DOCA-salt hypertension and the role of the OVLT-sympathetic-gut microbiome axis.

Hypertension is a multifaceted condition influenced by genetic and environmental factors and estimated to cause 9.4 million deaths globally every year. Recently, there has been growing interest in understanding the gut microbe-host interaction in the maintenance of health or disease states, but relatively few studies have shown an association between the gut microbiome and specific types of hypertension. The deoxycorticosterone acetate (DOCA)-salt model of hypertension in rats is known to have a neurogenic component linked to increased sympathetic nervous system activity. As such, our lab has recently shown the hypertensive response in DOCA treated rats requires an intact organum vasculosum of the lamina terminalis (OVLT), a central hypothalamic circumventricular organ. Currently, we hypothesize the OVLT mediates changes in the gut microbiome associated with concomitant hypertension. Herein, we report that the hypertensive effects of DOCA-salt treatment were significantly attenuated throughout the 24-hour day/night cycle in OLVT lesioned rats on days 1, 3, and 9-21 of DOCA treatment compared with sham rats. Increased blood pressure (BP) in DOCA-salt treated rats was accompanied by specific changes in regional gut microbial populations yet was mitigated and offset by lesion of the OVLT. Furthermore, bacterial populations in OVLT-lesioned rats with attenuated hypertension more closely resembled those in normal control rats. We conclude that DOCA-salt hypertension is associated with specific microbiome changes in the gut, and the attenuated hypertensive effects of DOCA-salt in OVLT-lesioned rats is mediated in part through counteracting changes in these bacterial populations.

Development of the Deoxycorticosterone Acetate (DOCA)-salt Hypertensive Rat Model.

According to the World Health Organization (WHO), nearly 1.13 billion people worldwide have hypertension, a major factor responsible for premature death globally. The inherent multifactorial nature of hypertension makes its study difficult since the chronic rise in blood pressure depends on the intricate connection between dietary, genetic and environmental factors. Therefore, the pathophysi-ology of hypertension is not completely understood. For these reasons, there is an ongoing search for animal models that better mimic changes resulting from this disease. Because of its complexity, the use of animal models aimed at elucidating the pathogenesis of hypertension and to evaluate new therapeutic possibilities is an important tool for understanding this disease since it enables consistent experimental strategies that are impractical in humans. Over time, many animal models have been developed for the study of chronic increases in blood pressure ranging from genetic models that include the spontaneously hypertensive rat (SHR) and genetic manipulations, such as the TGR (mRen2) rat, as well as neurogenic or endocrine models. One of the most commonly used hypertensive rat models today is that of hypertension induced by treatment with deoxycorticosterone acetate associated with high sodium intake, i.e., the DOCA-salt model. This model is known to have a neurogenic component linked to increased sympathetic nervous system activity, and as such the DOCA-salt model promotes cross-talk between endocrine and neural components that lead to increased blood pressure, and may impact the functioning of other organs.

Lesion of the OVLT markedly attenuates chronic DOCA-salt hypertension in rats.

Lesions of the anteroventral third ventricle (AV3V region) are known to prevent many forms of experimental hypertension, including mineralocorticoid [deoxycorticosterone acetate (DOCA)-salt] hypertension in the rat. However, AV3V lesions include the organum vasculosum of the lamina terminalis (OVLT), portions of the median preoptic nucleus, and efferent fibers from the subfornical organ (SFO), thereby limiting the ability to define the individual contribution of these structures to the prevention of experimental hypertension. Having previously reported that the SFO does not play a significant role in the development of DOCA-salt hypertension, the present study was designed to test the hypothesis that the OVLT is necessary for DOCA-salt hypertension in the rat. In uninephrectomized OVLT-lesioned (OVLTx; n = 6) and sham-operated ( n = 4) Sprague-Dawley rats consuming a 0.1% NaCl diet and 0.9% NaCl drinking solution, 24-h mean arterial pressure (MAP) was recorded telemetrically 5 days before and 21 days after DOCA implantation (100 mg sc per rat). No differences in control MAP were observed between groups. The chronic pressor response to DOCA was attenuated in OVLTx rats such that MAP increased to 133 ± 3 mmHg in sham-operated rats by day 21 of DOCA compared with 120 ± 4 mmHg (means ± SE) in OVLTx rats. These results support the hypothesis that the OVLT is an important brain site of action for the pathogenesis of DOCA-salt hypertension in the rat.

Roles of the subfornical organ and area postrema in arterial pressure increases induced by 48-h water deprivation in normal rats.

In rats, water deprivation (WD) increases arterial blood pressure (BP) in part due to actions of elevated osmolality in the brain to increase vasopressin levels and sympathetic activity. However, the osmoreceptors that mediate this response have not been identified. To test the hypothesis that osmoregulatory circumventricular organs are involved, BP and heart rate (HR) were continuously recorded telemetrically during 48 h of WD in normal rats with lesions (x) or sham lesions (sham) of the subfornical organ (SFO) or area postrema (AP). Although WD increased BP in SFOx and SFOsham rats, no significant difference in the hypertensive response was observed between groups. HR decreased transiently but similarly in SFOx and SFOsham rats during the first 24 h of WD. When water was reintroduced, BP and HR decreased rapidly and similarly in both groups. BP (during lights off) and HR were both lower in APx rats before WD compared to APsham. WD increased BP less in APx rats, and the transient bradycardia was eliminated. Upon reintroduction of drinking water, smaller falls in both BP and HR were observed in APx rats compared to APsham rats. WD increased plasma osmolality and vasopressin levels similarly in APx and APsham rats, and acute blockade of systemic V1 vasopressin receptors elicited similar depressor responses, suggesting that the attenuated BP response is not due to smaller increases in vasopressin or osmolality. In conclusion, the AP, but not the SFO, is required for the maximal hypertensive effect induced by WD in rats.

OVLT lesion decreases basal arterial pressure and the chronic hypertensive response to AngII in rats on a high-salt diet.

We have reported that lesion of the organum vasculosum of the lamina terminalis (OVLT) has no effect on basal levels of mean arterial pressure (MAP) but abolishes the hypertensive effects of angiotensin II (AngII) in rats consuming a normal-salt diet. These results suggest that the OVLT does not contribute to regulation of MAP under conditions of normal salt intake, but it is an important brain site for the hypertensive actions of AngII. The OVLT has been proposed as a major sodium sensor in the brain and the hypertensive effects of AngII are exacerbated by high-salt intake. Therefore, the objective of this study was to investigate the role of the OVLT during AngII-induced hypertension in rats fed a high-salt diet. Male Sprague-Dawley rats underwent sham (Sham; n = 9) or OVLT lesion (OVLTx; n = 8) surgery and were placed on a high-salt (2% NaCl) diet. MAP was measured by radio telemetry during three control days, 10 days of AngII infusion (10 ng/kg/min, i.v.), and a 3-day recovery period. MAP was significantly lower in OVLTx (97 ± 2 mmHg) compared to Sham (106 ± 1 mmHg) rats during the control period (P < 0.05). Moreover, the chronic pressor response to AngII was markedly attenuated in OVLTx rats. MAP increased 58 ± 3 mmHg in Sham rats by Day 10 of AngII compared to a 40 ± 7 mmHg increase in OVLTx rats (P < 0.05). We conclude that (1) the OVLT regulates the basal levels of MAP in rats consuming a high-salt and (2) the OVLT is an important brain site of action for the pathogenesis of AngII-salt hypertension in the rat. Supported by HL076312.

Role of the organum vasculosum of the lamina terminalis for the chronic cardiovascular effects produced by endogenous and exogenous ANG II in conscious rats.

Endogenous and exogenous circulating ANG II acts at one of the central circumventricular organs (CVOs), the subfornical organ (SFO), to modulate chronic blood pressure regulation. However, at the forebrain, another important CVO is the organum vasculosum of the lamina terminalis (OVLT). In the present study, we tested the hypothesis that the OVLT mediates the hypertension or the hypotension produced by chronic infusion of ANG II or losartan (AT1 antagonist), respectively. Six days after sham or OVLT electrolytic lesion, male Sprague-Dawley rats (280-320 g, n = 6 per group) were instrumented with intravenous catheters and radiotelemetric blood pressure transducers. Following another week of recovery, rats were given 3 days of saline control infusion (7 ml/day) and were then infused with ANG II (10 ng·kg(-1)·min(-1)) or losartan (10 mg·kg(-1)·day(-1)) for 10 days, followed by 3 recovery days. Twenty-four hour average measurements of mean arterial pressure (MAP) and heart rate (HR) were made during this protocol. Hydromineral balance (HB) responses were measured during the experimental protocol. By day 9 of ANG II treatment, MAP had increased 16 ± 4 mmHg in sham rats but only 4 ± 1 mmHg in OVLT lesioned rats without changes in HR or HB. However, the hypotension produced by 10 days of losartan infusion was not modified in OVLT lesioned rats. These results suggest that the OVLT might play an important role during elevation of plasma ANG II, facilitating increases of blood pressure but is not involved with baseline effects of endogenous ANG II.

Simultaneous administration of Ang(1-7) or A-779 does not affect the chronic hypertensive effects of angiotensin II in normal rats.

HYPOTHESIS: The following studies were designed to test the hypothesis that simultaneous administration of either Ang(1-7) or its antagonist A-779 would affect the chronic hypertensive effects of angiotensin II (Ang II). INTRODUCTION: Despite the well-described actions of Ang(1-7) and its role possessing opposite actions to Ang II, there have been few studies examining the role of Ang(1-7) in a chronic setting. It is well established that Ang(1-7) plays a protective role in preventing deleterious effects of Ang II in the heart, but little is known of its role in modulating the chronic hypertensive effects of Ang II. MATERIALS AND METHODS: Rats were instrumented with venous catheters and telemetric pressure transducers. Arterial pressure responses were measured in rats treated with Ang II (10 ng/kg/ min) (n=9) and compared with those treated with Ang II and Ang(1-7) (24 microg/kg/h) (n=8), or the Ang(1-7) antagonist A-779 (24 microg/kg/h) (n=7) for 8 days. RESULTS: Mean arterial pressure rose significantly and similarly in all three groups of rats, such that by day 8 of Ang II infusion, pressures had risen 25-30 mmHg in all rats. CONCLUSIONS: These results do not support the hypothesis that the chronic hypertensive effects of Ang II in normal rats are altered by co-administration of either Ang(1-7) or A-779.

The cardiovascular response of normal rats to dual lesion of the subfornical organ and area postrema at rest and to chronic losartan.

The subfornical organ (SFO) and the area postrema (AP), two of the sensory circumventricular organs (CVO), are known to play a role in the chronic central control of blood pressure. In previous studies in which these regions were independently lesioned, the chronic hypotensive effects of the AT(1) receptor blocker losartan (10 mg/kg/day) were attenuated by ~15 mm Hg. In the present study, we sought to investigate the effect of concurrent lesion of both the SFO and the AP on the cardiovascular effects of chronic losartan infusion in order to test the hypothesis that a greater attenuation of the hypotensive effects of losartan would be observed in rats with dual lesions. To do so, arterial pressure and heart rate responses to 10-day infusion of losartan were compared in sham rats and those with dual lesions of the AP and SFO. Two important findings resulted from this study. First, dual lesion rats exhibited a sustained and significant decrease in resting blood pressure (83+/-1 mm Hg vs. 104+/-1 mm Hg, respectively) and heart rate (356+/-3 bpm vs. 398+/-6 bpm, respectively) compared to sham animals. Secondly, rats with concurrent lesion of both the AP and the SFO demonstrated a significantly attenuated response to losartan compared to sham animals but showed no greater attenuation of losartan's chronic hypotensive effects than animals with lesion of either the SFO or the AP (approximately 15 mm Hg). Although these results do not support the stated hypothesis, they do suggest redundancy and compensatory roles of the AP and SFO in basal cardiovascular control.

Changing dietary sodium alters the chronic cardiovascular effects of losartan in rats.

INTRODUCTION: We have previously demonstrated a profound hypotensive response to the angiotensin II type 1 (AT1)-receptor antagonist losartan in rats consuming a normal salt diet that is not seen in salt-loaded rats, presumably due to a suppression of the renin-angiotensin system (RAS) by high sodium levels. The purpose of the present study was to examine the cardiovascular effects of changing dietary sodium intake during chronic treatment with losartan. We hypothesised that during blockade of AT1-receptors by chronic losartan infusion, when renin levels would be elevated regardless of dietary sodium, changing diets from high to normal or normal to high salt would have no effect on mean arterial pressure (MAP). MATERIALS AND METHODS: To test this hypothesis, groups of rats instrumented with radiotelemetry transducers for MAP monitoring and venous catheters for infusion were initially placed on either a 0.4% salt content diet, referred to as Losartan Normal diet - High salt diet (LosN-HI, n = 7), or a 4.0% salt content diet, referred to as Losartan High salt diet - Normal diet (LosHI-N, n = 9). After a thee-day control period, infusion of losartan was begun in all rats (10 mg/kg/day in 7 ml/day isotonic saline i.v.). After 10 days, diets were switched between groups and data were collected for another 10 days, after which losartan infusion was terminated for a 10-day recovery period. RESULTS: At the start of losartan infusion MAP was observed to be similar between LosN-HI rats (101+/-2 mmHg) and LosHI-N rats (101+/-2 mmHg). By day seven of the first 10 day protocol, MAP in LosN-HI rats had fallen to 71+/-4 mmHg while decreasing to 90+/-2 mmHg in LosHI-N rats. Five days after switching diets, MAP in LosN-HI rats had risen back to 85+/-3 mmHg, while MAP in LosHI-N rats had fallen to 75+/-2 mmHg. CONCLUSIONS: These results do not support our hypothesis, suggesting that changing dietary sodium can alter the chronic hypotensive response to losartan regardless of the initial state of the RAS.

ANG II-induced hypertension and the role of the area postrema during normal and increased dietary salt.

It has been shown that the area postrema (AP) plays a role in the development of certain types of chronic angiotensin II (ANG II)-induced hypertension in the rat but is not of great importance in the salt sensitivity of arterial pressure. It has recently been proposed, however, that elevated sodium levels may exacerbate the hypertensive effects of ANG II, which by itself dramatically affects salt sensitivity, by acting at sodium-sensing neurons in certain circumventricular organs of the brain. Thus the interactions of ANG II, sodium, and the central nervous system remain to be fully understood. The purpose of this study was to examine the role of the AP in ANG II-induced hypertension during periods of normal and elevated dietary salt. We hypothesized that an intact AP was necessary for the full development of hypertension under chronic ANG II infusion and that its role would be pronounced during periods of increased dietary sodium. To test this, male Sprague-Dawley rats underwent ablation of the area postrema (APx, n = 6) or sham operation (sham, n = 6). After 3 wk of recovery, rats were instrumented with radiotelemetry transducers for constant blood pressure and heart rate monitoring and venous catheters for vehicle infusion. After a 3-day control period of 0.9% saline infusion (7 ml/day) and 0.4% dietary sodium, a 10-day period of ANG II infusion (10 ng.kg(-1).min(-1)) was begun, immediately followed by a second 10-day period during which rats were fed a 4.0% sodium diet. By day 6 of ANG II infusion, mean arterial pressure (MAP) in APx rats had increased to 139 +/- 4 mmHg, whereas MAP in sham rats had increased to 126 +/- 3 mmHg. This difference was found to be significant and continued through day 1 of the high-salt period, after which MAP of the two groups had risen to similar levels. On day 9 of high salt, MAP was again observed to be significantly higher (162 +/- 1 mmHg) in APx rats when compared with sham rats (147 +/- 4 mmHg.) These results do not support the hypothesis that the AP is necessary for the full development of ANG II-induced hypertension at normal or elevated levels of dietary sodium.