1642 The American Journal ofClinical Nutrition 31: SEPTEM BER 1978, pp. 1642-1646. Printed in U.S.A.

Digestive-absorptive function of the
intestinal brush border in uremia1
V. W izemann, D. Ludwig, R. Kuhi, and I. Burgmann
ABSTRACT Amino acid absorption was studied in chronic uremic rats. Intestinal transport of
L-leucine appears to be inhibited with mild uremic intoxication, whereas severe uremia enhances
absorption. Brush border activity of intestinal maltase and disaccharidases is higher in rats with
chronic renal insufficiency. The same holds for -y-glutamyl-transpeptidase activity. Am. J. Clin.
Nut,. 31: 1642-1646, 1978.
Although morphological disorders of the
gastro-intestinal tract in uremia are well doc-
umented (1-3), conificting data have been
reported concerning intestinal sugar and
amino acid transport as well as mucosal di-
saccharidase and dipeptidase activities in
renal failure (2, 4-8). The contradicting re-
sults may be explained by different grades of
uremia and by the experimental design. Spe-
cific digestive enzyme activity had not been
measured in the mucosal brush border where
most of the enzymes are located, but in total
mucosal homogenate.
Following the concept of Craine (9) the
intestinal brush border membrane is a “diges-
live-absorptive surface”, where the enzymes
located at the luminal membrane of the mu-
cosa are involved in the transepitheial trans-
port of sugar and amino acids.
Using a brush border preparation, disac-
charidase and dipeptidase activity of intes-
tinal mucosa was determined in normal and
in 90% nephrectomized rats. Referring to the
hypothesis of Orlowski and M eister (10) that
membrane bound -y-glutamyl transpeptidase
plays a role in amino acid transport the en-
zyme was measured too. In order to estimate
the overall effect of uremia on intestinal
amino acid absorption, a preparation of iso-
lated perfused ileal segments was used.
M ethods
One hundred sixteen m ale rats of a Siv strain (body
weight 338 ± 22 g) were used as experim ental anim als.
Uremia was induced by 90% nephrectomy. About 6 to 8
week&”after the operation, serum urea level was 222 ± 27
mg/ 100 ml (control group 48 ± 8 mg/l00 ml). The
animals had free access to Altromin-diet and water.
The brush border membrane was isolated according
to the method of Hopfer et al. (1 l).2 In the m em brane
fraction, activities of the disaccharidases (lactase, cell
lobiase, saccharase, maltase) were determined by the
method of Dahlqvist (12), the dipeptidases (L-leucyl-L-
leucine, L-methionyl-L-methionine, L-methionyl-L-leu-
cine, L-leucyl-glycine, glycine-L-leucine, L-proline-L-leu-
cine) according to Caspary et al. (14).
The transport studies had been performed with iso-
lated perfused ileal segments of normal and uremic rats.
‘4C-L-leucine was used.
Results
In the brush border preparation, disac-
charidases were enriched 13- to 32-fold corn-
pared to the activity in the total homogenate.
The enrichment of dipeptidases in the mem-
brane fraction was comparatively smaller
(Fig. 1).
The measurements of disaccharidases
showed that chronic renal failure only had an
effect on brush border maltase activity, which
was enhanced (P < 0.01) (Fig. 2).
Five of the six determined dipeptidases
showed a higher activity in the intestinal
brush border of the chronic uremic rats (P
< 0.05, Fig. 3). The same held for -y-GT
activity (Fig. 4).
W hen the membrane fraction was incu-
bated with so called uremic toxins, 2-hydrox-
yphenolic acid and 4-aminobenzoic acid
1 From the Department of Internal M edicine, M cdi-
zinische University, 63 Giessen, Federal Republic of
Germany.
2 The authors thank Dr. H. M urer (M ax-Planck-In-
stitut f#{252}r Biophysik, Frankfurt) for his aid in the isolation
method.

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e n ric h m e n t
3 0
- .
i 1
25
2 0
‘5
0 total hem ogenat. 0 b ru s h b o rd e r m e m b ra n e
c p iio b ia s e
m U/m g p ro ?
2 O0
2 3 0 0
FIG . 1. Enrichm ent of disaccharidases and dipeptidases in the brush border of rat intestine.
2 2 0 0
2 1 0 0
2 0 0 0
1 9 0 0
0 0
3 0 0
2 0 0
1 0 0
0 c o n tro l jre m ic
FIG . 2. D isaccharidase activity in the intestinal
brush border.
( l0 M /liter), the activity of all dipeptidases
w as inhibited by 24 to 36% . The addition of
urea (300 m g/lO O m l), creatinine (20 m g/l00
m l), and m ethylguanidine (l0 M /liter) had
A M IN O A C ID A B SO R PTIO N IN C H R O N IC U R EM IA 1643
r’ailase lactose s a c c h a rin i no effect on enzym e activity. D isaccharidases
w ere not influenced by either of the toxins.
In the transport studies the absorption of
L-leucine appeared to be depressed in rats
suffering from a m oderate chronic renal in-
sufficiency. In end stage urem ia, L-leucine
absorption seem ed to be enhanced (Fig. 5).
D iscussion
In contrast to the fm ding of dim inished
dipeptidase activities in the w hole hom oge-
nate of m ucosal biopsy m aterial from urem ic
patients (2, 6, 7), anim al experiences indicate
no alteration of enzym e activity in renal fail-
ure (w ith the exception of leucine-proline
dipeptidase, w hich w as enhanced) (5). In the
present study, m em brane bound intestinal
dipeptidases w ere significantly increased in
the urem ic anim als. A possible role of brush
border located -y-G T is indicated in Figure 4.
A ccording to the hypothesis of O rlow ski and
M eister (10) the enzym e transfers the -y-glu-
tam yl group of glutathione to an acceptor,
w hich can be an am ino acid or a sm all pep-
tide. C orresponding to the dipeptidase results,
-y-G T activity is higher in the urem ic brush
border.
Sm all m olecular urem ic toxins (14) had no
influence on brush border enzym e activity
w ith the exception of phenolic com pounds
and arom atic am ines. The concentration of

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9500
m U/m g p ro t
9000
8500
0 c o n t r o l
u r e mi c
l# {1 4 9 }u -lt’u m e t-m e t met-ieu leu-g(y g ly -ie u p . ro i-ie u
FIG . 3. B rush border activity of dipeptidases.
1644 W IZEM A N N ET A L.
8 0 0 0
7 5 0 0
7 0 0 0 # {1 4 9 }
6 5 0 0
6 0 0 0
5500

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m g pro f
4 0 0
3 0 0 # {1 4 9 }
2 0 0
1 0 0
-I
0 c o ntro l
0 ure m ic
active tra ns po rt o f L- le uc m ne in the is o la te d ile um
( ; - urea 6 2 8 m g . /.
7
I
I
I
I
I
L
.ig Ieucmne
r)gprot amin
2 . 5
2.0-
1 5
1 0 -
0 5-
0#{149}
1 _ -‘A t .
I ./‘
0
ure a 2 lOm g ’/.
urea 154 m g ’Y.
I
e quilibra tio n
0
1 T
25 35 45 55 65 75 85 mm
FIG . 5. Absorption of L-leucine at different grades of urem ic intoxication in rats.
A M IN O A C ID A B SO R PTIO N IN C H R O N IC U R EM IA 1645
1 ’ -Glu-Cys -Gly# {1 4 9 }a c c e pto r
(g lura thio ne )
r-Glu -a c c e pto r. Cys - Gly
FIG . 4. -y-G T activity. The equation gives an outline
of the concept of O rlow ski and M eister (10).
the tw o toxins w as about 10-fold higher than
in the serum of urem ic patients. The rat
intestinal absorption of leucine show ed a bi-
phasic pattern in chronic renal failure, op-
posite to the behavior of the exocrine pan-
creas (15). A t a m oderate degree of urem ic
intoxication, am ino acid resorption appears
to be inhibited, w hereas in term inal renal
failure, transport is enhanced. In experim en-
tal diabetes of rats a sim ilar constellation of
an increased digestive and transport function
of the intestinal m ucosa w as found (16).
There are no data available on intestinal
am ino acid absorption in urem ic patients.
B ased on the fact that urem ics can m aintain
a neutral or even positive nitrogen balance
under a strictly protein-restricted diet one can
assum e that am ino acid absorption is either
undisturbed or enhanced. O ur rat experi-
m ents support this hypothesis.
0/
7 - . . urea52mg/.

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1 6 4 6 W IZ EM A N N ET A L .
R ef erences
I. CA S T R UP, H. J., U. L OHR S A N D H. EDER. A ut or a-
diographische und histochem ische Untersuchungen
z ur Entstehung der sogenannten Enterocolitis. V ir-
chow s A rch. A bt. A Pathol. A nat. 340: 357, 1970.
2. DEN N EB ER G, T ., T . L I NDBERG, N . BERG A ND A .
DA H L QV I ST . M or phol ogy, di pept i dases and di sac-
charidases of sm all intestinal m ucosa in chronic renal
f ailure. A cta M ed. S cand. 195: 465, 1974.
3. M A SON, E. E. Gastrointestinal lesions occurring in
urem ia. A nn. Internal. M ed. 37: 96, 1952.
4. GRI M M EL , K ., A N D H. K A SPER. Car bohydr at e, pr o-
tein and f at absorption in chronic uraem ia. In: Di-
aly sis, T ransplantation, N ephrology , edited by B . H.
B . R obinson. Pitm an M edical, 1976, pp. 566.
5. GRI M M EL , K ., S . BONGA RT Z A ND H. K A S PER . T he
activ ity of intestinal disaccharidases and dipepti-
dases in rats w ith chronic renal insuf f iciency . A cta
Hepato-Gastroenterol. 21: 383, 1974.
6. M A DZ A R OV A -N OHEJL OV A , I. S m all bow el disac-
char i dase act i vi t y in hy peraz otem ic states. Prague:
V IIIth International Congress of Gastroenterology ,
July 7-15, Prague, 1968.
7. M CNA I R, A ., J. OL S EN . Disaccharidase activ ity in
chronic renal f ailure. A cta M ed. S cand. 195: 93,
1974.
8. POIN T N ER , H., U. FL EGEL , C. BA Y ER A ND P.
SCH M I DT . A nwendung des T r ehal ase-T ol er anzt est s
z ur Untersuchung der D#{ 252} nndarm resorption bei Pa-
tienten unter chronischer H#{ 228} m odilay se. W ien. K lin.
W ochochr. 86: 691, 1974.
9. CRA NE, R . K . S tructural and f unctional organiz ation
of an epi t hel i al cel l brush border. In: Intracellular
transport, edited by K . B . W arren. N ew Y ork : A ca-
dem ic Press, 1967, pp. 71.
10. OR L OW S K I, M ., A N D A . M EI ST ER. T he y -glutam y l
cy cle: a possible transport sy stem f or am ino acids.
Proc. N atl. A cad. S ci. US A 67, 1248, 1970.
11. H OPFER, U., K . N EL S ON , J. PER OT rO A N D K . J.
IS S EL B A CHER . Glucose transport in isolated brush
border m em branes f rom rat sm all intestine. J. B iol.
Chem . 248: 25, 1973.
12. DA H L QV I ST , A . A m ethod f or assay of intestinal
disaccharidases. A nal. B iochem . 7: 18, 1964.
13. CA SPA RY , W . F. Intestinale Peptidhy drolasenak tiv it
at in m enschlichem D#{ 252} nndarm biopsiem aterial-
eine einf ache B estim m ungsm ethode. K lin. W o-
chechr . 52: 341, 1974.
14. H I CK S, J., D. Y OUN G A N D I. W OOT ON . T he ef f ect
of ur emi c blood constituents on certain cerebral
enz y m es. Clin. Chim . A cta 9: 228, 1964.
15. W IZ EM A N N , V . Ex ocrine pancreatic f unction in
chronic renal f ailure. I n: Di al ysi s, T r anspl ant at i on,
N ephrology , edited by B . H. B . R obinson. Pitm an
M edical, 1976, pp. 585.
16. CA SPA RY , W . F., A . M . RH EI N A ND W . CREUT Z -
FEL DT . I ncr ease of di gest i ve and t r anspor t f unct i ons
of i nt est i nal mucosa f r om st r ept ozot ozi n-di abet i c
rats. Europ. J. Clin. Inv est. 2: 278, 1972.
17. GUL Y A SSA Y , P., J. PET ER S A N D P. S CHOEN FEL D.
Urem ia. S tuttgart: T hiem e V erlag, 1972, p. 163.
18. K OPPL E, J. D., A N D M . E. S W EN DS EID. A m ino acid
and k eto acid diets f or therapy in renal f ailure.
N ephron 80: 1, 1977.

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