Methylprednisolone-Induced Hypertension

Role for the Autonomic and Renin Angiotensin Systems

OSVALDO KOHLMANN JR., M.D., ARTUR B. RIBEIRO, M.D., ODAIR MARSON, M.D.,

MANOEL A. SARAGO£A, M.D., AND OSWALDO L. RAMOS, M.D.

SUMMARY The role of the autonomic nervous system (ANS) in the pathogenesis of hypertension induced
by methylprednisolone (20 mg/kg/week subcutaneously) was studied in rats before and during chronic renin
angiotensin system (RAS) blockade with captopril (20 mg/kg/erery 8 hrs by mouth). Sympathetic nervous
system (SNS) blockade was accomplished by the intravenous (i.v.) administration of propranolol (0.20
mg/lOOg) plus phentolamine (1.25 mg/lOOg/i.r.) and ganglionlc (G) blockade by the use of pentollnium tar-
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tarate (0.5 mg/100g/i.v.). After 4 weeks, methylprednlsolone-treated animals'showed significant decreases in

mean arterial pressure (MAP) with both SNS (-34 ± 2 ram Hg) and G (-56 ± 3 mm Hg) blockades; during
chronic RAS blockade, even greater falls in MAP were observed (SNS = -43 ± 2 mm Hg and G = -75 ± 3
mm Hg). Nevertheless, for both groups the levels of MAP obtained during SNS and G blockades were higher
than those observed in their control groups. At the end of second week, however, in captopril-treated hyper-
tensive rats the values of MAP obtained during ANS blockade were lower than those observed in the control
group. An increased responsiveness to exogenous administration of norepinephrine (NE) was observed in
animals receiving methylprednisolone and captopril. It is concluded that methylprendisolone hypertension in
the rat may be initially explained by activation of RAS and ANS. At later phases, a third mechanism has to be
postulated to explain the hypertensive state. (Hypertension 3 (suppl II): II-107—11-111, 1981)

KEY WORDS * glucocorticoid-hypertension • autonomic nervous system •

renin angiotensin system • norepinephrine responsiveness • captopril • saralasin

S EVERAL mechanisms have been proposed to

explain the pathogenesis of excess glucocorti-
coid hypertension. They include: salt reten-
tion,1 activation of the renin angiotensin system, 1 ''
increased plasma and extracellular volumes,4 and al-
terations in various components of the autonomic ner-
vous system (ANS).6"7
We have recently confirmed that hypertension in-
duced by methylprednisolone in the rat is accom-
panied by activation of the renin-angiotensin system.'
However, this hypertension may occur even during
chronic renin angiotensin system blockade with cap-
topril," leading to the conclusion that other mech-
anism^) may participate in the pathogenesis of this
hypertension. In this study we tested the role of the
ANS in hypertension induced by methylprednisolone
in the rat. Special emphasis is placed upon the interac-
tion of the autonomic and renin angiotensin systems in
this type of experimental hypertension.
Methods
Induction of the Hypertensive State
Male Wistar rats receiving regular rat chow and tap
water were used. Four groups of rats were prepared: 1)
Group M (n = 20), receiving a long-acting prepara-
tion of soluble methylprednisolone (Depo-Medrol,
Upjohn) at a dose of 20 mg/kg/week subcutaneously;
2) Group M + C (n = 20), receiving methyl-
prednisolone as in Group M plus concomitant cap-
topril (Squibb and Sons') at a dose of 20 mg/kg/every
8 hours by gavage; 3) Group N (n = 20), receiving dis-
tilled water by the same route and same frequency as
Group M; 4) Group N + C (n = 20), receiving dis-
tilled water as Group N plus concomitant daily cap-
topril given as in Group M + C. All groups were
followed for 4 weeks, and tail arterial pressure was
registered every week using a tail microphonic
method' in unanesthetized animals.

From the Department of Medicine, Division of Nephrology,
Escola Paulista de Mcdicina, SSo Paulo, Brazil
Dr Ribeiro is an Established Investigator from the Brazilian
Council for the Development of Science and Technology (CNPq)
Address for reprints Artur B. Ribeiro, M D, Associate
Professor of Medicine, Division of Nephrology, Escola Paulista de
Medicina, Rua Botucatu, 720-04023 S3o Paulo, Brazil
11-107
Pharmacological Blockade of the Sympathetic Nervous
System (SNS)
At the end of the fourth week, 10 animals of each
experimental group (M; M + C; N, and N + C) had
catheters placed into the carotid artery and jugular
 11-108 PROCEEDINGS/INTERAMERICAN SOCIETY
vein under ether anesthesia. On awakening, the rats'
mean arterial pressure (MAP) was continuously
recorded with a Beckman physiograph, and the ani-
mals were kept unrestrained with free access to water
and food. After a 2-hour control period, blockade of
sympathetic nervous system (SNS) was induced by in-
travenous administration of propranolol (Inderal,
Ayerst Laboratories) at a dose of 0.20 mg/100 g body
weight, followed by phentolamine (Regitine, Ciba
Geigy Corporation) at a dose of 1.25 mg/100 g body
weight/i.v. The MAP was recorded for 60 minutes
after the initial injection. Results are expressed by
differences in MAP values before propranolol and 30
minutes after phentolamine administration. Identical
procedures as described above were used in animals of
Groups M + C (n = 12) and N + C (n = 12) studied
at the second week.
Ganglionic Blockade and Vascular Responsiveness to
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Norepinephrine (NE)
Four weeks after the initiation of methylpred-
nisolone administration, 10 animals of each group (M,
M + C, N, and N + C) were prepared, as described
above. After a 30-minute control period, norepineph-
rine (NE) (Noradrenaline, ByK Laboratories) was ad-
ministered at increasing doses of 50, 100, and 200
ng/100 g body weight, i.v. Each injection was given
after the MAP had stabilized following the previous
injection, and each dose was given twice. The effect of
NE upon MAP is represented by the mean of the two
maximal elevations observed for each dose used.
Following a recovery period from the last dose of NE,
all animals underwent ganglionic blockade by the ad-
ministration of pentolinium tartarate (Ansolysen,
Wyeth Laboratories) given as a single intravenous
dose of 0.5 mg/100 g body weight. The MAP was
registered continuously, and after stabilization,
responses to NE were evaluated as before pento-
linium. Vascular responsiveness to NE was also tested
in eight animals of Group M + C and N + C studied
at the second week.
Results are expressed as mean ± standard error of
the mean. Statistical analysis were obtained by Stu-
dent / test or analysis of variance when appropriate.
Results
Table 1 shows the values of tail arterial pressure
(TAP) observed in the four groups studied. As can
be seen, in animals receiving only methylpredniso-
lone TAP was significantly elevated by 1 week, and
reached a maximal elevation of 41 ± 1 mm Hg at 4
weeks. In the group receiving captopril methyl-
prednisolone (M + C), a progressive rise in TAP was
observed only after the first week, and a maximal
change of 24 ± 1 mm Hg was observed at the fourth
week. These values were significantly lower than those
observed in Group M (p < 0.01). No significant
changes in TAP were noted in Group N, whereas a
small but statistically significant decrease (ATAP =
- 9 ± 1 mm Hg, p < 0.01) was observed in Group N
+ C at the fourth week.
S U P P II, HYPERTENSION, VOL 3, N o 6, N O V / D E C , 1981
TABLE 1. Tail Arterial Pressure (in mm Hg) in groups M,
M+C,NandN+C
Weeks
Groups Control 1 2 3 4
M 114 ± 1 126 ±1* 138 ± 1 * 147 ± 1* 155 ± 1*
M + C 116 ± 1 119 ± 1 122 ±1** 134 ± 1** 140 ±1**
N 114 ± 1 116 ± 1 117 ± 1 117 ± 1 116 ± 1
N + C 116± 1 114 ± 1 110 ± If 108 ± IT 107 ± IT
•p < 0.01 compared to groups M + C, N and N + C.
**p < 0.01 compared to group N and N + C.
tp < 0.01 compared to group N.
Absolute values of direct MAP, as well as changes
in MAP induced by SNS and ganglionic blockade, are
summarized in table 2 and figure 1 respectively. It is
apparent that both SNS and ganglionic blockade
evoked significant drops in MAP in all groups. During
SNS blockade, reductions in MAP of methyl-
prednisolone-treated groups (M = — 34 ± 2 mm Hg
and M + C = —43 ± 2 mm Hg) were significantly
greater (p < 0.01) than those observed in the control
groups (N = - 1 5 ± 2 mm Hg and N + C = - 2 4 ± 1
mm Hg). Similar but greater decreases in MAP were
observed during ganglionic blockade: M = —56 ± 3
mm Hg; M + C = - 7 5 ± 3 mm Hg; N = - 3 4 ± 2
mm Hg; and N + C = - 4 8 ± 2 mm Hg. It is also
noteworthy that in the captopril-treated groups (M +
C and N + C) changes in MAP induced by both
blockades were significantly greater than those ob-
served in the groups that did not receive that drug (see
figure 1). Furthermore, during autonomic nervous
system blockade MAP values in the M + C group
TABLE 2. Mean Arterial Pressure (MAP) m mm Hg Before
and During Sympathetic Nervous System (SNS, Proprano-
lol + Phentolamine) and Ganglionic (Pentolinium Tartar-
ate) Blockade m Groups M, M + C, N and N + C Studied at
the 4th week (X ± SEM)
Groups Before SNS Ganglionic
M 154 ±2* 118 ±2* 100 ±4*
M+C 136 ± IT 93±2t 60±3T
N 115 ±3TT 100±3TT 81 ±2TT
N+C 98 ± 1 73 ± 2 50±l
*p < 0.01 — compared to groups M + C, N and N + C.
tp < 0.01 — compared to group N + C.
ftp < 0.01 — compared to group N + C.
TABLE 3. Mean Arterial Pressure (MAP) m mm Hg Before
and During Sympathetic Nervous System (SNS, Proprano-
lol + Phentolamine) and Ganglionic (Pentolinium Tartar-
ate) Blockade in Groups M+C, and N+C Studied at the
2nd Week (X ± SEM)
Before SNS Ganglionic
Groups
M+C 112 ± 1 * 76 ±2* 50 ±3*
N+C 98 ± 2 85±3 61 ± 2
*p < 0.01 compared to group N + C.
 METHYLPREDNISOLONE-INDUCED HYPERTENSION/Kohlmann et al. 11-109

fiMAP 0 AMAP
(mriHg}
(mmHg)
-10 -
-10 -
-20 -
N+C
N+C -20 -
-30 -

-40 - -30 -
M+C
-50 - M+C
-40 -
SNS
-60 -
D
0 GANGLIONIC -50 •
-70 -
Q SNS
-80 - GANGLIONIC
-60 -
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FIGURE 1. Decreases in mean arterial pressure (AMAP),
in mm Hg, during sympathetic nervous system (propranolol
+ phentolamine) and ganglionic (pentolinium tartarate)
blockades in Groups M, M + C, N and N + C studied at
fourth week. *p < 0.01 compared to Group M, **p < 0 01
compared to Group N
J
-70
FIGURE 2. Decreases in mean arterial pressure (AMAP),
in mm Hg, during sympathetic nervous system (propranolol
+ phentolamine) and ganglionic (pentolinium tartarate)
blockades in Groups M + C and N+C studied at second
week *p < 0 01 compared to Group N + C.

reached levels that were significantly lower than those

observed in the M group, and similar to those of the
N group. However, MAP in the M + C group re-
mained significantly higher than in the N + C group
(see table 2). AMAP
Values of MAP in the groups studied at the second (mmHg)
week (M + C and N + C) before and during SNS and 120 -
BEFORE
ganglionic blockade are shown in table 3 and figure 2. DURING GANGLIONIC * •

Basal MAP values in M + C group were significantly BLOCKADE

higher than those of N + C. Also, both blockade 100 -
procedures resulted in evident decreases in MAP *T*
values for both groups. However, drops in MAP were
of greater magnitude in M + C animals ( - 3 2 ± 2 and
—59 ± 2 mm Hg, figure 2) as compared to those ob-
80 -
]lc
tained in Group N + C ( - 1 3 ± 1 mm Hg and - 3 6 ±
** ...-'"' . ', •
2 mm Hg) {p < 0.01 for both procedures). Thus, dur- 60 -
ing both blockades, MAP was lower in Group M + C I . - - ' ' . - - ' • ' ^ , - ' '

as compared to Group N + C. 1' ___J^ + C

Figure 3 shows the increments in MAP obtained TM
40 • =S3 N
with NE injections before and during ganglionic - ^ ^ N+C
blockade. It is evident that the NE-induced incre-
ments in MAP were greatly enhanced by ganglionic 20 •
?
blockade. Both before and during ganglionic block-
ade, elevations in MAP induced by NE were greater in
Group M + C as compared to the other groups (p <
0.05, M + C vs M or N or N + C).
50 100 200 NE(ng/IOOg)
Figure 4 shows elevations of MAP induced by NE
before and during ganglionic blockade in Groups M +
C and N + C studied at the end of second week. For FIGURE 3. Mean arterial pressure elevations (AMAP)
all but the first NE dose tested, elevations in MAP for with increasing doses of norepinephrtne (NE) before and
both groups were significantly greater during gan- during ganglionic blockade at the fourth week. *p < 0 05
glionic blockade. Again, greater elevations in Group compared to Groups M, N, and N+C before ganglionic
M + C were observed before and during ganglionic blockade **p < 0 05 compared to Groups M, N, and N +
blockade for all three NE doses tested. C during ganglionic blockade.
 11-110 PROCEEDINGS/INTERAMERICAN SOCIETY SUPP II, HYPERTENSION, VOL 3, N o 6, N O V / D E C , 1981

A MAP Thus, methylprednisolone hypertension cannot be ex-

mmHg)
plained only by hyperactivity of the ANS. These
eo - results agree with a previous report from our labora-
tory which documented a role for the RAS in the
BEFORE maintenance of this type of hypertension.' In the
DURING GANGLIONIC group receiving captopril and methylprednisolone (M
100 - BLOCKADE + C), ANS blockade caused decreases in MAP that
V M+C were greater than those of Group M. Also, the levels
of M A P attained during ganglionic and SNS
60 - y blockades were lower in M + C as compared to M.
This fact helps to corroborate the hypothesis of a
N+C simultaneous activation of the RAS and SNS in the
•• methylprednisolone-induced hypertension. However,
60 - the fact that, during ANS blockade, the blood pres-
^y M+C sure levels of the captopril-treated group (M + C)
were still higher than those of its appropriate control
(N + C) suggests that a third factor, in addition to
40 - N+C
ANS and RAS, must be involved to explain the hy-
•P pertension following methylprednisolone adminis-
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20 -
r"
^-^^^
50 100 200 NE
(ng/IOOg)
FIGURE 4. Mean arterial pressure elevations (LMAP)
with increasing doses of norepinephnne (NE) before and
during ganglionic blockade at the second week *p < 0 05
compared to Group N + C before ganglionic blockade. **p
< 0 05 compared to Group N + C during ganglionic
blockade.
Discussion
A role for the renin angiotensin system (RAS) in
the pathogenesis of methylprednisolone hypertension
in the rat has been previously demonstrated.*• *• *
However, we have reported previously that RAS
blockade with captopril does not prevent the develop-
ment of this type of hypertension." This observation is
confirmed in the present work, since elevations in arte-
rial pressure were evident at the fourth week in the
group treated with methylprednisolone and captopril
(M + C). Thus, other mechanism(s) must have come
into play to explain the elevations of MAP during
chronic RAS blockade.
Our results clearly show that administration of
methylprednisolone for 4 weeks causes hypertension
accompanied by an increase in SNS activity. Accord-
ingly with both pharmacological blockade procedures
used, significantly greater drops were observed in
methylprednisolone-treated rats. In the group of
animals receiving only methylprednisolone (M), both
blockades elicited significant drops in MAP; however,
the levels of MAP attained were still greater than
those observed in the appropriate control (Group N).
tration.
In contrast, at the second week of glucocorticoid
administration, both SNS and ganglionic blockades
completely reversed elevations of MAP observed in
Group M + C. Actually, during these blockades,
MAP levels were even lower in Group M + C com-
pared to those of Group N + C. Thus, it is apparent
that in the earlier phases of methylprednisolone ad-
ministration the elevations in blood pressure may be
explained by simultaneous activation of the RAS and
ANS only.
Whereas the role of the renin-angiotensin system in
this type of hypertension is well established,2' ' • ' the
effect of glucocorticoid administration on SNS func-
tion remains controversial. 1 ' 6 ' '• '• 10 Some others have
suggested alterations in tissue metabolism of cate-
cholamines" and others, an increase in vascular sensi-
tivity to exogenous NE.' 1 7 In our work we observed
that methylprednisolone alone did not significantly
alter vascular responsiveness to the three doses of ex-
ogenous norepinephrine used, since MAP elevations
were similar in Group M and N both before and dur-
ing ganglionic blockade. Comparable results were pre-
viously reported in the same species studied at 2
weeks.2 However, when methylprednisolone was asso-
ciated with captopril (Group M + C), a significant in-
crease in responsiveness to these doses of NE was ob-
served before and particularly during ganglionic
blockade. This was observed already at the second and
was still present at the fourth week. This increased
responsiveness to NE cannot be attributed to captopril
alone since the increments in blood pressure observed
in Group N + C are not different from those of Group
N (fig. 3). Thus, we could observe an increased respon-
siveness to three exogenously administered doses of
NE only with the association of methylprednisolone
and captopril. Although interpretation of these results
is difficult, one might speculate that a diminished
endogenous liberation of catecholamines induced by
the diminution of angiotensin II caused by captopril11
could unmask the reported hypersensitivity of
adrenergic receptors caused by corticosteroid.7 The
enhanced responsiveness observed in the Group
 METHYLPREDNISOLONE-INDUCED HYPERTENSION/Kohlmann el al.
M + C after ganglionic blockade could help to sup-
port this hypothesis.
In conclusion, it is possible to detect the participa-
tion of both the renin-angiotensin system and the
autonomic nervous system in the pathogenesis of glu-
cocorticoid hypertension. In the early stages (second
week) of hypertension, these two factors seem to be
responsible for the elevated blood pressure. Subse-
quently (fourth week) a third factor must be
postulated to explain the raised arterial pressure. The
apparent role of the ANS hyperactivity in this type of
hypertension may be, at least in part, explained by an
increased responsiveness to catecholamines.
References
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 Methylprednisolone-induced hypertension. Role for the autonomic and renin angiotensin
systems.
O Kohlmann, Jr, A B Ribeiro, O Marson, M A Saragoça and O L Ramos

Hypertension. 1981;3:II-107
doi: 10.1161/01.HYP.3.6_Pt_2.II-107
Hypertension is published by the American Heart Association, 7272 Greenville Avenue, Dallas, TX 75231
Copyright © 1981 American Heart Association, Inc. All rights reserved.
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