HPA041603 consistently detected SGLT2 in rodent and human kidney tissues, whereas other antibodies showed limited specificity. Knockout-based antibody validation and optimized experimental conditions are essential for accurate interpretation of SGLT2 protein expression in experimental and translational studies.

Salt-Sensitive hypertension (SS-HTN) is associated with an increase in immune cells and pro-inflammatory cytokines, such as IL-6. Additionally, consuming a high salt (HS) diet contributes to the development of SS-HTN, as well as induces renal immune cell activation. We previously showed that intra-renal IL-6 is increased in SS-HTN and can increase blood pressure via reduced sodium excretion. However, no data show a mechanistic link between dietary HS in SS-HTN and inflammation. We hypothesize that HS increases renal osmolarity and IL-6 levels, which precedes the development of SS-HTN. Wt (male) mice were given L-NAME (0.5mg/mL, 2 wks) and then washed-out (1-2wks, Fig 1A ). Mice in early phase (early salt-sensitive, eSS) were then given 1% saline + normal chow (NC) or HS chow (4%) + tap water for 2 days. To develop SS-HTN, mice were given HS for the remainder of the study (SS-HTN, Fig 1A ). Control mice were given NC or HS only. Kidneys were isolated and dissected (cortex + outer medulla) for osmolarity or used for RT-PCR. Serum was used for cytokine analysis (Meso-Scale Discovery). To determine how and where immune cells may be exposed to HS, we assessed the interstitial osmolarity of the renal cortex + outer medulla in both models of eSS. We observed a significant increase in interstitial osmolarity at the eSS stage following 2 days of 1% saline or 4% HS chow, as compared to NC ( Fig 1B ). This increase was not present during SS-HTN. This corresponds with early increases in serum IL-6 levels (1.82±0.3 pg/mL eSS vs. 0.81±0.14pg/mL HS vs . 1.1±0.27pg/mL NC; p<0.05) and renal CD8 + T-cells. We then assessed kidney injury molecule 1 (KIM-1) mRNA levels and saw the greatest increase was also at eSS (4%) (DCt 9.58±0.17 eSS vs. DCt 9.58±0.17, p<0.01). Here, we are the first to show that salt-sensitivity (eSS) promotes increased osmolarity accumulation in the renal interstitium. This also corresponds with the greatest increase in IL-6 and KIM-1, all preceding the development of SS-HTN. These data suggest that HS-induced increases in IL-6 and KIM-1, via increased renal osmolarity, may contribute to the early development of SS-HTN.

Background: Glucagon-like peptide-1 receptor agonists (GLP-1RAs) are widely used for metabolic disease, but their effects on hypertension remain incompletely understood. This study tested the hypothesis that semaglutide, a long-acting GLP-1RA, attenuates blood pressure elevation and cardiometabolic abnormalities in a preclinical model of diet-induced hypertension. Methods: Male Dahl Salt-Sensitive rats (n=17) were fed a control diet (CD; 10% kcal from fat) or a high-fat diet (HFD; 60% kcal from fat) beginning at 4 weeks of age. After 21 weeks, rats received daily subcutaneous semaglutide (0.1 mg/kg) or vehicle for 28 days. Radiotelemetry enabled continuous monitoring of mean arterial pressure (MAP), heart rate, activity, and core temperature. Metabolic endpoints included body weight, food intake, fasting glucose, blood chemistry (renal, hepatic, lipid panels), and micro-CT imaging of adiposity. Results: HFD-fed rats exhibited significantly higher body weight (CD: 446.0 ± 4.7 g vs. HFD: 466 ± 3.2 g; p<0.05) and MAP (CD: 131.4 ± 0.7 mmHg vs. HFD: 142.5 ± 2.3 mmHg; p<0.05) than CD-fed controls. Semaglutide significantly reduced body weight in HFD rats (-11.2 ± 5.2 g from baseline; p<0.05) and decreased MAP in both CD and HFD groups (-8.4 ± 1.1 mmHg and -8.4 ± 0.9 mmHg, respectively; p<0.05). Treatment elicited a sustained heart rate increase (peak: +97.4 BPM in CD; +87.8 BPM in HFD; p<0.05), along with transient reductions in food/fluid intake and core body temperature. Locomotor activity declined temporarily, consistent with treatment-related malaise. Micro-CT imaging revealed substantial reductions in both visceral and subcutaneous fat depots with semaglutide. Markers of renal and hepatic function remained within normal ranges, though changes in biliary markers warrant further study. Correlation analysis linked reductions in adiposity and body weight to the antihypertensive effect of semaglutide. Conclusions: Semaglutide reduces blood pressure and body weight in a model of diet-induced hypertension. These findings support a role for GLP-1RAs in mitigating obesity-related hypertension via mechanisms beyond glucose lowering. Grant Support: NHLBI P01HL152951.

Hypertension and metabolic conditions like dyslipidemia and diabetes damage multiple organ systems, including the heart, brain and kidneys. Recent studies suggest the potential interplay between metabolic disorders and hypertension which increases morbidity and mortality. In 2025, it's estimated that nearly 47% of US adults have high blood pressure, while over 72% have an unhealthy weight, 42% being obese. Importantly, 30% of hypertensive patients remain hypertensive despite multiple treatments. Thus, there is an urgent need for new therapies. We discovered a novel role of acetylation of mitochondrial proteins in these conditions. Essential hypertension and metabolic disorders are linked to hyperacetylation and inactivation of critical mitochondrial metabolic and antioxidant enzymes, long-chain acyl coenzyme A dehydrogenase (LCAD) and superoxide dismutase 2 (SOD2). The specific role of these pathways is unclear, and understanding these mechanisms is important to develop new therapies. High fat diet and hypertension are linked to mitochondrial lipid peroxidation and formation of highly toxic oxidized lipid products, isolevuglandins (isoLGs). IsoLGs can promote acetylation of LCAD and SOD2 driving metabolic dysfunction and multi-organ damage, however, their roles in organ damage and hypertension are still elusive. We suggest that LCAD and SOD2 acetylation induces metabolic-oxidative stress crosstalk which promotes mitochondrial dysfunction and multi-organ damage. To test this hypothesis, we have developed new mouse models using LCAD K318/322Q acetylation mimetic mice (LCADQ) and SOD2-K68Q acetylation mimetic mice (SOD2Q). Our data show increased oxidative stress as well as impaired fatty acid and glucose metabolism in both LCADQ and SOD2Q mice. Histological analysis showed significant increase in muscularized lung blood vessels, arterial wall thickness, kidney cell injury, loss of brush border and tubule dilatation in the LCADQ mice compared with wildtype. SOD2Q studies showed increased vascular oxidative stress promoting hypertension and vascular deficiency of medium-chain fatty acids supporting impaired fatty acid oxidation in SOD2Q mice. Treatment with mitochondria-targeted isoLGs scavenger mito2HOBA improved vasorelaxation and reduced hypertension in LCADQ mice. Our data strongly support the novel role of mitochondrial acetylation in metabolic-oxidative stress interplay and provides novel therapeutic targets for treatment of these pathological conditions.

Background: Accurate and autonomous blood pressure (BP) monitoring remains a significant challenge in hypertension management. We evaluated a novel subcutaneous pulse-echo ultrasound (US) system designed to calculate arterial pressure waveforms, comparing its performance to intra-arterial (IA) catheter-based reference measurements in an acute preclinical study. Methods: In anesthetized sheep, continuous BP measurements were obtained via an IA catheter in the femoral artery. Simultaneously, 10-second US recordings (n = 38) were collected using a transducer placed near target arteries (carotid and femoral). These signals were processed through a proprietary algorithm to extract pressure waveforms based on arterial diameter changes. Calibration was performed by correlating time-varying diameters with reference pressure values over multiple heartbeats. The resulting calibration parameters were then used to estimate pressure values in subsequent beats. The accuracy of US-derived pressures was assessed by comparing systolic and diastolic values to IA readings using Bland-Altman analysis. Results: System accuracy was quantified by comparing systolic and diastolic pressures derived from US waveforms to IA references across multiple cardiac cycles and repeated measurements. As illustrated in Figure 1, US-derived waveforms (blue) closely resemble IA reference waveforms (red) and are capable of capturing key features such as the systolic upstroke, dicrotic notch, and diastolic decay. The Bland-Altman analysis (Figure 2) demonstrated strong agreement across a range of pressures, with mean absolute differences of 2.0 mmHg (systolic) and 2.2 mmHg (diastolic). Conclusions: The subcutaneous ultrasound BP system provides accurate and repeatable systolic and diastolic pressure measurements compared to the gold-standard IA catheter. These findings support the feasibility of using subcutaneously implanted ultrasound for BP monitoring and suggest potential for future diagnostic capabilities. Ongoing work will address chronic implantation and real-time algorithm optimization for use in freely moving subjects.

In the brain, astrocytes play a crucial role in maintaining synaptic plasticity. Recent studies have shown that perineuronal nets (PNNs) are essential for astrocyte function by supporting the integrity of the tripartite synapse in the mediobasal hypothalamus (MBH), a key brain region regulating metabolic physiology. However, it remains unclear whether MBH PNNs are dysregulated in the context of long-term high-fat diet (HFD) exposure and whether astrogliosis contributes to altered PNN formation, particularly in obesity-related hypertension. We hypothesized that long-term HFD induces PNN remodeling in association with astrogliosis. To test this, C57BL/6J mice were fed a high-fat (60% kcal from fat, n=3) or control diet (n=4) for 40 weeks. Continuous blood pressure was monitored in week 40 using telemetry probes. Immunofluorescent labeling of glial fibrillary acidic protein (GFAP), a marker of astrogliosis, and Wisteria floribunda agglutinin conjugated to fluorescein isothiocyanate (WFA-FITC), a marker of PNNs, was performed in the arcuate nucleus (ARC) and the ventromedial hypothalamus (VMH), two key regions in the MBH. In HFD mice, mean arterial pressure (115.09 ± 1.95 mmHg vs. 102.06 ± 1.84 mmHg; p <0.05) and heart rate (633.00 ± 4.84 BPM vs. 583.78 ± 9.44 BPM; p <0.05) were significantly elevated. Notably, HFD mice exhibited lower heart rate variability (8.84 ± 0.9 ms vs. 12.9 ± 0.51 ms; p <0.05), suggesting heightened sympathetic and/or reduced parasympathetic activity. Immunofluorescence analyses revealed a significant reduction in the mean fluorescence intensity (MFI) of PNNs (ARC; 5.53 ± 1.46 vs. 24.64 ± 2.83; p <0.0001, VMH; 13.76 ± 3.12 vs. 39.36 ± 5.81; p =0.0008) in HFD mice. In contrast, GFAP intensity was significantly increased (ARC; 43.06 ± 4.34 vs. 19.75 ± 3.59; p =0.0007, VMH; 10.64 ± 2.02 vs 2.86± 1.36; p =0.02) in HFD mice. Astrogliosis upregulates proteolytic enzymes responsible for PNN degradation. Further correlation analyses demonstrated a significant inverse relationship in GFAP and PNN intensity, ARC ( p =0.003) and VMH ( p =0.05), highlighting a potential role of astrogliosis in PNN remodeling within the MBH. In summary, Obesity-related hypertension is associated with reduced PNN integrity and increased astrogliosis in the MBH, and both phenomena are inversely correlated. These results suggest that the astrocyte-PNN interactions may represent a novel pathway for neurogenic hypertension.

Hypertension affects over 50% of adults and is associated with the overactivation of the paraventricular nucleus of the hypothalamus (PVH), which regulates neurohumoral and sympathetic output. We investigate perineuronal nets (PNNs) as a potential mechanism underlying this overactivation. PNNs are lattice-like extracellular structures surrounding the soma and proximal dendrites of neurons, influencing their firing and circuit plasticity. In addition to PVH, we examine PNN changes in the perifornical area of the anterior hypothalamus (PeFAH), an inhibitory, PNN-rich region adjacent to and potentially projecting to the PVH. We hypothesize that hypertension increases PNNs in the PVH, enhancing excitability, while decreasing PNNs in the PeFAH, reducing its inhibitory output – together contributing to PVH overactivation. To test this, 3–5-month-old male and female C57BL6 mice were implanted with s.c. deoxycorticosterone acetate (DOCA) pellets and given 0.9% NaCl in drinking water. After 21 days of DOCA-salt treatment, perineuronal nets (PNNs) were quantified using the marker Wisteria Floribunda Agglutinin (WFA). In the PVH, DOCA-salt males showed a significant increase in WFA+ area (%) (mean ± SD) (3.636 ± 1.636, n = 9) compared to sham controls (2.497 ± 0.9924, n = 10; p = 0.0255, two-tailed unpaired t-test). This corresponded to a higher PNN number (n/mm 2 ) in treated mice (DOCA: 44.77 ± 11.64; Sham: 35.63 ± 6.198; p = 0.0447). In contrast, females showed no significant change in WFA+ area (DOCA: 4.226 ± 1.386; Sham: 3.966 ± 1.747; p = 0.5532) or PNN number (DOCA: 64.87 ± 16.88; Sham: 71.48 ± 11.70; p = 0.4011), despite elevated blood pressure. In the PeFAH, DOCA-salt males exhibited a reduction in normalized WFA+ area (DOCA: 0.9200 ± 0.0656; Sham: 1.001 ± 0.0927; p = 0.0440), which was not observed in females (DOCA: 0.9518 ± 0.0858; Sham: 1.000 ± 0.0851; p = 0.2953). These results suggest that increased PVH PNNs may activate previously ‘silent’ neurons, while reduced PNNs in the PeFAH may weaken inhibitory output – together contributing to PVH overactivation in male mice during hypertension.

Introduction: Orthostatic hypotension (OH) is a debilitating condition commonly observed in patients with impaired vascular tone upon positional changes, often due to neurogenic disorders, liver disease, hypovolemia, or advanced HF. While fludrocortisone has been associated with adverse outcomes—particularly HF-related complications—compared to midodrine in HF populations, direct comparative data between these two therapies in the general population with OH is seldom. Hypothesis: This study aims to compare cardiovascular and renal outcomes in patients with OH treated with fludrocortisone (without midodrine) versus those treated with midodrine (without fludrocortisone). Methods: A retrospective cohort study was conducted using data from the TriNetX collaborative research network from January 2007 to May 2025. Patients aged 18 or older diagnosed with OH were included. After propensity score matching for demographics, comorbidities, and medications, two cohorts were analyzed: cohort 1 included 33,101 patients treated with fludrocortisone (excluding midodrine), and cohort 2 included 33,101 treated with midodrine (excluding fludrocortisone). Mean age was 62 years; 51% were female and 47% male. Risk ratios (RR) and hazard ratios (HR) with 95% confidence intervals and p-values were calculated using TriNetX. The outcomes assessed included all cause mortality, syncope/collapse, stroke, heart failure (HF), acute myocardial infarction (AMI), atrial fibrillation (AF), and ventricualr fibrillation (VF). Results: Though mortality risk was higher in cohort 1 (26.85% vs. 22.12%, RR 1.214, 95% CI 1.181–1.247; p<0.0001), HR was 0.938 (95% CI 0.91–0.968; p<0.0001). Syncope was more frequent in cohort 1 (25.84% vs. 21.77%, RR 1.187; p<0.0001; HR 1.034; p=0.025), as was stroke (8.09% vs. 7.09%, RR 1.142; HR 0.939; p=0.0375). AMI (6.05% vs. 6.92%, RR 0.873; HR 0.694; p<0.0001), HF (17.81% vs. 22.99%, RR 0.775; HR 0.636; p<0.0001), AF (18.50% vs. 22.54%, RR 0.821; HR 0.695; p<0.0001), and VF (0.34% vs. 0.62%, RR 0.544; HR 0.435) were all lower in cohort 1. AKI/CKD risk was similar (30.43% vs. 30.66%, RR 0.993; p=0.5213), though HR favored fludrocortisone (0.846; p<0.0001). Conclusion: In OH patients, midodrine was linked to lower all-cause mortality risk, but fludrocortisone showed better survival and lower risks of HF, AMI, AF, VF, and cardiovascular mortality. Syncope and cerebrovascular events were more common with fludrocortisone.

Combination antiretroviral therapy has markedly increased life expectancy in people living with HIV (PLWH) and shifted their primary cause of death from opportunistic to cardiovascular disease (CVD). Africans and individual of African Descent (IAD) are disproportionately affected by HIV, and exhibit accelerated development of hypertension and CVD, independent of their HIV status. Polymorphism in the Duffy antigen receptor for chemokines (DARC), which is associated with heightened HIV infectivity and systemic inflammation due to impaired chemokine clearance, is predominantly prevalent in IAD. To investigate how DARC deficiency interacts with HIV to promote CVD, we used mouse models, either deficient in DARC (DARC KO), constitutively expressing HIV-encoded proteins (Tg26 mouse), or expressing the 2 genetic mutations (Tg26-DARC). We hypothesized that HIV-derived proteins and DARC deficiency independently and synergistically contribute to endothelial dysfunction and hypertension. BP measurements and vascular reactivity studies revealed elevated systolic BP and impaired endothelial-dependent relaxation in Tg26 and DARC KO mice. Tg26 mice exhibited high circulating IL-1α and vascular NOX1 expression, and experiments with neutralizing antibody revealed that vascular dysfunction and hypertension were mediated via an IL-1α/NOX1 mechanism. In contrast, DARC KO mice showed reduced circulating IL-1α and vascular NOX1 levels, but displayed elevated plasma TNFα, IL-1β, and IL-6. Furthermore, DARC-regulated chemokines (CCL2, CCL5, CCR5, CCL7, CXCL2, and CXCL3) were significantly upregulated in CD4+ T cells of DARC KO mice. Interestingly, DARC deficiency in Tg26 mice also led to reduced IL-1α and NOX1, with no additive effect on endothelial function and BP. To examine the role of hematopoietic-expressed DARC and immune cell-derived viral proteins in endothelial dysfunction, we performed bone marrow transplants (BMT). Endothelial-dependent relaxation was impaired in resistance arteries of WT mice receiving DARC KO, Tg26, or Tg26 DARC KO bone marrow. Similarly, systolic BP was significantly elevated in these groups compared to WT to WT transplants. IL-1α was selectively increased in Tg26-WT mice, similar to the donor mice. In conclusion, HIV-derived proteins promote vascular dysfunction through IL-1α/NOX1 signaling, while DARC deficiency contributes to vascular dysfunction through chemokine dysregulation without synergistic effects between DARC and viral proteins.

Background: Diuretics are widely used for hypertension but may raise hypokalemia risk. Dietary sodium-to-potassium (Na/K) ratio is key for potassium homeostasis. The DASH (Dietary Approaches to Stop Hypertension) diet, promoting low sodium and high potassium intake, is recommended for blood pressure control, yet its role in modifying diuretic-related electrolyte effects remains underexplored. This study examined (1) the association between diuretic use and hypokalemia and (2) whether dietary Na/K ratio or serum sodium modifies this association in treated hypertensive adults. Methods: We analyzed cross-sectional data from the 2017- 2020 National Health and Nutrition Examination Survey. Hypokalemia was defined as serum potassium <3.5 mEq/L. First-day dietary Na/K ratio was calculated. Logistic regression models estimated hypokalemia odds associated with diuretic use, adjusting sequentially for demographic, clinical, and biochemical factors (serum sodium, dietary Na/K ratio). Effect modification by dietary Na/K ratio and serum sodium was assessed using interaction terms, with marginal effects estimated at 25th, 50th, and 75th percentiles. Results: Among 3,426 U.S. adults treated for hypertension, 16.2% (n = 554) reported diuretic use. Hypokalemia was more prevalent in diuretic users than non-users (10% vs. 4.7%, p < 0.001). In unadjusted analyses ( Table 1) , diuretic use was linked to higher hypokalemia odds (Model 0: OR = 2.28, 95% CI: 1.65–3.16), remaining consistent after adjusting for demographics (Model 1: OR = 2.46, 95% CI: 1.72–3.53), clinical factors (Model 2: OR = 2.64, 95% CI: 1.82–3.83), and serum sodium and dietary Na/K ratio (Model 3: OR = 2.66, 95% CI: 1.80–3.91). In stratified analyses ( Table 2 ), the association strengthened across increasing dietary Na/K ratio percentiles: OR = 2.27 at 25% (0.99), 2.56 at 50% (1.34), and 2.99 at 75% (1.78). In contrast, the association remained stable across serum sodium percentiles: OR = 2.71 at 25% (139 mmol/L), 2.66 at 50% (141mmol/L), and 2.62 at 75% (143mmol/L). Conclusion: Among treated hypertensive adults, diuretics were associated with significantly higher odds of hypokalemia. This risk was more pronounced in individuals with higher dietary Na/K ratios, underscoring the potential protective role of DASH-aligned diets in mitigating electrolyte-related side effects of antihypertensive therapy.