JKAU: Sci.. vol. 9,pp. 31-48 (14"7 A.H./1997 A.D.)
The Ability of Perindopril of Modulate Behavioural, Neuroendocrinal and Testicular Profiles in Stressed Rats
MANSOUR A. AL-HAZMI*, OMNIANAYEL**, OSAMA SOROUR*** and NADIA EL-BANNA**
*Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah. Saudi Arabia
**Pharmacology, Drug Toxicology, and ***Dermatology & Venerology Departments, Faculty of Medicine, Alexandria University, Egypt
ABSTRAc.'T. The study was conducted on 42 adult male Wistar rats, receiving a daily IP injection of: I) 0.2 ml saline, in control group (n = 6). 2) 0.2 mI sa- line in stress group (5-GP) (n = 18). 3) 400 Ilg/kg perindopril, in perindopril pretreatment stress group (P-P 5-Gp) (n = 18). After 3 weeks, animals in group 2 & 3 were each equally subdivided and subjected to short term modalities of stress: Heat & Light (H&L-5), (n = 6), Cold (C-5), (n = 6) and immobilization
& Immersion (1&1-5), (n = 6). The sequelae of these were firstly assessed on some behavioural activities, then on some neuroendocrinal responses and sub- sidiary testicular histopathological changes, instantaneously after decapitation.
Results revealed that P-P H&L-5 prevented the significant changes in adre- nocorticotrophic hormone (ACTH), testosterone and non-social investigations' and significantly decreased, but to lesser extent, the elevation in norepine- phrine (\liE), yet did not prevent the significant alterations in dopamine (DA), cortisol and displacement, induced by H&L-5, in comparison to control. Mean- while, this pretreatment increased significantly the social investigations and 5HT in comparison to control. On the other hand, P-P C-5 prevented the sig- nificant changes in NE and threat induced by C-5, yet it significantly increased social investigations, in comparison to control. However, P-P 1&1-5 prevented the significant changes in prolactin (PRL) and non social investigations and significantly decreased, to a lesser extent, the changes in NE, DA, ACTH, cor- tisol, testosterone and defence, but pertained the significant elevation in 5HT, induced by 1&1-5, in comparison to control. Meanwhile, this pretreatment sig- nificantly increased social investigations and decreased displacement in com- parison to control. No testicular histopathological changes were inflicted by 5- Gps or modulated by P-P5 Gps.
Introduction
Studied probing in the impacts of stress on psyche and soma are still a dilemma without solid links. On epidemiological basis, the role of stress as a segregated etiological factor
31
M.A. AI-Haznli et al.
32
in different psychological cardiovascular, endocrinal and metabolic disorders and dis- turbances have long been raised[I-3]. However, when translated into biochemical terms;
the psychoemotional provocation aroused by wear and tear insults, that are evoked by different threatening physical and/or psychological stresses appear uptill now to intricate nature[2,4]. Though, a common non specific hypothalamic-pituitary-adrenocor- ticomedullary release and sympathetic nerve terminal stimulation, i.e., what is termed neuroendocrinal response to stress, has long been coined[2,S]. But, whether or not the var- ied stressors, are able to provoke the release of the same synaptic and humoral com- ponents of this response and with the same intensity, is still argued and nee,ds to be ex- plained[4,6,7]. Furthermore, only few studies have demonstrated an etiopathophysio- logical link between all forestated varients and the intersecting hypothalamic-pituitary- testicular axis, to influence male fertility, a point that needs to be thproughly highlighted.
The present study, thus focuses on the impacts on three modalities of stress, linking their related behavioural alterations to the probable relevant simultaneous fluctuations in brain neurotransmitters, pituitary and steroid hormones and histopathological altera- tions in the testis of male rats.
In addition, the study also examines a drug such a perindopril [P] (Coversyl, 'Servier L ill. French[8] a novel angiotensin converting enzyme inhibitor, classically admin- istered to control hypertension [where stress per se plays a relevant etiological pre- cipitating fac.tor][3]. It questions; whether or not this drug can favourably ameliorate the varied impacts of stress tackled in this study, as efficiently as, when controlling the hy- pertensive outcvme of stress[8]. If so is the case, then recognizing drugs for use that can strike at multiple levels to curtail tpe hazardous outcomes of stress on psyche and soma would be an important target to achieve.
Experimental Procedures
42 adult male Wistar rats [170-200 g] were reared under contrplled conditions of tem- perature and reversal light cycle. Food and water were added ad libitum. After one month of being acclimatized, ~nimals were then divided into three groups which re- ceived a daily IP injection of (a) 0.2 ml saline in the control group (n = 6), (b) 0.2 ml s~- line in the'stress group [S-Gp] (n = 18), and (c) 400~g/kg P(8) [Coversyl -Servier LTD.
French} in perindopril treatment stress group [P-P S-Gp] (n = 18). Animals were kept under the same standard conditions fQr three more weeks after which those of groups (2) arid (3) were further equally subdivided into three relevant subgroups, each of which being subjected to one of the short term modalities of stress experimented namely:
Heat and Light'Stress [H&L-SJ; where the top of the cage was continuously elu- minated for 36hr by 200V lamp; thus breaking the light cycle and confering therinal in- sult throughout[9].
Cold Stress [C-SJ; where rats were refrigerated for 12hr at 4°C[IO].
Immobilization and Immersion Stress [I&I-SJ; where the fore and hind limbs of
each rat were tied up to and locked i~ a small metal cage, that was immersed in cold wa-
ter 4°C for 24hr, while sparing its head out[ll].
The Abilif)' of Perindopril to"
33 Ethological Behavioural Assessments
Once stress experimentations were terminated, each animal from the three studied groups was placed into a special partitional cage against its standard opponent[12] and the broad behavioural categories, i.e. social, non-social investigations, attack, threat, de- fence and displacement as well as the number of actual attacks were all recorded over 900 seconds using coloured video camera[13]. The tapes were then analysed for al- locating the time consumed in each behavioural category[12,13]. These data were sta- tistically analysed using Kruskal-Wallis test with appropriate comparison being made using Mann-Whitney "U" test[14].
Analytical Biochemical Estimates
After terminating the previous testing, each animal was decapitated, the blood was in- stantaneously collected and the brain was simultaneously frozen in liquid nitrogen once dissected. These samples were used to determine:
Neurotransmitters: where the whole brain was weighted and homogenised to de- termine norepinephrine (NE), dopamine (DA) and serotonin (5HT) by the fluorometric
micromethod[15] using an Aminco Bowman fluorospectroph6tomete~.
Hormones: where serum ACTH, PRL and testosterone levels were determined by a RIA technique[16-18] and serum cortisol level was determined by an ELISA technique[19].
All biochemical data were statistically analysed using the student (t) test for unpaired comparison[20]
Histopathological Changes
The testes were removed from all studied animals, the epididymis was dissected out and the testicular tissue was impregnated in Buan solution then Formalin, processed, sec- tioned and stained by H&E to detect any structural changes least they have developed{21].
Results
Behavioural Changes; Demonstrated in Table 1 and Figure (1)
The time allocated to non social investigation was significantly prolonged and that to displacement was significantly by H&L-S in comparison to control. P-P H&L-S, sig- nificantly shortened the time advocated to threat in comparison to its relevant H&L-S and pertained that of displacement shorted in comparison to control. The tim~ con- sumed in threat was significantly decreased in C-S and I&I-S, while that of non'social investigation was significantly shorted and that of defence attempt was significantly prolonged only in I&I-S all in comparison to control. P-P I&I-S significantly shortened time allocated to non-social investigation, defence and displacement in comparison to relevant I&I-S, the latter attempt, being also significantly shortened in comparison to control. In all P-P S-Gps, the time consumed in social investigation was significantly prolonged whether in comparison to control or to their respective S-Gps.
34 M.A. AI-Haznu el al.
Whole Brain Contents of Monamines Demonstrated in Table 2 and Figure (2) NE and DA were significantly increased by H&L-S and tended to decrease by P-P H&L-S, though still being considered; significantly elevated, when compared to control.
However, NE induced decrease, was significant when comparing P-P H&L-S to rel- evant H&L-S alone. On the contrary, this drug regimen induced a significant increase in 5HT in comparison to control. In C-S only NE was significantly elevated and P-P to C- S protected against the rise, but meanwhile, induced a significant elevation in 5HT in comparison to control. In I&I-S all studied arnines were significantly elevated while 5HT was pertained. significantly elevated but NE and DA tended to decreased by P-P I&I-S though were still co.nsidered elevated in comparison to control.
TABLE I. Ethological behavioural changes in rats subjected to three different modalities of stress per se or after three weeks pretreatment with perindopril (400 mg/kg) daily (P. (Value are expressed as Median &
Range and compared by using Mann-Whitney "U" test).
Time allocated to ethological changes in seconds Behavioural
assessment
studied Gps Displace-
ment
No. of attacks
Attack Threat Defence
Coll/rol Gp.
(0=6) Stress Gps.
a) H&L-S (0=6) b) C-S
(0=6) c) I&I-S
(0 =6) p.p S- Gps.
a) pop H&L-S (0=6) b) pop C-S
(0 =6) c) pop I&I-S
(0=6)
Social investigation
202.5 [65 -280]
Non social investigation
280.0 [150 -395]
0.0 [0.0-0.0)
25 [10-40]
0.0 [0.0-10]
13.0 [35 -215]
0.0 [0.0-0.0]
54.0 [44 -200]
189.0 [68 -489]
115.0 [75 -150]
410*
{228 -488]
289.0 [105:475]
65.5***
[45 -160J 0.0 [0.0 -0.0]
0.0 [0.0-0.0]
0.0 [0.0 -0.0]
36.0 [0.0-110.0]
0.0***
[0.0-16.0]
0.0***
[0.0-0.0]
0.0
[0.0-2.0]
0.0
[0.0-0.0]
215.0...
[150 -265]
I
30.0I [24-108]
57:0 [0.0-122]
215.0 [105 -240]
0.0 [0.0 -8.0J
0.0 [O.O-O.OJ
0.0 i [O.O-O.OJ 569.0**+++
[254 -781]
607.5**+++
[477 -769]
645.0***+++
[465.719]
269.5 [68 -621]
376.5 [53 -357]
239,0+++
[170-413]
0.0 [0.0-0.0]
0.0 [0.0-0.0]
0.0 [0.0 -0.00]
0.0++
[0.0-0.0]
0.0 [0.0-0.0]
0.0 [0.0 -70.0]
0.0 10.0-0.0]
0.0 [0.0-0.0]
0.0+++
[0.0-0.0]
41.5*
[9-92]
61.5
[49- 82]
12.5**+++
[9-60]
0.0 [0.0-0.0]
0.0 [0.0-0.0]
0.0 [0.0-0.0]
***; (P < 0.001), **; (P < 0.01). *; (P < 0.05) in comparison to Conlrol Gp.
+++; (P < 0.00 I). comparing Perindopril Pretreatment Stress Gp. (P-P S-Gp) to relevant stress Gps.
Heat and Light Stress: (H&L-S) I Cold Stress: (C-S) I Immobilization & Immersion Stress: (I&I-S).
Hormona.i Changes; Demonstrated in Table 3 and Figure (3)
Serum ACTH and cortisol, were significantly increased and testosterone was sig- nificantly decreased by H&L-S, in comparison to control. This profile was controlled by P-P H&L-S except for cortisol that persisted elevated in comparison to control. I&I-S significantly decreased testosterone arid increased the rest of hormones in comparison to control. p-p' I&I-S tended to confe~ little protection; as this profile, was pertained but at a lesser degree of significance except for PRL that reverted back to control. Thus PRL
!lnd cortisol were found significan.tly decreased by this regimen, when compared to I&I-S.
The Ability of Perindopril to..
35
Fig. I. Behavioural conduct changes in rats subjected to different modalities of stress with or without perindopril pretreatment.
Sec 700
200 100 0 Social Investigation Non-Sociallnvestigation
'Sec.
700 6(X) 500
400 300
3XJ 100 pop I&I-S \0
pop c-. ' pop C-S \ pop S-GpsI&I-S\
C-S \
~ :~;~:-
H& .
S-Gps.:co L-S
~ "(~!~~~
~Cortrol
Studied Gps Displacement Behavioural Conducts
***; (P < 0.001), **; (P < 0.01), *; (P < 0.05) in comparison to Control Gp -Perindopril Pretreatment Stress Gps (P-P- S- Gps)
Heat & Light Stress: (H&L-S) I Cold Stress: (C-S) I Immersion & Immobilization Stress: (I&I-S)
.ABLE
2. Changes in brain neurotransmitter concentrator of rats subjected to three different modalities of stress per se or after three weeks pretreatment with perindopril (400 ~g/kg) daily IP.
(Values are expressed as Mean:l: SE.) and compared using Student "t" test).
P- Pretreatment stress Gps Stress Gps
Studied Gps Control Gp I
C (0=6)
1&1 (0=6) H&L
(0=6)
C (0=6)
1&1 (n=6)
H&L (0=6) Brain
monamines (n=6)
1.3324+++
I
'1.2241*+++I
0.8281I 1,1720**to.1221 to,1853 to.OS31 to.1076
1.9657***
to.1481
0.8789"
t 0.092 NE in ug/g.
wet brain tissue
0.6521 to.0711
1.9794 :to.1218
2.5634"
:1:0.2412
1.7642 to.0928
2.6965"
t 0.3799
2.0921 t 0.5051
3.2589**
:t 0.3068
2.2114"
to.1718
DA in ug/g.
wet brain tissue
1.1213'1 1.326"
to.1763 to.1227
1.1463**
to.1302
0.9876"
:to,1198 0.6739
t 0,0573
0.8929 to.1291
0.9432 to.2193 SRr in Ug/g.
wet brain tissue
***; (P < 0.001). **; (P < 0.01), *; (P < 0.05) in comparison to Control Gp.
++; (P < 0.01), +++; (P < 0.001), when comparing Perindopril Pretreatment Stress Gps. (P-P S-Gps) to relevant Stress Gps.
Heat and Li2ht Stress; (H&L-S)! Cold Stress: (C-S) !Immersion & Immobilizatioo Stress: (I&I-S).
M.A. AI-Hazmi et al.
36
FIG. 2. Brain monamines' changes in rats subjected to different modalities of stress with or without per indopril pretreament.
!~
ugig Mtbrain tissue 35
- X..'. " " ",, . " 3
I ","..",
J
,'"~
" 225',' 1.5
-/I
.--"..
--
, 0ug/g Mt
bf8ln tiDue-- I
.35 r =
3~---
2.5 2 1.5
1 05
pop 1&1-5 ',:-
P-P Co.
pop C-5 '\---
p-p 5-Gps 1&1-5" ',~' ( ,
C.sH&L-5
X ~~
\""'""'""S-Gps CoItrol
~J~Qi~~ Gps
Serotonin Dopemine
Norepinephrine
Monamines
***; (P < 0.001). **; (P < 0.01), *; (P < 0.05) in comparison to Control Gp -Perindopril Pretreatment Stress Gps (P-P S-Gps I Heat & Light ~tress: (H&L-S) I Cold Stress: (C-S) I Immersion & Immobilization Stress: (I&I-S).
TABLE 3. Hormonal changes in rats subjected to three different modalities of stress per se or after three weeks pretreatment with perindopril (400 l1g/kg) daily JP.
[Values are expressed as Mean t SE.) and compared by using Student "(" test].
***; (P < 0.001), **; (P < 0.01), *; (P < 0.05) in comparison to Control Gp.
+; (P < 0.05), when comparing Perindopnl Pretreatment Stress Gps. (P-P S-Gps) to relevant Stress Gp&.
Heat and Light Stre&&; (H&L-S)! Cold Stress; (C-S) !Immersion & Immobilization Stress; (1%1-S).
The Abilily of Perindopril 10 37
***; (P < O.OOIJ, *", (P < 0.01, *;-(P < 0.05) in comparison to Control Gp. -Perindopril Pretreatment Stress Gps (P-P S-Gps) Heat & Light Stress: (H&L-S) I Cold Stress: (C-SJ I Immersion & Immobilization Stress: (I&I-SJ.
Histopathological Changes; Demonstrated in Plate-J
It illustrates that no structural testicular alterations were detected neither by S-Gps nor relevant P-P to S-Gps, in comparison to control.
Discussion
Justifying the validity of the concept that argues the "homogenicity of stressors" ver- sus the "homogenicity of responders" was one of the tasks wonhwhile probed. This is to panition whether &11 stressors evoke similar internal consequences that function the maximize survival in face of external predatory threats[211 or whether each has a par:' ticular profile of its own[41.
Answers to this have always been questionable, because difficulties always exist to standardized within the lab insults presenting in daily life. However, just by simultane- ously correlating the behavioural and neuroendocrine response outcome to the variable shon term modalitl"es of stress adopted in this study, an answer could be conspicuously speculated. Thus responses recorded appeared differential, i.e. stressor specific[21,221.
This is an issue now reponed in many studies clearing that responses as a whole are both adaptive in nature and patterned to environmental demand[4.22-241.
For instance, when present data are~viewed collectively, 17I-S appeared the most of- fending while C-S was the least. 1&1 insult comprises a complex of psychologically in-
38 M.A. AI-Huzmi trt ul.
teracting, componenrprovoked by immobilization and boostered by fear due to the dis- tressing existential element of fright from drowning[4]. Such stressful stimuli are abet to evoke immense behavioural and neuroendocrinal alterations(2,22] as currently dem- onstrated. The response pattern was different and more tamed by H&L-S probably as the psychological component "it. harbours was less distressing [being basically anxiety pr~voked by disruption of light-dark cycle[9.25J and is admixed ",itha physical com- ponent (heat exposure). The least response outcome was presently evoked by the phys- ical stimuli of cold exposure; whereby only brain NE content was increased and threat
,.
attempt suppressed in comparison to control.
Focusing on-each modality, the suppression of threat by C-S could De viewed as part of the patterning the environmental demand whereby all body resources were' reported
39
The Ability lJf PerindlJpriltlJ.
to be consumed in enhancing thermal metabolic reactions to restore basal metabolic rate to prestressor level, rather than being consumed in heightening any aggressive attemptl22,26,27].
Meanwhile the present demonstrable increase in NE contents; though appears con- troversial in literature, yet could still be explained secondary to an increase in cat- echolamine sensitizing sysiem triggered by cold exposure. This being mediated via a Ca calmod~lin dependent pathway, that was found to increase DA,[28] and may be legible to increase NE, just as presently observedo The absence of significant changes in 5HT by C-S were in line with reports probing its effect on different serotonin receptor sub- sets[29]0 The lack of other changes induced by cold exposure was also in accordance with findings in different literatures even through the exposure time was variable in such comparative studies{27.29,30].
Shifting to H&L stress, the non social conducts were increased, reflecting avoidance [the common denominator of anxiety in rodents][25] that prevailed at the expense of displacement. Usually different elements of displacement such as grooming are abet to predominate in restful states following stress, either as a rebound phenomenon or as a way to reduce arousal and redress imbalance caused by stressors[31,32]. However, this was not demonstrable in the present work, although most conditions necessary for its prevalence were existing. For example, increased displaceII)ent has been linked to in- creased ACTH; as currently observed and more so, for its parent CRH, as has been re- ported[33,34]. This is believed to be mediated via monaminergic transmission particular-
ly dopaminergic through stimulation of D2 receptor subsets,[35.36] and may even be through adrenergic or serotonergic[34,36,37],
Currently speaking, the brain content of the former two transmitters was found pres- ently elevated, and the increase in their activities were also reported to be associated with anxiety[38,39] while 5HT content in the current work was significantly altered which could be viewed to coincide with the anxiogenic predominance in this stress mo- dality studied and as confirmed in other reports emphasizing a role for serotonergic ag-
o d " k O hOb" 0
1
0 d od [4041]
omsts an serotomn reupta e In I Itors as anxlo ytlcs an anti epressants ' 0
The increase in environmental temperature encountered in H&L-S could have been an added factor to booster avoidance displacemento But this is rather questionable, as in- creased temperature was reported to markedly enhanced the activity of serotonergic neurons[7.41] and to induce generalized responses to ACTH;[42] to profile that was not totally fulfilled in the current study and that is debated by recent reports arguing a pos- sible role for serotonin in thermo-regulation[37]. Furthermore, it is relevant to add, that sleep depravation as a component of the H&L modality studied was reported per se by some not to playa major role in eliciting any neuroendocrine response[43]0
The concomitant suppression in testetorone observed whether by H&L or by I&I-S
..
could be appraised as part of the overall role of psychological stress on male re- production[44]o The main brunt of suppression is likely induced by the simultaneous el- evation in cortisol, where their reciprocal interrelationship has been coined[44,45]0 These reports, have cleared that the binding of cortisol to its receptors on Leydig cells sup- presses testosterone synthesis on testicular level[45], which can explain the current find-
40 M.A. A/-Haw!; et a/.
ings. While the possible involvement of LH; the controller of testosterone production, as a precipitating factor of suppression on pituitary level has been controversial in other studies;
confirmed by some[46] and denied by others[44], unfortunately not assessed in this study.
In H&L-S, elevated temperature could have been an additive factor to suppress tes- tosterone, but this was not presently observed as it was shown to be linked to increased PRL[42] that was not found elevated in this modality studied. In line, some reports claimed that PRL does not change in psychological stress while others emphasized on its increase by immobilization and restrain[44,47-49]. This was true in .the current 1&1 se- ries where increased PRL could have been an additive factor behind the more sig- nificant suppression in. testosterone observed in this modality. This increase was thought to involve tubuloinfundibuiar dopaminergic and serotonergic pathways[44,SO,SI]; which seems consistent to the finding that brain contents of both transmitters were found pres- ently e]evated by I&I-S. Moreover, immobilization per se was also reported to disrupt testicular steroidogenesis by inducing a state of Leydig cell hyposensitivity to gon- adotrophins[44,S2]; which leaves open a question that is not as yet settled and warrant as- sessment. However, it is worthwhile to note that the encountered changes by stress were only functional in some modalities while the testicular structure was totally spared.
Still speaking of I&I-S, the component of fear that prevailed could be viewed as the trigger to the increase in all neurotransmitters presently assessed. In this domain, it was previously stated that exposure to threatening stimuli activates the monoamine.rgic sys- tems in CNS, though in some articles they linked fear more to serotonin[41,S3], while in other they debated any specific role of this transmitter in stress[37,40]. However, what is agreed upon is that when animals are confronted by dangerous situations the emotional defensive behaviour predornin.ates and such offensive defensive pattern is dictated by the strength of the stressor[6,40,S3]. This was vividly observed in this study by the ~x- istential element of I&I-S that reset attempt to become withdrawn in defense at the ex- pense gf threat and has allowed its hangover even after the insult was terminated. The tendency for displacement to prevail over other elements of conduct was too a sur- prising finding as grooming was reported in literature to be immediately inhibited by se- vere stressors[31,32]. However, the likely explanation to this could be attributed to fur moisturing that was reported to -stimulate this acrr31], to partially takeover.
,It is apparent from all fore stated that response profile was specific to each modality and its behavioural neuroendocrinal magnitudes were dictated by the character of the stressor employed. It represents a very complex process and it is this complexity that is the cause of the individual response to stress[4,22].
At this juncture, it is relevant to clear that if these stimulatory processes to stress are unrestricted, the behavioural, neuroendocrinal, circulatory ...etc. of positive feedback loop would threaten rather than foster homeostasis during stress[S4]. Cortisol increase was natures protection to create negative feedback control loop to encompass such re- sponse[54]. ~ut with nowadays increase in stress intensity, duration, frequency, pre- dictabi]ity, chronicity... etc.[6] an urge to halt its etiopathologic consequences by ex- ogenous tools, has become mandatory; perindopril being assessed in this study.
The Abiiif." of Perindoprii to, 41
There appears that, perindopril pretreatment has fined tuned behavioural deficit in at- tempts and has promoted beneficial elements of conducts to take over. This was vividly
apparent as social investigations were significantly enhanced by the drug irrespective of the stress modality probed, This presents an important protective issue as it follows a pattern consistent with compounds possessing anxiolytic potential[55J. This is because, in part, it correlates well with concomitant significant increase in brain serotonin con- tent regardless of changes pre-induced by respective stress modalities; where interlinks between serotonin modulation and anxiolytic action was previously documented[25J, Moreover, the taming effect of perindopril pretreatment over the observed increase in NE and DA that was induced by the psychological component of 1&1 and H&L stresses, is considered beneficial, as it partially turned off those defense reactions controlling their overshooting rather than shutting them down, in such modalities.
The effect of perindopril on NE could be logically anticipated, knowing that many re- ports had observed its release from different brain areas by AG W56,57J, but the situa- tion remains questionable with respect to DA and 5HT[58-60J, Speaking of DA, its activ- ity in nigro-striatal pathways was reported to decrease with the concomitant decrease in Ag II and/or increase in bradykinin, substance P, enkephalines or neurotensin in the brain as a result of ACE inhibition by captopril[61J. In another study AG antagonists de- creased DA content in amygdala and entrorhinal cortex; all of which seem consistent to the present findings; though still others have claimed that interaction of Ag and DA re- ceptors are of intricate nature; the yield would favour the enhancement of some re- ceptors and block of others, dictating the same response[58,62J,
Shifting to serotonin the picture appeared more bizarre, as Ag antagonists were re- ported to increase serotonin in amygdala and striatum[63J while on the contrary Ag II was the one found to increase it in hypothalamus, brain stem and pineal gland[64], In a third study, Ag II reached on the rate limiting enzyme (tryptophane hydroxylase) re- sponsible for serotonin synthesis where it suppressed its synthesis at low Ag concentra- tions and enhanced it with higher levels[65J, which leaves open an area necessitating
through future probing. .
Nevertheless, it must be remembered that ACE in the brain does not apply hydrolyses Ag, but does so for other neuropeptides that dictate many behavioural and neuro- endocrinal outcomes elicited[65,66J. No wonder ACE have come under scrutiny and their ability to improve quality of life, to elevate mood and to improve cognitive functions has been coined[58,63,67,68] , Moreover, they possess predictive anxiolytic potentials based on findings relating Ag II to anxiety that is thought to be mediated through ATl receptor subs~ts in amygdala[58,64,67J, This is being further confirmed by findings that cleared the ability of Ag antagonist to reverse the behavioural deficits induced by stress
, , f
"
f " ,. ' bl d' [5869]and to potentIate the antIdepressant eJect 0 Imipramme m varIa e stu Ies ' . Another point of interest was the current observation that perindopril pretreatment succeeded to reset pituitary response to 1&1 and H&L-S at a lower pace. Meanwhile, it partially ameliorated the suppression on testosterone but could not sufficiently curtail cortisol pooling during the fore stated stress modalities, It is now established in literature
M.A. A/-HaZi
that endogenous brain Ag II can modulate formation and release of hypothalamic re- leasing factors[62] and that the peptide can also be released from the median eminence directly through the portal circulation[63,70]. Within the pituitary, Ag II has been ]ocal- ized in gonadotrophes of rats where they were reported in various publications to stim- ulate the paracrine release of PRL and ACTH via ATI receptors on lactotrophes and corticotrophes[62,71] and were also reported to exist by itself along with its synthesizing enzyme in human lactotrophes in PRL adenomas[72].
Beyond this, circulating Ag II was reported to act directly on the pituitary or via stim- ulating other circum ventricular organs, i.e. its acts on areas outside the blood brain bar- rier (BBB)[62]. Still to hold, the existence of anatomical links between Erg receptors and pathways inside the BBB and outside it indicates a close correlation between peripheral and central age system in the brain, this contributes to the regulation of hormonal, cir- culatory and fluid homeoastasis and the control of behaviour[62,64,70], as obseryed in varied literatures that were in line with some of perindopril actions observed in this study. Interesting enough, all raised areas outside the BBB were reported to be targets for ACE inhibitors administered systematically and recent reports have even suggested that this group of drugs to penetrate the brain[64] but the extent of penetration was argued to differ among individual inhibitors[64,73,74].
Nevertheless.. all forestated data when contrasted would lend credescence to the pos- sible mechanisms whereby perindopril could have impinged over to control PRL and ACTH overshooting when prophylactically given to modulate stress-~s observed" in this study. This fosters the raised question as to whether or not Ag neurons are essential components of the neuronal pathways that mediate ACTH increase and enhance the broad spectrum conditioner roles of PRL during stress[62,75]. This has been raised as the role of Ag in stress has been a subject of intense discussion and controversies[62,76]. The utility of. ACE is as tools to combat stress remains an open question that has been ad- dressed by perindopril findings in the present work.
The current improvement in testosterone profile encountered by perindopril pre- treatment to stress modalities studied is of special interest. This is because it partially coincides with perindopril modulation to pituitary secretion as far as PRL and ACm were concerned but remain unclear with respect to LH that was not currently assessed here, nor was its relation to brain and pituitary Ag totally settled in other reports[63,77].
The present study observed lack of appropriate suppression in cortisol which appeared partially dissociated from the concomitant changes in [ACTH] induced by perindopril pretreatment is not clear and needs an explanation. But fortunately, this seemed not to exert negative impacts on testicular steroidogenesis as it did not concomitantly induce a parallel suppression in the testosterone currently assessed. Also the likely possibility that increased brain serotonin content observed by perindopril pretreatment would in- crease PRL and subsidiary suppress testosterone[44,51], was luckily not recorded in the stress series presently probed. These protective issues need further justification although it must be admitted that most literature lack records on adverse effects of ACE-Is on tes- ticular functions which fits with the functional and structural findings linked to per- indopril pretreatment in the stressed rats currently studied. This could be more con-
The Ability of Perindoprirro.
43
firmed by salient reports that assessed the limited penetration of perindopril through the blood-testicular barrieJ78J that was potentiated by the lack of drug to modulate ACE intesticular semineferous tubules[79J; a finding that was also linked to quinalapril[801.
This study thus precluded that perindopril could be a useful tool to curtail many of the hazardous impacts of stress on psyche and soma by conferring anxiolytic inputs, meanwhile preserving male fertility profile. Further experimental work tempting to un- ravel more solid links on selective brain areas awaits to be assessed ahead before re- producing such advance in clinical trials then in practice.
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47
The Abiliry' of Perindopril fo
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