|Year : 2016 | Volume
| Issue : 2 | Page : 93-97
Orchidectomy reduces blood pressure, but testosterone increases it in intact and orchidectomized normotensive rats
Fati O Aliyu1, Abdullateef I Alagbonsi2, Ali AU Dikko3
1 Department of Human Physiology, Faculty of Medicine, Ahmadu Bello University, Zaria, Kaduna State, Nigeria
2 Department of Standards and Quality Assurance, National Health Insurance, Scheme, North Central A Zonal Office, Kwara State Ministry of Health Premises, Fate; Department of Physiology, College of Health Sciences, University of Ilorin, Ilorin, Kwara State, Nigeria
3 Department of Physiology, Faculty of Medicine, Bayero University, Kano, Kano State, Nigeria
|Date of Web Publication||4-Aug-2016|
Abdullateef I Alagbonsi
Department of Standards and Quality Assurance, National Health Insurance Scheme, North Central A Zonal Office, Kwara State Ministry of Health Premises, Fate, Ilorin, Kwara
Source of Support: None, Conflict of Interest: None
Background: The cardioprotective effect of estrogen has been well documented, and the effect of testosterone on blood pressure (BP) is controversial and remains inconclusive.
Aim: The present study is aimed at investigating the effect of testosterone on BP in normotensive rats.
Materials and Methods: In a blind study, 30 male albino rats (200-250 g) were divided into 5 oral treatment groups (n = 6 rats each) as follows: Groups I and II were intact rats that received 1 ml/kg normal saline (vehicle) and 25 mg/kg testosterone subcutaneously for 24 days. Group III was sham-operated and received normal saline for 24 days. Groups IV and V were bilaterally orchidectomized and received normal saline and 25 mg/kg testosterone subcutaneously for 24 days.
Results: The systolic BP (SBP), diastolic BP (DBP), pulse pressure (PP), and mean arterial pressure (MAP) of the sham-operated rats that received normal saline were not significantly different from the intact (control) rats that received normal saline (P > 0.05). Testosterone treatment in control rats caused a sustainably higher SBP (P < 0.001), DBP (P < 0.001), PP (P < 0.001), and MAP (P < 0.001) when compared to control that received normal saline. The SBP (P < 0.01), DBP (P < 0.001), MAP (P < 0.001), but not PP (P > 0.05) was significantly reduced in orchidectomized rats that received normal saline when compared to control. However, administration of testosterone in orchidectomized rats abolished orchidectomy-induced reduction in SBP, DBP, MAP, but not PP and further increased them above the control level (P < 0.05).
Conclusion: This study shows that testosterone is prohypertensive in normotensive rats.
Keywords: Blood pressure, hypertension, orchidectomy, testosterone
|How to cite this article:|
Aliyu FO, Alagbonsi AI, Dikko AA. Orchidectomy reduces blood pressure, but testosterone increases it in intact and orchidectomized normotensive rats. Nig J Cardiol 2016;13:93-7
|How to cite this URL:|
Aliyu FO, Alagbonsi AI, Dikko AA. Orchidectomy reduces blood pressure, but testosterone increases it in intact and orchidectomized normotensive rats. Nig J Cardiol [serial online] 2016 [cited 2019 Sep 19];13:93-7. Available from: http://www.nigjcardiol.org/text.asp?2016/13/2/93/187705
| Introduction|| |
Generally, males are at greater risk for cardiovascular and renal diseases than their age-matched premenopausal females. This created an impression that gender influences blood pressure (BP) and sex hormones contribute to the pathophysiology of hypertension.  In humans, for instance, 24-h ambulatory BP is higher in men than in women of similar ages. ,,, In different hypertensive rat models, many investigators have found that males have higher BP than do females. ,,,,,,,, However, there was evidence of higher BP in postmenopausal women than their age-matched men. 
The factor(s) responsible and the mechanism(s) involved in the increased BP in male rats and perhaps in elderly female rats remain to be fully understood, but androgens have been implicated.  For instance, data from some studies clearly showed that BP is higher in boys than in girls during adolescence and puberty, when androgen levels are increasing. , Similarly, castration at a young age (3-5 weeks) was reported to attenuate the development of hypertension in different hypertensive models. ,,,,,,,,, Furthermore, chronic blockade of androgen receptor with the antagonist flutamide attenuates BP in male spontaneously hypertensive rats (SHR) to the level found in female SHR.  Perhaps the observed gender differences and the increased incidence of cardiac events in women after menopause are not entirely due to the depletion of estrogen but are also related to significant amounts of circulating testosterone, because postmenopausal ovaries reportedly produce significant amounts of androgens in the form of testosterone and androstenedione. 
However, there are other suggestions that female sex hormones may actually protect against a salt-induced increase in BP, possibly by augmenting the renal excretion of sodium.  Thus, when Dahl salt-sensitive (DS) rats receive a high-sodium diet, female rats become less hypertensive than male rats.  In this animal model, gonadectomy results in an accelerated development of salt-sensitive hypertension in females. The increase in salt sensitivity observed after ovariectomy is actually associated with a blunted pressure-natriuresis relationship. Interestingly, reversal of the diet to a low salt intake reverses the hypertension in intact male and female DS rats, but this was not the case in ovariectomized female DS rats, suggesting that female sex hormones act to suppress sodium-dependent as well as sodium-independent increases in BP.  A greater rise in BP also occurred in spontaneously hypertensive female rats after ovariectomy. ,
Testosterone replacement therapy is now being prescribed more often for aging men, the same population in which BP and prostate cancer incidence increases. The hypothesis that high levels of circulating androgens are a risk factor for prostate cancer is supported by the dramatic regression of the cancer after castration.  Over recent decades, testosterone has been increasingly abused for muscle buildings and enhancement of physical performance,  and the side effects on the cardiovascular system may include elevated BP  and harmful cholesterol levels leading to increased risk of cardiovascular diseases and coronary artery diseases.
The cardioprotective effect of estrogen has been extensively demonstrated. However, the hypothesis that androgens may contribute to cardiovascular risk is still debated. While some studies show that testosterone can attenuate BP through relaxation of the vascular smooth muscle,  other studies demonstrate its prohypertensive effect by the constriction of vascular smooth muscle.  Since majority of previous studies have established the prohypertensive effect of testosterone in hypertensive models, the present study is aimed at investigating the effect of testosterone on BP in normotensive rats.
| Materials and methods|| |
Thirty male albino rats (200-250 g) were housed at room temperature with free access to food and water ad libitum and were maintained on a 12-h light/dark cycle, with the lights on from 7:00 am "Principles of laboratory animal care (NIH publication No. 85-23, revised 1985)" were followed. All experiments have been examined and approved by our Institutional Ethics Committee.
After 2 weeks acclimatization to their new environment with standard laboratory diet and water given ad libitum, the 30 animals were randomly divided in a blinded fashion into 5 oral treatment groups as follows:
- Group I served as control and received 1 ml/kg normal saline (vehicle) for 24 days
- Group II received 25 mg/kg testosterone subcutaneously for 24 days
- Group III was sham-operated and received normal saline for 24 days
- Group IV was orchidectomized and received normal saline for 24 days
- Group V was orchidectomized and received 25 mg/kg testosterone subcutaneously for 24 days.
Each rat was anesthetized using chloroform, it was then placed on a dissecting board, and a pair of scissors was used to make a skin incision on the two scrotums. Bilateral orchidectomy was induced by removal of the two testes from the scrotums. Then chromic catgut suture was used to suture the incision, followed by a subcutaneous injection of 0.5 ml ampicillin to prevent infection. 
The procedure for sham operation was similar except that the testes were not removed from the scrotum.
Measurement of blood pressure
BP was measured by tail-cuff method of Buñag and Butterfield  as modified by Byrom and Wilson  using a noninvasive Ugo Basile, series 5b500 BP recorder. Each rat was placed in the restrainer and kept in a scanner for 30 min to warm the animal prior to obtaining pressure measurement. A cuff was placed on the base of the tail to occlude the blood flow. A transducer was placed close to the cuff which measured the systolic BP (SBP), diastolic BP (DBP), pulse pressure (PP), and mean arterial pressure (MAP). Upon deflation, the noninvasive BP sensor was used to monitor the BP, and an average of four readings was taken for each rat so as to ensure accuracy.
Data were analyzed with one-way ANOVA using GraphPad Prism (Version 188.8.131.52), followed by a post hoc Tukey test for multiple comparisons. Data were presented as the mean ± standard error of the mean. P ≤ 0.05 were considered statistically significant.
| Results|| |
The SBP, DBP, PP, and MAP of the sham-operated rats that received normal saline were not significantly different from the control rats that received normal saline (P > 0.05). Testosterone treatment in control rats caused a sustainably higher SBP (P < 0.001), DBP (P < 0.001), PP (P < 0.001), and MAP (P < 0.001) when compared to control that received normal saline. The SBP (P < 0.01), DBP (P < 0.001), MAP (P < 0.001), but not PP (P > 0.05) were significantly reduced in orchidectomized rats that received normal saline when compared to control. However, administration of testosterone in orchidectomized rats abolished orchidectomy-induced reduction in SBP, DBP, MAP, but not PP and further increased them above the control level (P < 0.05) [Figure 1] [Figure 2] [Figure 3] [Figure 4].
|Figure 1: Effect of testosterone on SBP in normal and orchidectomized rats. Values are expressed as mean ± standard error of the mean (n = 4). SBP - Systolic blood pressure|
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|Figure 2: Effect of testosterone on DBP in normal and orchidectomized rats. Values are expressed as mean ± standard error of the mean (n = 4). DBP - Diastolic blood pressure|
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|Figure 3: Effect of testosterone on PP in normal and orchidectomized rats. Values are expressed as mean ± standard error of the mean (n = 4). PP - Pulse pressure|
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|Figure 4: Effect of testosterone on MAP in normal and orchidectomized rats. Values are expressed as mean ± standard error of the mean (n = 4). MAP - Mean arterial pressure|
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| Discussion|| |
Gonadal hormones have been suggested to be responsible for the difference in BP found in males and females. However, previous studies on the effects of androgens and estrogens on BP are contradictory and remain inconclusive. , The effect of testosterone on BP in normotensive rats was investigated in this study.
In the present study, supplementation of normotensive rats with testosterone persistently caused an increase in BP of rats and consequently elicit hypertension (BP >140/90 mmHg). Previously, low prevalence of coronary disease among men with hypotestosteronemia plus hyperestrogenemia  and increased risk of coronary artery disease and myocardial infarction in women suffering from chronic anovulation and exhibiting hypertestosteronemia have been documented. On the contrary, lower or unchanged circulating testosterone in hypertensive men ,, and in men with coronary artery disease  or myocardial infarction  has also been reported. These studies suggest that decreased, rather than increased, androgen levels are associated with hypertension, myocardial infarction, and coronary artery disease. However, the cardioprotective property of testosterone speculated by these aforementioned studies have been criticized by other authors who attributed the low testosterone level in cardiovascular diseases to stress, since reduced testosterone level has been shown to be a consequence of stress elicited by myocardial infarction, surgery, head trauma, burns, hypoxia, sleep deprivation, and psychological stressors.  The present study, taken together with these previous reports, support the contention that testosterone is a prohypertensive hormone and contribute to the higher BP generally observed in men when compared to women of similar age.
Having observed the prohypertensive effect of testosterone supplement in normotensive rats, we went further to investigate if testosterone deficiency will sustainably reduce BP in normotensive rats. It was observed that orchidectomy-induced testosterone deficiency caused a persistent decrease in BP in normotensive rats and consequently elicit hypotension (BP <90/60 mmHg). This finding is similar to the previous observation that orchidectomy at a young age (3-5 weeks) attenuated the development of hypertension in spontaneously hypertensive and DS rats. ,,,,,
In order to conclusively demonstrate the unique contribution of testosterone to the development of high BP, we investigated whether testosterone replacement in orchidectomized rats can potently reverse the BP to the level similar to the control. In the present study, supplementation of orchidectomized rats with testosterone not only completely abolished the orchidectomy-induced hypotension, but elevated it slightly above the control level. This observation is similar to the previous study where testosterone replacement in orchidectomized rats increased BP to levels similar to those in intact rats. ,
Some previous studies have suggested mechanisms by which testosterone could elevate BP and damage blood vessels. For instance, testosterone was reported to increase circulating level of homocysteine, which induces endothelial damage, thus leading to the development of atherosclerosis, and may adversely influence renal function by damaging glomerular endothelial cells.  In addition, testosterone increases endothelin-1 levels in subjects undergoing a sex change, suggesting a way through which it may cause BP elevation. , Moreover, elevated catecholamine levels in SHR are associated with high BP, , whereas castration reduces it to those observed in normotensive control rats,  suggesting that testosterone can also induce catecholamine synthesis.  This observation could be mediated by the stimulation of tyrosine hydroxylase which is the rate-limiting enzyme for catecholamine synthesis. , Moreover, testosterone also stimulates angiotensin expression  and increases circulating levels of angiotensin-converting enzyme, thereby causing hypertension. 
| Conclusion|| |
The present study provides convincing evidence for the prohypertensive effect of testosterone in normotensive rats.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Ouchi Y, Share L, Crofton JT, Iitake K, Brooks DP. Sex difference in the development of deoxycorticosterone-salt hypertension in the rat. Hypertension 2009;9:172-7.
Burt VL, Whelton P, Roccella EJ, Brown C, Cutler JA, Higgins M, et al.
Prevalence of hypertension in the US adult population. Results from the third national health and nutrition examination survey, 1988-1991. Hypertension 1995;25:305-13.
Khoury S, Yarows SA, O'Brien TK, Sowers JR. Ambulatory blood pressure monitoring in a nonacademic setting. Effects of age and sex. Am J Hypertens 1992;5:616-23.
Staessen J, Fagard R, Lijnen P, Thijs L, van Hoof R, Amery A. Reference values for ambulatory blood pressure: A meta-analysis. J Hypertens Suppl 1990;8:S57-64.
Wiinberg N, Høegholm A, Christensen HR, Bang LE, Mikkelsen KL, Nielsen PE, et al.
24-h ambulatory blood pressure in 352 normal Danish subjects, related to age and gender. Am J Hypertens 1995;8 (10 Pt 1):978-86.
Ashton N, Balment RJ. Sexual dimorphism in renal function and hormonal status of New Zealand genetically hypertensive rats. Acta Endocrinol (Copenh) 1991;124:91-7.
Chen YF, Meng QC. Sexual dimorphism of blood pressure in spontaneously hypertensive rats is androgen dependent. Life Sci 1991;48:85-96.
Crofton JT, Ota M, Share L. Role of vasopressin, the renin-angiotensin system and sex in Dahl salt-sensitive hypertension. J Hypertens 1993;11:1031-8.
Crofton JT, Share L. Gonadal hormones modulate deoxycorticosterone-salt hypertension in male and female rats. Hypertension 1997;29(1 Pt 2):494-9.
Ganten U, Schröder G, Witt M, Zimmermann F, Ganten D, Stock G. Sexual dimorphism of blood pressure in spontaneously hypertensive rats: Effects of anti-androgen treatment. J Hypertens 1989;7:721-6.
Iams SG, McMurthy JP, Wexler BC. Aldosterone, deoxycorticosterone, corticosterone, and prolactin changes during the lifespan of chronically and spontaneously hypertensive rats. Endocrinology 1979;104:1357-63.
Masubuchi Y, Kumai T, Uematsu A, Komoriyama K, Hirai M. Gonadectomy-induced reduction of blood pressure in adult spontaneously hypertensive rats. Acta Endocrinol (Copenh) 1982;101:154-60.
Ouchi Y, Share L, Crofton JT, Iitake K, Brooks DP. Sex difference in the development of deoxycorticosterone-salt hypertension in the rat. Hypertension 1987;9:172-7.
Rowland NE, Fregly MJ. Role of gonadal hormones in hypertension in the Dahl salt-sensitive rat. Clin Exp Hypertens A 1992;14:367-75.
Staessen J, Fagard R, Lijnen P, Thijs L, van Hoof R, Amery A. Reference values for ambulatory blood pressure: A meta-analysis. J Hypertens 2010;8:S57-64.
Bachmann H, Horacek U, Leowsky M, Hirche H. Blood pressure in children and adolescents aged 4 to 18. Correlation of blood pressure values with age, sex, body height, body weight and skinfold thickness (Essen Blood Pressure Study). Monatsschr Kinderheilkd 1987;135:128-34.
Harshfield GA, Alpert BS, Pulliam DA, Somes GW, Wilson DK. Ambulatory blood pressure recordings in children and adolescents. Pediatrics 1994;94(2 Pt 1):180-4.
Gafter U, Ben-Bassat M, Levi J. Castration inhibits glomerular hypertrophy and proteinuria in uninephrectomized male rats. Eur J Clin Invest 1990;20:360-5.
Iams SG, Wexler BC. Retardation in the development of spontaneous hypertension in SH rats by gonadectomy. J Lab Clin Med 1977;90:997-1003.
Iams SG, Wexler BC. Retardation in the development of spontaneous hypertension in SH rats by gonadectomy. J Lab Clin Med 2007;90:997-1003.
Lombet JR, Adler SG, Anderson PS, Nast CC, Olsen DR, Glassock RJ. Sex vulnerability in the subtotal nephrectomy model of glomerulosclerosis in the rat. J Lab Clin Med 1989;114:66-74.
Mályusz M, Ehrens HJ, Wrigge P. Effect of castration on the experimental renal hypertension of the rat. Blood pressure, nephrosclerosis, long-chain fatty acids, and N-acetylation of PAH in the kidney. Nephron 1985;40:96-9.
Reckelhoff JF, Zhang H, Srivastava K, Granger JP. Gender differences in hypertension in spontaneously hypertensive rats: Role of androgens and androgen receptor. Hypertension 1999;34(4 Pt 2):920-3.
Sluijmer AV, Heineman MJ, De Jong FH, Evers JL. Endocrine activity of the postmenopausal ovary: The effects of pituitary down-regulation and oophorectomy. J Clin Endocrinol Metab 1995;80:2163-7.
Kensicki E, Dunphy G, Ely D. Estradiol increases salt intake in female normotensive and hypertensive rats. J Appl Physiol 2002;93:479-83.
Crofton JT, Ota M, Share L. Role of vasopressin, the renin-angiotensin system, and sex in Dahl salt-sensitive hypertension. J Hypertens 2013;11:1031-8.
Chen YF, Meng QM. Sexual dimorphism of blood pressure in spontaneously hypertensive rats is androgen dependent. Life Sci 2006;48:85-96.
Calhoun DA, Zhu ST, Chen YF, Oparil S. Gender and dietary NaCl in spontaneously hypertensive and Wistar-Kyoto rats. Hypertension 1995;26:285-9.
Ashton N, Balment RJ. Sexual dimorphism in renal function and hormonal status of New Zealand genetically hypertensive rats. Acta Endocrinol (Copenh) 2009;124:91-7.
Raynaud JP. Prostate cancer risk in testosterone-treated men. J Steroid Biochem Mol Biol 2006;102:261-6.
Handelsman DJ. Testosterone: Use, misuse and abuse. Med J Aust 2006;185:436-9.
Hartgens F, Kuipers H. Effects of androgenic-anabolic steroids in athletes. Sports Med 2004;34:513-54.
Cairrão E, Alvarez E, Santos-Silva AJ, Verde I. Potassium channels are involved in testosterone-induced vasorelaxation of human umbilical artery. Naunyn Schmiedebergs Arch Pharmacol 2008;376:375-83.
Herman SM, Robinson JT, McCredie RJ, Adams MR, Boyer MJ, Celermajer DS. Androgen deprivation is associated with enhanced endothelium-dependent dilatation in adult men. Arterioscler Thromb Vasc Biol 1997;17:2004-9.
Buñag RD, Butterfield J. Tail-cuff blood pressure measurement without external preheating in awake rats. Hypertension 1982;4:898-903.
Byrom FB, Wilson C. A plethysmographic method for measuring systolic blood pressure in the intact rat. J Physiol 2007;93:301-4.
Phillips GB, Pinkernell BH, Jing TY. The association of hypotestosteronemia with coronary artery disease in men. Arterioscler Thromb 1994;14:701-6.
Hughes GS, Mathur RS, Margolius HS. Sex steroid hormones are altered in essential hypertension. J Hypertens 1989;7:181-7.
Jaffe A, Chen Y, Kisch ES, Fischel B, Alon M, Stern N. Erectile dysfunction in hypertensive subjects. Assessment of potential determinants. Hypertension 1996;28:859-62.
Phillips GB, Jing TY, Resnick LM, Barbagallo M, Laragh JH, Sealey JE. Sex hormones and hemostatic risk factors for coronary heart disease in men with hypertension. J Hypertens 1993;11:699-702.
Kalin MF, Zumoff B. Sex hormones and coronary disease: A review of the clinical studies. Steroids 1990;55:330-52.
Reckelhoff JF, Zhang H, Granger JP. Testosterone exacerbates hypertension and reduces pressure-natriuresis in male spontaneously hypertensive rats. Hypertension 1998;31(1 Pt 2):435-9.
Reckelhoff JF, Zhang H, Srivastava K. Gender differences in development of hypertension in spontaneously hypertensive rats: Role of the renin-angiotensin system. Hypertension 2000;35(1 Pt 2):480-3.
Giltay EJ, Hoogeveen EK, Elbers JM, Gooren LJ, Asscheman H, Stehouwer CD. Effects of sex steroids on plasma total homocysteine levels: A study in transsexual males and females. J Clin Endocrinol Metab 1998;83:550-3.
Oloyo AK, Vineetha VP, Anigbogu CN, Shofola OA. Orchidectomy ameliorates the vascular hypertrophic effect of a high salt diet in Sprague-Dawley rats. J Afr Assoc Physiol Sci 2013;1:37-45.
Kumai T, Tanaka M, Watanabe M, Matsumoto C, Kobayashi S. Possible involvement of androgen in increased norepinephrine synthesis in blood vessels of spontaneously hypertensive rats. Jpn J Pharmacol 1994;66:439-44.
Kumai T, Tanaka M, Watanabe M, Nakura H, Kobayashi S. Influence of androgen on tyrosine hydroxylase mRNA in adrenal medulla of spontaneously hypertensive rats. Hypertension 1995;26:208-12.
Leung PS, Wong TP, Lam SY, Chan HC, Wong PY. Testicular hormonal regulation of the renin-angiotensin system in the rat epididymis. Life Sci 2000;66:1317-24.
Lim YK, Retnam L, Bhagavath B, Sethi SK, bin Ali A, Lim SK. Gonadal effects on plasma ACE activity in mice. Atherosclerosis 2002;160:311-6.
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