Stress In Spontaneously Hypertensive Rats Research Articles

MiR-196a-5p hinders vascular smooth muscle cell proliferation and vascular remodeling via repressing BACH1 expression

Hyperproliferation of vascular smooth muscle cells (VSMCs) is a driver of hypertensive vascular remodeling. This study aimed to uncover the mechanism of BTB and CNC homology 1 (BACH1) and microRNAs (miRNAs) in VSMC growth and hypertensive vascular remodeling. With the help of TargetScan, miRWalk, miRDB, and miRTarBase online database, we identified that BACH1 might be targeted by miR-196a-5p, and overexpressed in VSMCs and aortic tissues from spontaneously hypertensive rats (SHRs). Gain- and loss-of-function experiments demonstrated that miR-196a-5p suppressed VSMC proliferation, oxidative stress and hypertensive vascular remodeling. Double luciferase reporter gene assay and functional verification showed that miR-196a-5p cracked down the transcription and translation of BACH1 in both Wistar Kyoto rats (WKYs) and SHRs. Silencing BACH1 mimicked the actions of miR-196a-5p overexpression on attenuating the proliferation and oxidative damage of VSMCs derived from SHRs. Importantly, miR-196a-5p overexpression and BACH1 knockdown cooperatively inhibited VSMC proliferation and oxidative stress in SHRs. Furthermore, miR-196a-5p, if knocked down in SHRs, aggravated hypertension, upregulated BACH1 and promoted VSMC proliferation, all contributing to vascular remodeling. Taken together, targeting miR-196a-5p to downregulate BACH1 may be a promising strategy for retarding VSMC proliferation and hypertensive vascular remodeling.

Inhibition of endoplasmic reticulum stress restores the balance of renal RAS components and lowers blood pressure in the spontaneously hypertensive rats

ABSTRACT Background Endoplasmic reticulum (ER) stress has been shown to play a critical role in the pathogenesis of hypertension. However, the underlying mechanisms for lowering blood pressure (BP) by suppressing ER stress remain unclear. Here, we hypothesized that inhibition of ER stress could restore the balance between RAS components and lower BP in spontaneously hypertensive rats (SHRs). Methods Wistar-Kyoto (WKY) rats and SHRs received vehicle or 4-PBA, an ER stress inhibitor, in the drinking water for 4 weeks. BP was measured by tail-cuff plethysmography, and the expression of RAS components was examined by Western blot. Results Compared with vehicle-treated WKY rats, vehicle-treated SHRs exhibited higher blood pressure and increased renal ER stress and oxidative stress, accompanied by impaired diuresis and natriuresis. Moreover, SHRs had higher ACE and AT1R and lower AT2R, ACE2, and MasR expressions in the kidney. Interestingly, 4-PBA treatment improved impaired diuresis and natriuresis and lowered blood pressure in SHRs, accompanied by reducing ACE and AT1R protein expression and increasing AT2R, ACE2, and MasR expression in the kidneys of SHRs. In addition, these changes were associated with the reduction of ER stress and oxidative stress. Conclusions These results suggest that the imbalance of renal RAS components was associated with increased ER stress in SHRs. Inhibition of ER stress with 4-PBA reversed the imbalance of renal RAS components and restored the impaired diuresis and natriuresis, which, at least in part, explains the blood pressure-lowering effects of 4-PBA in hypertension.

Chronic infusion of ELABELA alleviates vascular remodeling in spontaneously hypertensive rats via anti-inflammatory, anti-oxidative and anti-proliferative effects.

Inflammatory activation and oxidative stress promote the proliferation of vascular smooth muscle cells (VSMCs), which accounts for pathological vascular remodeling in hypertension. ELABELA (ELA) is the second endogenous ligand for angiotensin receptor-like 1 (APJ) receptorthat has been discovered thus far. In this study, we investigated whether ELA regulated VSMC proliferation and vascular remodeling in spontaneously hypertensive rats (SHRs). We showed that compared to that in Wistar-Kyoto rats (WKYs), ELA expression was markedly decreased in the VSMCs of SHRs. Exogenous ELA-21 significantly inhibited inflammatory cytokines and NADPH oxidase 1 expression, reactive oxygen species production and VSMC proliferation and increased the nuclear translocation of nuclear factor erythroid 2-related factor (Nrf2) in VSMCs. Osmotic minipump infusion of exogenous ELA-21 in SHRs for 4 weeks significantly decreased diastolic blood pressure, alleviated vascular remodeling and ameliorated vascular inflammation and oxidative stress in SHRs. In VSMCs of WKY, angiotensin II (Ang II)-induced inflammatory activation, oxidative stress and VSMC proliferation were attenuated by pretreatment with exogenous ELA-21 but were exacerbated by ELA knockdown. Moreover, ELA-21 inhibited the expression of matrix metalloproteinase 2 and 9 in both SHR-VSMCs and Ang II-treated WKY-VSMCs. We further revealed that exogenous ELA-21-induced inhibition of proliferation and PI3K/Akt signaling were amplified by the PI3K/Akt inhibitor LY294002, while the APJ receptor antagonist F13A abolished ELA-21-induced PI3K/Akt inhibition and Nrf2 activation in VSMCs. In conclusion, we demonstrate that ELA-21 alleviates vascular remodeling through anti-inflammatory, anti-oxidative and anti-proliferative effects in SHRs, indicating that ELA-21 may be a therapeutic agent for treating hypertension.

The effects of oat ingredients on blood pressure in spontaneously hypertensive rats.

This study investigated the blood pressure (BP) lowering and cardioprotective effects of oat avenanthramide C and beta-glucan alone or in combination in spontaneously hypertensive rats (SHR). Five-weeks-old male SHR and Wistar-Kyoto rats received vehicle, avenanthramide C and beta-glucan alone or a combination of avenanthramide C and beta-glucan via gavage for 15weeks. BP was measured at 0, 10, and 15weeks of treatment. Echocardiography was performed at 15weeks of treatment. Oxidative stress and inflammation were also measured. Beta-glucan alone prevented the increase in systolic and diastolic BP in SHR, but avenanthramide C alone or the combination did not prevent the increase in systolic and diastolic BP. SHRs treated with beta-glucan and not avenanthramide C or the combination reduced isovolumetric relaxation time when compared to SHR treated with vehicle. Beta-glucan and avenanthramide C decreased the levels of malondialdehyde, a marker of oxidative stress in SHR. In conclusion, beta-glucan is a potential antihypertensive agent that may alleviate cardiovascular abnormalities. PRACTICAL APPLICATIONS: Oats products are believed to contain many bioactives that possess potential beneficial properties against chronic diseases. Specifically, oats beta-glucan has been well-established for its efficacy in positively modulating the risk factor for dyslipidemia. This preliminary in vivo study shows that beta-glucan is a potential antihypertensive agent that may alleviate cardiac dysfunction as well. In light of the current findings, further human studies may establish the efficacy of oats beta-glucan in hypertensive patients alongside the current antihypertensive medications. This novel attribute established via rigorous studies may provide an impetus for oats products and oats industry. Last but not least, it will also help improve the cardiovascular disease burden in a cost-effective way.

Protective effect of antioxidant Tempol on cardiac stem cells in chronic pressure overload hypertrophy

AimsCardiac hypertrophy, an independent risk factor for cardiac failure; is associated with oxidative stress. Decline in the proportion of healthy cardiac stem cells (CSCs), possibly mediated by oxidative stress can lead to cardiac failure. The present study was carried out to examine the hypothesis that reduction of oxidative stress restores CSC efficiency and prevents progressive cardiac remodelling. Materials and methodsSix-month old Spontaneously hypertensive rats (SHR) were supplemented with the antioxidant Tempol (20 mg/kg/day) for 14 days. The effect of Tempol on blood pressure and heart were assessed in SHR. Cardiac stem cells were isolated from atrial explants and expanded in culture for assessment of stem cell characteristics. Intracellular reactive oxygen species (ROS), proliferation, migration and senescence were evaluated in cultured atrial CSCs. Key findingsTempol treatment reduced blood pressure, regressed cardiac hypertrophy and reduced oxidative stress in SHR. Compared to Wistar rat, the efficiency of CSCs was significantly compromised in SHR. Tempol reduced intracellular ROS and restored migration potential and proliferative capacity along with reduction of senescent CSCs and expression of senescence proteins p16ink4a and p21. SignificanceRestoration of functional efficiency of CSCs by antioxidants signifies the role of oxidative stress in deterioration of stem cell attributes in the hypertrophic heart. The observations envisage the use of antioxidants as adjuvant medication for maintaining a healthy stem cell population, which can in-turn prevent progressive cardiac remodelling, a major determinant of cardiac failure.

Hydrogen sulfide improves endothelial dysfunction through inhibition of the vicious circle of NLRP3 and oxidative stress in spontaneously hypertensive rats

AimsThe aim of this study is to elucidate whether the inhibition of inflammasome and oxidative stress mediated the ameliorative effect of hydrogen sulfide (H2S) on endothelial dysfunction in hypertension.MethodsSpontaneously hypertensive rats (SHR) and normotensive Wistar kyoto rats (WKY) were injected with 100 μM NaSH intraperitoneally daily for 16 weeks. Systolic blood pressure (SBP) and plasma MDA and IL‐1β were measured. Renal vascular was used to determine endothelial dependent contraction (EDC) and endothelial dependent relaxation (EDR). Protein levels were detected by Western blot. Human umbilical vein endothelial cells (HUVECs) were used to confirm the protective role of H2S against AngII‐induced cell injury.ResultsExogenous NaSH administration significantly lowered SBP and ameliorated damaged EDR and EDC. H2S reduced the activation of NLRP3 inflammasome and oxidative stress in SHR. The endothelial protective and anti‐oxidant effect of H2S was abolished by LPS, an inducer of NLRP3 inflammasome. In HUVECs, H2S significantly ameliorated AngII‐induced cellular impairment, NLRP3 inflammasome and ROS generation. After knocking down of Nrf2, the above protective effect of H2S was abolished.ConclusionHydrogen sulfide could inhibit vicious circle of oxidative stress and inflammation, resulted in improved endothelial function and ameliorated hypertension. Our results throw light on the crucial role of H2S in regulation of endothelial function, which might provide the new target for treatment of hypertension.Support or Funding InformationThis work was supported by the National Natural Science Foundation of China (31171098, 31671185), the Specialized Research Fund for the Doctoral Program of Higher Education of China (20121323110008), the Hebei Province for Innovation Talents Support Plan (LJRC017) and the Natural Science Foundation of Hebei (C2012206063, H2016206264).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Highly potent antioxidant Olea europaea L. leaf extract affects carotid and renal haemodynamics in experimental hypertension: The role of oleuropein.

Haemodynamic alterations in carotid and renal arteries are associated with the severity of target organ damage in patients with hypertension. Dietary habits, such as the Mediterranean diet, regulate blood pressure and oxidative stress, thus reduce the mortality rate due to cardiovascular diseases. In this study, our aim was to evaluate the reducing activity, antioxidant capacity and metal chelating ability of standardized Olea europaea L. leaf extract (OLE), and to test its (5, 25, 50 mg/kg) acute in vivo effects, as well as oleuropein's (OP, 10 mg/kg) on oxidative stress, carotid, renal and systemic haemodynamic parameters (blood pressure, heart rate, cardiac output, peripheral resistance) in spontaneously hypertensive rats (SHR). OLE has a higher antioxidative capacity than BHT, higher reducing ability than vitamin C, and 23 times lower capacity for metal ion chelation than EDTA. All three doses of OLE, and OP, improved oxidative stress in SHR. OLE5 improved carotid and renal haemodynamics, without significant effects on systemic haemodynamics. Two different mechanisms of antihypertensive responses to OLE were observed, OLE25 was most effective in reducing cardiovascular risks by improving systemic and regional (carotid and renal) haemodynamics, peripheral and regional vascular resistance. OLE50 causes the improvement of blood pressure and cardiac performances, but tends to retain elevated vascular resistance, therefore, reducing the inflow of blood into the brain and kidneys of the SHR. The OP did not alter systemic or regional haemodynamics, suggesting others constituents responsible for changes of cardiac function, as well as carotid and renal haemodynamics in response to OLE50.

Short-Term Treatment with Esmolol Reverses Left Ventricular Hypertrophy in Adult Spontaneously Hypertensive Rats via Inhibition of Akt/NF-κB and NFATc4.

Our group has previously demonstrated that short-term treatment with esmolol reduces left ventricular hypertrophy (LVH) in spontaneously hypertensive rats (SHRs). The present study aimed to assess the molecular mechanisms related to this effect. Fourteen-month-old male SHRs were treated intravenously with saline as vehicle (SHR) or esmolol (SHR-E) (300 μg/kg/min). Age-matched vehicle-treated male Wistar-Kyoto (WKY) rats served as controls. After 48 hours of treatment, the hearts were harvested and left ventricular tissue was separated and processed for Western blot analysis to determine the levels of Akt, NF-κB, NFATc4, Creb1, Serca2a, Erk1/2, and Sapk/Jnk. Biomarkers of oxidative stress, such as catalase, protein carbonyls, total thiols, and total antioxidant capacity were evaluated. Esmolol reversed the levels of p-NFATc4, p-Akt, and p-NF-κB in SHRs to the phospholevels of these proteins in WKY rats without modifying p-Erk1/2, p-Sapk/Jnk, p-Creb1, or Serca2a in SHR. Compared with SHR, esmolol increased catalase activity and reduced protein carbonyls without modifying total thiols or total antioxidant capacity. Short-term treatment with esmolol reverses LVH in aged SHRs by downregulation of Akt/NF-κB and NFATc4 activity. Esmolol treatment also increases catalase activity and reduces oxidative stress in SHRs with LVH.

Effects of Chronic Exposure to Mercury on Angiotensin-Converting Enzyme Activity and Oxidative Stress in Normotensive and Hypertensive Rats.

BackgroundMercury's deleterious effects are associated with increased cardiovascular risk.ObjectiveTo determine whether chronic exposure to inorganic mercury increases the activity of angiotensin-converting enzyme and its relationship with oxidative stress in several organs and tissues.MethodsWe studied male Wistar and spontaneously hypertensive rats (SHR) (3-month-old) exposed or not to HgCl2 for 30 days. At the end of treatment, we investigated the following: changes in body weight, hemodynamic parameters, angiotensin-converting enzyme (ACE) activity and oxidative stress in the heart, aorta, lung, brain and kidney in hypertensive compared to normotensive animals. A value of p < 0.05 was considered significant.ResultsChronic exposure to HgCl2 did not affect weight gain in either group. Systolic blood pressure, measured weekly, did not increase in Wistar rats but showed a small increase in SHR rats. We also observed increases in left ventricular end-diastolic pressure and ACE activity in the plasma and hearts of normotensive rats. In the SHR+Hg group, ACE activity increased in plasma but decreased in kidney, lung, heart, brain and aorta. Oxidative stress was assessed indirectly by malondialdehyde (MDA) production, which increased in Hg-treated rats in both plasma and heart. In the SHR+Hg group, MDA increased in heart and aorta and decreased in lungs and brain.ConclusionThese results suggest that chronic exposure to inorganic mercury aggravates hypertension and produces more expressive changes in ACE activity and oxidative stress in SHRs. Such exposure affects the cardiovascular system, representing a risk factor for the development of cardiovascular disorders in normotensive rats and worsening of pre-existing risks for hypertension.

Gallic Acid Reduces Blood Pressure and Attenuates Oxidative Stress and Cardiac Hypertrophy in Spontaneously Hypertensive Rats

Gallic acid (GA) has been reported to have beneficial effects on cancer, vascular calcification, and diabetes-induced myocardial dysfunction. We hypothesized that GA controls hypertension via oxidative stress response regulation in an animal model for essential hypertension. Spontaneously hypertensive rats (SHRs) were administered GA for 16 weeks. GA treatment lowered elevated systolic blood pressure in SHRs through the inhibition of vascular contractility and components of the renin-angiotensin II system. In addition, GA administration reduced aortic wall thickness and body weight in SHRs. In SHRs, GA attenuated left ventricular hypertrophy and reduced the expression of cardiac-specific transcription factors. NADPH oxidase 2 (Nox2) and GATA4 mRNA expression was induced in SHR hearts and angiotensin II-treated H9c2 cells; this expression was downregulated by GA treatment. Nox2 promoter activity was increased by the synergistic action of GATA4 and Nkx2-5. GA seems to regulate oxidative stress by inhibiting the DNA binding activity of GATA4 in the rat Nox2 promoter. GA reduced the GATA4-induced Nox activity in SHRs and angiotensin II-treated H9c2 cells. GA administration reduced the elevation of malondialdehyde levels in heart tissue obtained from SHRs. These findings suggest that GA is a potential therapeutic agent for treating cardiac hypertrophy and oxidative stress in SHRs.

Novel mechanism within the paraventricular nucleus reduces both blood pressure and hypothalamic pituitary-adrenal axis responses to acute stress.

Macrophage migration inhibitory factor (MIF) counteracts pressor effects of angiotensin II (ANG II) in the paraventricular nucleus of the hypothalamus (PVN) in normotensive rats, but this mechanism is absent in spontaneously hypertensive rats (SHRs) due to a lack of MIF in PVN neurons. Since endogenous ANG II in the PVN modulates stress reactivity, we tested the hypothesis that replacement of MIF in PVN neurons would reduce baseline blood pressure and inhibit stress-induced increases in blood pressure and plasma corticosterone in adult male SHRs. Radiotelemetry transmitters were implanted to measure blood pressure, and then an adeno-associated viral vector expressing either enhanced green fluorescent protein (GFP) or MIF was injected bilaterally into the PVN. Cardiovascular responses to a 15-min water stress (1-cm deep, 25°C) and a 60-min restraint stress were evaluated 3-4 wk later. MIF treatment in the PVN attenuated average restraint-induced increases in blood pressure (37.4 ± 2.0 and 27.6 ± 3.5 mmHg in GFP and MIF groups, respectively, P < 0.05) and corticosterone (42 ± 2 and 36 ± 3 μg/dl in GFP and MIF groups, respectively, P < 0.05). MIF treatment in the PVN also reduced stress-induced elevations in the number of c-Fos-positive cells in the rostral ventrolateral medulla (71 ± 5 in GFP and 47 ± 5 in MIF SHRs, P < 0.01) and corticotropin-releasing factor mRNA expression in the PVN. However, MIF had no significant effects on the cardiovascular responses to water stress in SHRs or to either stress in Sprague-Dawley rats. Therefore, viral vector-mediated restoration of MIF in PVN neurons of SHRs attenuates blood pressure and hypothalamic pituitary adrenal axis responses to stress.

Inhibition of Endoplasmic Reticulum Stress Normalizes Augmented Myogenic Response in Coronary Arteris of the Spontaneously Hypertensive Rats

Endoplasmic reticulum (ER) stress has been shown to play a critical role in the pathogenesis of cardiovascular complications. However, the role and mechanisms of ER stress in hypertension remain unclear. Thus, we hypothesized that enhanced ER stress contributes to the maintenance of hypertension in spontaneously hypertensive rats (SHRs). We investigated this hypothesis by suppressing ER stress in SHRs and their normotensive controls, Wistar‐Kyoto rats (WKYs) with ER stress inhibitor (taurine‐conjugated ursodeoxycholic acid, TUDCA). Sixteen‐week old male SHRs and WKYs were treated with TUDCA (100 mg/kg/day, IP injection) or PBS (control, 300 μl/day, IP injection) for two weeks. There was a decrease in systolic blood pressure in SHR treated with TUDCA but not in SHR treated with PBS. The myogenic response is potentiated in coronary arteries of SHRs compared to WKYs. Interestingly, treatment of ER stress inhibitor normalized myogenic responses in SHRs. These data were associated with an increase in ER stress marker expression (Bip, CHOP, ATF6, XBP‐1, and phosphorylated‐eIF2α) in coronary arteries of SHRs, which were reduced by TUDCA treatment. In conclusion, ER stress inhibition decreases systolic blood pressure and normalizes myogenic response in SHRs. Moreover, ER stress inhibition reduces expression of ER stress markers. Therefore, we suggest that ER stress could be a potential target for hypertension.

Coconut oil supplementation and physical exercise improves baroreflex sensitivity and oxidative stress in hypertensive rats.

The hypothesis that oral supplementation with virgin coconut oil (Cocos nucifera L.) and exercise training would improve impaired baroreflex sensitivity (BRS) and reduce oxidative stress in spontaneously hypertensive rats (SHR) was tested. Adult male SHR and Wistar Kyoto rats (WKY) were divided into 5 groups: WKY + saline (n = 8); SHR + saline (n = 8); SHR + coconut oil (2 mL·day(-1), n = 8); SHR + trained (n = 8); and SHR + trained + coconut oil (n = 8). Mean arterial pressure (MAP) was recorded and BRS was tested using phenylephrine (8 μg/kg, intravenous) and sodium nitroprusside (25 μg·kg(-1), intravenous). Oxidative stress was measured using dihydroethidium in heart and aorta. SHR + saline, SHR + coconut oil, and SHR + trained group showed higher MAP compared with WKY + saline (175 ± 6, 148 ± 6, 147 ± 7 vs. 113 ± 2 mm Hg; p < 0.05). SHR + coconut oil, SHR + trained group, and SHR + trained + coconut oil groups presented lower MAP compared with SHR + saline group (148 ± 6, 147 ± 7, 134 ± 8 vs. 175 ± 6 mm Hg; p < 0.05). Coconut oil combined with exercise training improved BRS in SHR compared with SHR + saline group (-2.47 ± 0.3 vs. -1.39 ± 0.09 beats·min(-1)·mm Hg(-1); p < 0.05). SHR + saline group showed higher superoxide levels when compared with WKY + saline (774 ± 31 vs. 634 ± 19 arbitrary units (AU), respectively; p < 0.05). SHR + trained + coconut oil group presented reduced oxidative stress compared with SHR + saline in heart (622 ± 16 vs. 774 ± 31 AU, p < 0.05). In aorta, coconut oil reduced oxidative stress in SHR compared with SHR + saline group (454 ± 33 vs. 689 ± 29 AU, p < 0.05). Oral supplementation with coconut oil combined with exercise training improved impaired BRS and reduced oxidative stress in SHR.

Amlodipine Increased Endothelial Nitric Oxide and Decreased Nitroxidative Stress Disproportionately to Blood Pressure Changes

Clinical trials have shown that amlodipine reduces cardiovascular events at a rate that is not predicted by changes in brachial arterial pressure alone. These findings may be explained, in part, by the pleiotropic effects of amlodipine on endothelial cell (EC) function. In this study, we elucidated the effect of amlodipine on nitric oxide (NO) bioavailability and cytotoxic peroxynitrite (ONOO(-)) and blood pressure (BP). Spontaneously hypertensive rats (SHRs) were treated with vehicle or amlodipine (5 mg/kg/day) for 8 weeks and compared with untreated, baseline rats. NO and ONOO(-) release from aortic and glomerular ECs were measured ex vivo using amperometric nanosensors following maximal stimulation with calcium ionophore. BP was measured using the tail-cuff method. As compared with baseline, vehicle treatment had reduced aortic endothelial NO release from 157 ± 11 nM to 55 ± 6 nM and increased ONOO(-) from 69 ± 7 nM to 156 ± 19 nM. The NO/ONOO(-) ratio, a comprehensive measurement of eNOS function, decreased from 2.3 ± 0.3 to 0.3 ± 0.1. Compared with vehicle, amlodipine treatment restored NO to 101 ± 3 nM, decreased ONOO(-) to 50 ± 4 nM, and increased the NO/ONOO(-) ratio to 2.0 ± 0.2, a level similar to baseline. Similar changes were observed for glomerular ECs. Mean arterial blood pressure increased from 149 ± 3 mm Hg (baseline) to 174 ± 1 mm Hg (vehicle). Amlodipine slightly, but significantly, decreased mean arterial blood pressure to 167 ± 3 mm Hg vs. vehicle treatment. Amlodipine increased NO bioavailability and decreased nitroxidative stress in SHRs with EC dysfunction disproportionately to BP changes. These direct, vascular effects of amlodipine on EC function may contribute to reduced risk for atherothrombotic events as observed in clinical trials.

Administration of Telmisartan Reduced Systolic Blood Pressure and Oxidative Stress Probably Through the Activation of PI3K/Akt/eNOS Pathway and NO Release in Spontaneously Hypertensive Rats

We investigated the effects of telmisartan, the blocker of angiotensin II receptor 1, on the regulation of systolic blood pressure (SBP) and oxidative stress through endothelial nitric oxide (NO) release in spontaneously hypertensive rats (SHRs). SHRs randomly received placebo, oral feeding of telmisartan (5 mg/kg or 10 mg/kg) every day and Wistar-Kyoto rats (WKYs) served as normotensive control. The SBP of rat was measured before and weekly thereafter. After a total of 8-week treatment, rats were killed for experimental measurements. Parameters that subject to measurements in isolated aorta endothelial cells include: NO concentration, protein expression levels of angiotensin II receptor 1, nitrotyrosine, 8-isoprostane, SOD, PI3K, Akt, AMPK and eNOS. In addition, L-NMMA, a general inhibitor of nitric oxide synthase, was also applied to test the inhibition of NO concentration. We found that SBPs were significantly lower in telmisartan therapy group than in placebo treated hypertensive rats and WKYs (p<0.05). The NO concentration was significantly higher in telmisartan-treated group with increased activity of the PI3K/Akt pathway and activated eNOS signaling. Blockade of Akt activity reversed such effects. Activation of AMPK also contributed to the phosphorylation of eNOS. L-NMMA treatment reduced less NO concentration in SHR rats than the telmisartan co-treated groups. Oxidative stress in SHRs was also attenuated by telmisartan administration, shown by reduced formation of nitrotyrosine, 8-isoprostane, and recovered SOD protein level. Telmisartan enhanced NO release by activating the PI3K/Akt system, AMPK phosphorylation and eNOS expression, which attenuated the blood pressure and oxidative stress in SHRs.

2,4,3′,5′‐Tetramethoxystilbene Reduces Blood Pressure and Associated Cardiac Fibrosis Via Inhibition of Cytochrome P450 1B1 and Decreased Oxidative Stress in SHR

Previously, we reported that inhibition of cytochrome P450 (CYP) 1B1 activity with 2,4,3′,5′‐tetramethoxystilbene (TMS) reversed angiotensin II‐ and deoxycoticosterone acetate salt‐induced hypertension and associated pathophysiology in rats. This study was conducted to investigate the effect of TMS on blood pressure (BP), and associated cardiac fibrosis and oxidative stress in male, normotensive Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR). TMS (600 μg/kg, i.p. daily for 6 weeks) beginning at 8 weeks of age, did not alter systolic BP in WKY, but decreased it in SHR (207 ± 7 mmHg vehicle, vs. 129 ± 2 mmHg TMS, P &lt; 0.05). Cardiac CYP1B1 activity that was increased in SHR was reduced by TMS treatment. Cardiac fibrosis, as indicated by increased collagen deposition and myofibroblast accumulation, was observed in SHR, which was prevented by TMS. SHR had increased cardiac levels of reactive oxygen species, increased NADPH oxidase activity, and increased plasma levels of H2O2, NOx, and TBARS that were prevented by TMS, suggesting that in addition to inhibiting CYP1B1 activity and limiting cardiac damage, TMS regulates cardiac oxidative stress in SHR. These data suggest that CYP1B1 contributes to increased BP, cardiac damage, and associated oxidative stress in SHR. Moreover, CYP1B1 could serve as a novel target for the development of drugs such as TMS to treat cardiac disease associated with hypertension.

Quercetin Improves Baroreflex Sensitivity in Spontaneously Hypertensive Rats

Quercetin is a well-known antioxidant. Here, we investigated the effects of treatment with quercetin on mean arterial pressure (MAP), heart rate (HR) and baroreflex sensitivity (BRS) in spontaneously hypertensive rats (SHR). SHR and their controls (WKY) were orally treated with quercetin (2, 10 or 25 mg/kg/day) or saline for seven days. On the 8th day, MAP and HR were recorded. BRS was tested using phenylephrine (8 mg/kg, i.v.) and sodium nitroprusside (25 mg/kg, i.v.). Oxidative stress was measured by tiobarbituric acid reactive species assay. The doses of 10 (n = 8) and 25 mg/kg (n = 8) were able to decrease the MAP in SHR (n = 9) (163 ± 4 and 156 ± 5 vs. 173 ± 6, respectively, p < 0.05) but not in WKY (117 ± 1 and 118 ± 2 vs. 113 ± 1, respectively, p < 0.05). The dose of 25 mg/kg/day increased the sensitivity of parasympathetic component of the baroreflex (−2.47 ± 0.31 vs. −1.25 ± 0.8 bpm/mmHg) and decreased serum oxidative stress in SHR (2.04 ± 0.17 vs. 3.22 ± 0.37 nmol/mL, n = 6). Our data suggest that treatment with quercetin reduces hypertension and improves BRS in SHR via reduction in oxidative stress.

Liver growth factor treatment reverses vascular and plasmatic oxidative stress in spontaneously hypertensive rats

Liver growth factor (LGF) is an albumin-bilirubin complex with antioxidant actions in vitro. In spontaneously hypertensive rats (SHRs), short LGF treatment exerts antihypertensive and antifibrotic effects. We aimed to determine if LGF treatment (4 i.p. injections, 4.5 μg/rat over 12 days) reduces oxidative stress in SHRs using Wistar-Kyoto (WKY) as control strain. We assessed the following: plasma oxidative stress biomarkers [protein-bound malondialdehyde (MDA); protein carbonyls and advanced glycation end products (AGEs)]; superoxide anion basal production in carotid artery-derived vascular smooth muscle cells (VSMCs) detected by dihydroethidium and confocal microscopy; and expression (western blot) and activities (spectroscopic methods) of NADPH and xanthine oxidases, CuZn, Mn and extracellular superoxide dismutases (SODs) and catalase in carotid arteries. LGF treatment had the following effects: reversed the increase in plasma MDA and protein carbonyls and VSMC superoxide anion levels observed in SHRs, without any effect on WKY strain; reversed the alterations in SHR vascular p22phox expression as well as NADPH oxidase, xanthine oxidase and catalase activities; had no effect on vascular CuZn-SOD and Mn-SOD expression or total SOD activity; and reversed the elevation in SHR vascular glycated/free extracellular-SOD expression ratio and plasma glucose without changes in plasma AGEs. LGF treatment of SHRs normalizes the level of plasma oxidative stress biomarkers through a reduction of vascular superoxide anion produced by NADPH and xanthine oxidases. These effects might be linked to the cardiovascular regenerative actions of LGF.

Scavenging of NADPH oxidase‐derived superoxide anions improves depressed baroreflex sensitivity in spontaneously hypertensive rats

In pathological conditions, such as hypertension, there is impairment in the autonomic control of blood pressure resulting in changes in baroreflex sensitivity. In the present study we tested the hypothesis that acute superoxide scavenging would restore the depressed baroreflex sensitivity (BRS) in spontaneously hypertensive rats (SHR). Male 10-week-old SHR (n=14) and their controls (Wistar-Kyoto (WKY) rats; n=14) underwent femoral artery and vein catheterization for conscious blood pressure recording and drug administration. The BRS was obtained by the drug-induced method using phenylephrine (8μg/kg, i.v.) and sodium nitroprusside (25μg/kg, i.v.) before and after the administration of tiron (30mg/kg, i.v.), a superoxide dismutase mimetic, or apocynin (30μg/kg), an NADPH oxidase inhibitor. Spontaneously hypertensive rats was significantly hypertensive compared with WKY rats (160±7 vs 105±2mmHg, respectively). However, there was no significant difference in heart rate between the two groups (388±10 vs 370±20b.p.m.). In addition, SHR exhibited a diminished BRS compared with WKY rats (-1.34±0.11 vs -2.91±0.20b.p.m./mmHg, respectively). Administration of tiron improved BRS in SHR (from -1.34±0.11 to 2.26±0.21b.p.m./mmHg), as did apocynin (to -2.14±0.23b.p.m./mmHg). Serum samples from SHR (n=20) and WKY rats (n=20) were collected for thiobarbituric acid-reactive substances assays before and after tiron or apocynin to confirm the reduction in oxidative stress. There was considerably greater oxidative stress in SHR compared with WKY rats (36.2±3.0 vs 13.3±2.6nmol/L, respectively). Both apocynin and tiron treatment reduced the oxidative stress in SHR (from 36.2±3.0 to 21.5±3.0nmol/L and from 37.2±3.9 to 21.9±1.6nmol/L, respectively). The data suggest that acute scavenging of NADPH oxidase-derived superoxide improves baroreflex sensitivity in SHR.

Influence of acute hypoxia combined with nitrous oxide on cardiovascular variability in conscious hypertensive rats

Anesthetics have been reported to depress autonomic nervous system (ANS) responses to hypoxia. The mechanisms by which cardiovascular variability responds to acute progressive hypoxia (APH) under nitrous oxide (N 2O) inhalation, however, remain unclear. Additionally, the effect of hypertension on ANS responses in such cases has not been fully clarified. The present study examined the influence of APH (10% O 2) under 60% N 2O inhalation on cardiovascular variability in conscious, spontaneously hypertensive rats (SHR). Twenty-seven male SHR were randomly assigned to 3 treatment groups receiving N 2O inhalation alone, APH stress alone or APH stress under N 2O inhalation, using Wistar Kyoto rats (WKY) or non-N 2O inhalation rats as controls. Systolic blood pressure (SBP) and heart rate (HR) variability were evaluated time-dependently using the wavelet method. While inhalation of N 2O alone induced more powerful sympathomimetic actions in SHR than in WKY, circulatory and parasympathetic reactions were weaker. APH stress alone evoked significant inhibition of cardiac parasympathetic activity from immediately after exposure to hypoxic stress in SHR in contrast to WKY, facilitating tachycardia. This inhibition of parasympathetic activity in SHR continued without coupled changes in sympathetic activity. In SHR, APH under N 2O inhalation decreased SBP and sympathetic activity more prominently and earlier than APH alone, and earlier than APH under N 2O inhalation in WKY. Additionally, APH under N 2O inhalation inhibited cardiac parasympathetic activity in SHR as compared to APH stress alone. In conclusion, APH under N 2O inhalation in SHR potentially results in exacerbation of circulatory suppression from the earlier hypoxic phase, compared with non-N 2O inhalation.