prostatic gland, and other cases of cancer. Small deviations from normal activity were found in diseases of the heart, in rheumatism and in Hodgkin’s disease. Uterman et al. l9 have also reported that atherosclerosis caused no substantial increase of arylsulphatase activity in urine. Dzialoszactivity in human blood ynski et al.1B observed 30~50”,/, increase of arylsulphatase serum in diseases such as myeloid leukemia, acute nephritis and pneumonia complicated with pulmonary abscess or infarct, or bronchiectasis. A considerable increase of arylsnlphatase activity in urine was noted by Boyland et a!.? in cases of pulmonary and renal tuberculosis. According to these authors, not all cases of cancer were accompanied by increased urinary arylsulphatase, but in cancer of the bladder such an increase was invariably observed. Boyland et al.20 have shown that urinary arylsulphatase does not hydrolyse sulphuric acid esters of aromatic amines which, when not esterified, have strong carcinogenic properties. ARYLSULPHATASE

ACTIVITY

An‘11 TUMORS

The increased excretion of arylsulphatase in the urine in some cases of tumor, as shown by Boyland et d.7~s and DzialoszynskiL2, points to the tumor as the source of the enzyme. Hug-gins and Smith I4 were first to demonstrate an increased arylsulphatase activity in transplantable sarcoma No. 39 in rats. In the connective tissue and skeletal muscle the activitv was 0.015 and 0.17units per g respectively, while in tumor originating from these tissues, it ranged from 0.8-1.6 units/g. Boyland et c.~l.~~~ examined the activity of arylsulphatase in blood serum and urine of normal people and patients \nith cancer of bladder. They found no increase of the activity in the serum of cancer patients. The activity of arylsulphatase in the urine of healthy people was 0.08 pg and in that of cancer patients 1.2-3.3 ilg of 4-nitrocatechol/ml/h which means up to 4o-fold increase. In some cases of cancer, activity in the urine was lower than in the urine of healthy controls. The activity in the urine was not altered after removal of the tumor. Dzialoszynski and Zawielak 21 examined the influence of transplantable Cracker’s sarcoma on the activity of arylsulphatase in various organs of mice. They showed that the neoplasm did not increase arylsulphatase activity per unit weight C/in. Chim. Acta, 15 (1967) 381-386

DZIALOSZYNSKI,

384

GNIOT-SZULZYCKA

of the organ studied, but caused a substantial increase in the size of the spleen and thereby a relative increase of the activity in the whole organism. In the neoplasm itself, activity was four times higher than in the connective tissue from which the tumor originated. Rutenburg and Seligmannz2 studied arylsulphatase activity in some normal and tumorous organs of man using a histochemical method. Their results are not in agreement with those of the above mentioned workers, since Rutenburg and Seligmann have shown only traces of arylsulphatase activity in the tumor, and considerable activity in normal tissue. The cause of this disagreement may be that Rutenburg and Seligmann used 6-benzyl-z-naphthyl sulphate as substrate, while Dzialoszynski and Zawielak applied z-hydroxy-5-nitrophenyl sulphate. the activity of several enzymes, among which arylsulSpencer 23 examined phatase A, R and C in epithelial cell suspensions from bronchi of non-smokers, smokers and from bronchial carcinoma transplants in hamster cheek pouch. The author found no difference in the arylsulphatase activity in cells derived from the above tissues when activity was related to cell volume. arylsulphatase activity in a Recently, Dzialoszynski et al. 24 have examined number of tumors of the stomach, colon, breast and skin using z-hydroxy-5-nitrophenyl sulphate as substrate. They found a pronounced increase of the enzyme activity in all tumors examined, as compared with the activity of the normal tissues from which the tumors originated. This increase was especially high in skin melanomas (about Cio-fold). In all cases examined, arylsulphatase activity in blood serum did not exceed the normal level. activity in a number Dzialoszynski and ~011.I7 found elevated arylsulphatase of tumors of the genital organs of women. In cancer of the uterus the activity was four times higher than in uterine myoma and eight times higher than in nonmalignant ovarian tumors. No increase of arylsulphatase activity was found in the blood serum of the tumor-bearing women. It may be seen that except Rutenburg and SeligmannZ2 and Spencer23, other workers’“,r7,21,24 have found increased arylsulphatase activity in various tumors as compared with the activity in the parent tissues. activity in blood serum was not elevated in any of the cases examined7,17,24. From the work of Royland et al. 7~ it mav be concluded that the increased excretion of arylsulphatase in urine in malignant disease is not directly connected with the increased activity in the tumor itself, since removal of the tumor does not reduce the activity in the urine. The results obtained by Dzialoszynski and Zawielakzr suggest that an increased excretion of arylsulphatase in urine may be connected in some way with the observed enlargement of the spleen. It may also be supposed that the elevated arylsulphatase activity of tumors is linked with the high metabolic rate of the tumor tissue, as it is known from other sources that organs with high metabolic activity22~26-27 and proliferating cells ~--3o have a characteristically high arylsulphatase activity. INFLUEXCE

OF

VARIOUS

FACTORS

ON

THE

ACTIVITY

OF

Dianzani and Ugazzio 31~ studied the influence troduced intraperitoneally, on the activity of various Clin.

Chim

Acta.

15 (1967)

381~386

ARYLSULPHATASE

of carbon tetrachloride, inenzymes (among them aryl-

ARYLSUPHATASE

ACTIVITY

385

sulphatase A and B) in fatty livers induced by that agent in the rat. The authors found increased specific activity of arylsulphatase in liver homogenates at an advanced stage of poisoning. The activity was also increased in lysosome-rich subfractions 3-4 days after treatment of the animals with carbon tetrachloride. Dzialoszynski et aLz8 found increased activity of arylsulphatase in the healing wound. Surgical trauma, however, did not cause any elevation of arylsulphatase activity in blood serum, but a significant increase was observed in the urine29. Dzialoszynski and ~011.~~introduced P-naphthylamine into the stomach of rabbits and observed the level of arylsulphatase in blood serum. They found that arylsulphatase activity in blood serum increased by 40% within 1-8 days after the poisoning. Bianchis4 found that oestradiol injected into guinea pigs caused an elevation of arylsulphatase activity in the liver. The same author has also shown that arylsulphatase activity in human placenta was lower in toxic pregnancy than in normal pregnancy 35. Glick and Stecklein aa studied the influence of various hormones and stress conditions on the activity of arylsulphatase in the adrenals of rat by histochemical methods. They recorded no substantial change in the activity of the enzyme and its distribution in the adrenal tissue under the influence of hormones as ACTH, desoxycorticosterone and cortisone, and under stress conditions as turpentine abscess or cold. Austin et aL3’ found that in metachromatic leucodystrophy arylsulphatase A activity was low in brain, kidney and liver. The activity of arylsulphatase B, on the other hand, in the same organs was increased in gargoylism. CONCLUDI4G

REMARKS

It may be seen from the above that, although determination of arylsulphatase activity in blood serum or in urine may be helpful in some cases, it cannot as yet be introduced into the clinical laboratory as a routine test. This opinion is based on the fact that arylsulphatase is not organ-specific and an elevation of its activity in blood and urine can be caused by many factors. Whereas normal level of activity in blood and urine may exclude a number of diseases, elevated activity in urine may indicate a malignant disease and elevated activity in blood serum allows to conclude on some inflammatory process. REFERENCES J. KRAWCZY~KI, Postep?, Biochem., 5 (1959) 87, I<. GIHI~KI, Polski Tyged. Lekar., 34 (1961) 1,323. L. TOMASZEWSKI, Postefiy Biochem., 8 (1963) 173. H. WEHR, Postepy Biochem., IO (1964) 405. I. SZUMIEL. Postebv BiochFm.. 8 (19611 ~ _ “I 151. Ii. 4MnfoN AND k.‘H. NEY, Arch. Bzoche%. Biophys., 69 (1957) 178. E. BOYLAND, D. M. WALLACE AND D. C. WILLIAMS, Brit. I. Cancer. q (1955) 62. E. BOYLAND, D. M. WALLACE AND D. C. WILLIAMS, Bioch&. J., 56 (195>j_kXIX. K. S. DODGSON AND B. SPENCER, Biockem. 1.. 56 (1954) XIII. IO K. S.Doocso~ AND B.SPENCER,C~~~. Chim. Acta, I (1956)478. II K. S. DODGSON AND B. SPENCER, Biochem. 1.. 65 (1957) 668. IZ L. M. DZIAEOSZY~KI, Clin. Chim. Acta, 2 (1957)542. I 2 3 4 5 6 7 k 9

Clin. Chim.

Acta,

15 (1967)

381-386

386

DZIALOSZYNSKI,

GNIOT-SZULZYCKA

I3 L,. M. DZIAI,OSZYI+,KI AND A. FRBHLICH, Koczniki Wyzszej Szkol~ Rolniczej w Pozna~iu, (1966) in press. Iq C. HIJGGINS -