The U.S. national average for the prevalence of hypertension in women aged 35-44 is 10.8%. In southeastern states including Georgia, Mississippi, and South Carolina, the prevalence of hypertension in women of this same age group is 13.5%, 17.8%, and 15.5%, respectively. The increase is mostly observed in Black women from non-metro areas. Thus, a need exists to improve awareness of cardiovascular risk in these communities and to support scientific research to improve the understanding of how sex contributes to differences in cardiovascular risk factors like hypertension. Rural Obese At Risk (ROAR) establishes a Specialized Center of Research Excellence on Sex Differences (SCORE) homed at Augusta University, spanning three Southeast universities. The overall goal is to establish ROAR as a national resource and recognized leader in cardiovascular disease in females, and to change perceptions and awareness surrounding cardiovascular risk in women across the Southeast, particularly in high-risk groups. One objective of ROAR Augusta is to establish a community outreach pipeline of trained scientists to increase awareness of sex as a biological variable (SABV), promote optimal cardiovascular healthcare for men and women, and empower the community to self-monitor and identify barriers to improving access to healthcare through the Community Engagement Core (CEC). The CEC aims to develop meaningful relationships among the research, medical, and lay communities promoting the increase of awareness of the importance of blood pressure control in women through community outreach, a Qualitative Needs Assessment, design and dissemination of unique illustrative educational materials related to cardiovascular disease in women, and the launch of a Blood Pressure Checks for Women campaign. In six months (December 2023 – May 2024), ROAR participated in 10 community events providing blood pressure checks and educational materials and reaching more than 600 people in 6 counties; awarded one pre-doctoral career enhancement mini-sabbatical; awarded three early career SABV pilot grants; hosted a SABV symposium reaching 120 biomedical scientists; and hosted a SABV seminar and workshop reaching 142 biomedical scientists. In conclusion, the ROAR CEC is well on its way to positively impacting hypertension perceptions and awareness in women across the Southeast.

Pressure‐induced constriction (PIC), also referred to as myogenic constriction, is an intrinsic response to the vascular smooth muscle cell (VSMC) of small arteries and arterioles. The response is mechano‐dependent and initiated by intraluminal pressure‐induced VSMC stretch. While numerous molecules are required for the overall signaling response, our laboratories have focused on the role of Degenerin (Deg) proteins, such as Epithelial Na+Channel (ENaC), because of their close evolutionary link to known mechanosensors in the nematode. Loss of function phenotypes, using broad spectrum pharmacological inhibition and specific gene silencing, have been demonstrated in cerebral arteries. However, a gain of function phenotype has not been shown. Demonstration of a gain of function phenotype is critical to establishing a role for loss of vascular degenerin function in certain diseases, such as pre‐eclampsia. Thus, the aim of this study was to determine if upregulation of full‐length βENaC in the isolated middle cerebral artery (MCA) enhances the PIC response. To address this aim, MCA segments were dissected from mature, female mice (24 weeks old, mixed genetic background) and transiently transfected with βENaC enhanced green fluorescent protein (EGFP) fusion vector (βENaC‐EGFP, n=5) or with EGFP alone (EGFP, n=5) using Lipofectamine 3000 (Thermo Fisher Scientific) using a 1:1 ratio. Vessel segments were incubated for 18 hours at 37 °C and 5% CO2, then pressurized in a pressure‐flow myograph system with physiological salt solution. Responses to KCl (20‐80 mM), phenylephrine (PE, 10‐7‐10‐4 M), and active (+Ca2+) and passive (‐Ca2+) diameters measured under graded pressure steps (15‐90 mm Hg, 15 mm Hg increments, 5 min) were determined to assess vasoconstriction to depolarizing and α‐adrenergic agonists and intraluminal pressure, respectively. Data were analyzed using repeated measures ANOVA. MCA vasoconstrictor responses to KCl (80 mM, 30.3±2.2% vs 34.3±4.2%, main effect of treatment p=0.55) and PE (10‐4 M, 40.3±4.0% vs 35.6.0±3.1%, main effect of treatment p=0.73) were identical in βENaC‐EGFP and EGFP transfected segments, respectively. Circumferential stress/strain, wall thickness and wall‐to‐lumen ratio were also similar. In contrast, PIC responses were greater in βENaC‐EGFP group at >30 mm Hg (main effect of treatment p=0.0015). Peak myogenic tone was nearly 2 fold greater in βENaC‐EGFP treated segments (11.6±0.4% vs 6.3±0.4% at 75 mmHg, p=0.0006). These data confirm previous findings that βENaC is an important mediator of the PIC response. Moreover, they provide proof‐of‐concept that upregulating βENaC can potentially improve PIC responses in cerebral vessels and will allow us to test the hypothesis that downregulation of vascular βENaC in inflammatory conditions, such as preeclampsia, contributes to cerebrovascular dysfunction.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder that predominately affects women and is associated with high rates of hypertension and cardiovascular disease. SLE is characterized by B and T lymphocyte dysfunction and altered cytokine production, including elevated levels of the adipocytokine leptin. Leptin has various immunomodulatory properties, including promoting the expansion of proinflammatory T lymphocytes as well as the proliferation and survival of B cells. Because B and T lymphocytes have been implicated in the development of autoimmune‐associated hypertension by our laboratory, we hypothesized that leptin antagonism would attenuate hypertension and renal injury in an experimental model of SLE. To test this hypothesis, 28 week old female SLE (NZBWF1, n=27) and control (NZW, n=25) mice were administered 3 mg/kg of murine leptin superantagonist (LA) or vehicle (0.2% NaHCO3, pH 8.0) every 48 hours for four weeks. At the conclusion of the study, immune cell populations in the peripheral blood were analyzed using flow cytometry. The percentages of CD45R+ B lymphocytes were not significantly different between vehicle and LA‐treated SLE mice (17.4±3.0% SLE‐vehicle vs. 11.7±1.9% SLE‐LA, p=0.1). Similarly, there were no significant changes in CD3+CD4+or CD3+CD8+T cells; however, double negative T cells (CD3+CD4−CD8−) were significantly decreased in SLE mice administered LA as compared to vehicle treated SLE mice (25.1±2.0% SLE‐vehicle vs. 16.4±2.9% SLE‐LA, p=0.03). Increased numbers of these proinflammatory T cells have been previously shown in both patients with SLE and in the NZBWF1 mouse. Mean arterial pressure (MAP; mmHg), measured in conscious mice by carotid artery catheter, was higher in SLE mice than in control mice (115±4 mmHg Controlvehicle vs. 131±2 mmHg SLE‐vehicle, p=0.01), and treatment with LA decreased blood pressure in SLE mice (SLE‐LA: 121±3 mmHg; p=0.03 vs. SLE‐vehicle). Urinary albumin excretion, a marker of renal injury, was assessed by ELISA at the conclusion of the study. Albumin excretion was increased in SLE mice (0.064±0.01 mg/day Control‐vehicle vs. 20.7±9.7 mg/day SLE‐vehicle, p=0.12), but urinary albumin remained elevated in SLE mice administered LA (16.4±6.8 mg/day, p=0.96 vs. SLE‐vehicle). An additional indicator of renal injury, B and T lymphocyte infiltration, was assessed by flow cytometry. SLE mice had significantly higher levels of renal CD45+B cells and CD3+T cells as compared to control mice, but SLE mice treated with LA had lower percentages of renal CD3+T cells as compared to SLE‐vehicle treated mice (2.7±0.6% SLE‐vehicle vs. 1.3±0.2% SLE‐LA, p=0.04). In conclusion, these data suggest that leptin plays a pathogenic role in the development of hypertension in SLE, in part by promoting the expansion of inflammatory T cells and the infiltration of T cells into the kidneys.Support or Funding InformationResearch supported by: VA Merit Award BX002604‐01A2 to MJR, NIH P01HL051971 and P20GM104357 to UMMC‐Department of Physiology and Biophysics, and NIH NHLBI F32HL137393 to EBT.

Systemic lupus erythematosus (SLE) is a chronic multi‐system autoimmune disease characterized by the presence of circulating autoantibodies, inflammation, hypertension, renal injury, and cardiovascular disease. Current immunosuppressive therapies often impose unwanted side effects leaving patients searching for alternative therapeutic options. Curcumin has been used in Eastern medicine for its natural anti‐inflammatory and antioxidant properties. We hypothesize that the administration of curcumin will lessen renal inflammation and injury in female mice with SLE.Female NZBWF1 mice, an established experimental model of SLE, and female NZW (control) mice were administered curcumin (500 mg/kg/day) or corn oil (vehicle) by oral gavage for 14 days. This design is based on previously published experimental data using curcumin in mice. Plasma blood urea nitrogen (BUN), a clinical marker of renal function, was assessed at baseline (26 weeks of age) and again at the end of the study (28 weeks of age). To assess immune activity, we measured circulating anti‐dsDNA IgG autoantibodies, spleen weight, and used flow cytometric analyses of peripheral blood leukocytes to assess circulating B lymphocytes at the same time points.Compared with vehicle treated SLE mice, curcumin treated mice with SLE have significantly lower BUN at 28 weeks of age (13.7 ± 0.6 mg/dL SLE‐curcumin, n=14 vs. 36.2 ± 11.4 mg/dL SLE‐vehicle, n=5) (P = 0.0030). As expected, spleen weights are increased in female mice with SLE. However, curcumin treated SLE mice at 28 weeks of age have lower spleen weights compared to SLE vehicle treated mice (0.114 ± 0.009 g SLE‐curcumin, n=14 vs. 0.212 ± 0.03 g SLE‐vehicle, n=5) (P = 0.0008). In addition, the percentage of circulating CD45R+ B cells are decreased at 28 weeks of age in SLE curcumin treated mice compared to vehicle treated SLE mice (5.6 ± 1.6% SLE‐curcumin, n=4 vs. 36.4 ± 9.5% SLE‐vehicle, n=10) (P = 0.068). Despite fewer circulating CD45R+ B cells, plasma levels of anti‐dsDNA IgG autoantibodies are unchanged in curcumin treated mice with SLE compared to vehicle treated mice with SLE. These data suggest that curcumin modulates autoimmune activity and may lessen renal injury in female mice with SLE.Support or Funding InformationResearch reported in this publication was supported by the NIGMS of the NIH (5U54GM115428 and P20GM104357), NHLBI of the NIH (P01HL051971), VA Merit (BX002604‐01A2), and AHA pre‐doctoral fellowship (19PRE34380830).

Cerebrovascular events account for ~40% of all preeclamptic/eclamptic‐related deaths and can induce neurological complications. While placental ischemia can impair cerebrovascular function in women with preeclampsia (PE) and rats which develop a PE‐like syndrome, the placental‐derived factors mediating these effects are elusive. Although pro‐inflammatory cytokines, including Interleukin‐17 (IL‐17), are increased by placental ischemia, it is not known whether IL‐17 disrupts cerebrovascular function during pregnancy. Therefore, we tested the hypothesis that infusion of IL‐17 during pregnancy impairs cerebral blood flow (CBF) autoregulation and myogenic tone, while also promoting increased regional blood‐brain‐barrier (BBB) permeability and brain water content (BWC). Pregnant Sprague Dawley rats were infused with recombinant IL‐17 (150 pg/day) via mini‐osmotic pump (i.p.) from gestational days 14–19. Cerebral blood flow was measured by laser Doppler flowmetry and myogenic tone was assessed in isolated middle cerebral arteries (MCA) by pressure‐flow myography. BWC was determined by quantifying dry:wet weight ratio and BBB permeability was measured by Evans blue extravasation. As previously shown, IL‐17 increased MAP (112 ± 2.5 vs 99.7 ± 2.8 mmHg; p<0.01) compared to pregnant control rats. CBF was significantly greater in IL‐17‐treated pregnant rats at 180 mmHg (187 ± 19 vs 149 ± 22%; p<0.05) and 190 mmHg (215 ± 18 vs 160 ± 23%; p<0.01) compared to pregnant control rats. Myogenic tone peaked at 125 mmHg in the MCA of IL‐17‐treated rats and declined thereafter, forming an inverted‐U curve in response to increases in intraluminal pressure. In contrast, myogenic tone peaked at 175 mmHg in control pregnant rats, forming a hyperbolic curve, which suggests that myogenic responses are blunted at high arterial pressures in IL‐17‐treated rats. BWC tended to increase in the cortex (80.5 ± 0.2 vs. 79.5 ± 0.4%, p = 0.054) and striatum (77.6 ± 0.8 vs. 75.1 ± 1.4%, p = 0.062), and was significantly increased in the posterior cerebrum (79.7 ± 0.4 vs. 77.8 ± 0.5%, p< 0.01) and cerebrum (79.8 ± 0.3 vs. 78.8 ± 0.1%, p< 0.01) of IL‐17‐treated rats. IL‐17 increased BBB permeability in the cortex only (0.017 ± 0.002 vs 0.012 ± 0.002, p<0.05). These data suggest that increases in circulating IL‐17 during pregnancy could contribute to cerebrovascular dysfunction leading to impaired CBF control and myogenic responses, cerebral edema, and increased BBB permeability. IL‐17 may be a potential therapeutic target to prevent or ameliorate neurological impairment in preeclampsia. Future studies examining whether blockade of IL‐17 in placental ischemic rats could prevent cerebrovascular dysfunction may provide valuable insight into the role of cytokines on cerebrovascular health during pregnancy.Support or Funding InformationFunding: R01HL12186106, R01HL136684‐02, R01HL136684, P01HL051971, P20GM104357, 5U54GM115428, T32HL105324, and 19POST34450074.

Systemic lupus erythematosus (SLE) is a chronic multi‐system autoimmune disease characterized by the presence of circulating autoantibodies, systemic and localized inflammation, prevalent hypertension, renal injury, and cardiovascular disease. Onset of the disease typically occurs in young women of child‐bearing age, a time when women are typically protected from cardiovascular risk factors like hypertension. Although kidney involvement is common to patients with SLE, and renal hemodynamic dysfunction is evident in patients with active renal disease, surprisingly little is known about early changes in renal hemodynamic function that may contribute to the pathogenesis of hypertension during autoimmune diseases. We hypothesize that the loss of immunological tolerance and subsequent production of autoantibodies in SLE leads to impaired renal hemodynamic function that precedes the development hypertension. Female NZBWF1 mice, an established experimental model of SLE, and female NZW (control) mice were instrumented with carotid artery and jugular vein catheters to determine mean arterial pressure (MAP) and glomerular filtration rate (GFR) respectively at ages 15, 20, 28, 31, and 34 weeks (n>3 control/age, n>4 SLE/age). MAP was measured in conscious, freely‐moving mice. GFR was measured by the clearance of fluorescein isothiocyanate‐inulin (FITC‐inulin) after achieving steady state through continuous infusion. From 15 weeks of age to 34 weeks of age, there is a progressive increase in the levels of circulating autoantibodies in female mice with SLE, whereas autoantibodies are unchanged in control mice (Figure 1). Compared with control mice, female mice with SLE have higher MAP at 15, 20, 28, 31, and 34 weeks of age, with an increase in pressure that occurs by 34 weeks of age (Figure 2). GFR remains moderately higher in SLE mice compared with control mice until 28 weeks of age at which time GFR sharply declines in the mice with SLE (Figure 3). GFR is unchanged in control mice across all ages. These data suggest that changes in renal hemodynamic function occur in female SLE mice prior to changes in MAP suggesting an important mechanistic role for autoimmunity to directly impair renal hemodynamic function and promote the development of hypertension. Support or Funding Information VA Merit BX002604, American Heart Association Predoctoral 19PRE34380830, NIH P20GM104357, NIH 5U54GM115428, NIH 5P01HL051971 This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Systemic lupus erythematosus (SLE) is a chronic multisystem autoimmune disorder that predominately affects women of childbearing age and is associated with high rates of hypertension, renal injury, and cardiovascular disease. SLE disease is characterized by B and T lymphocyte dysfunction, leading to the production of pathogenic autoantibodies to a variety of self components. Autoantibodies produced during SLE form immune complexes that deposit in endothelial basement membranes and contribute to vascular inflammation. Autoantibodies also directly bind to endothelial cells in vitro in an antigen‐specific manner. However, whether vascular function is directly affected by autoantibodies in SLE is unknown. Our laboratory previously reported that an established female mouse model of SLE (NZBWF1) progressively develops impaired vascular relaxation when compared to age matched healthy controls (female NZW). We hypothesize that the depletion of plasma cells, which are responsible for the majority of immunoglobulin production, with the proteasome inhibitor bortezomib will improve endothelial function in NZBWF1 mice. To test this, 12 week old control (n=14) and SLE (n=12) mice were injected i.v. with vehicle (0.9% NaCl) or bortezomib (0.75 mg/kg) twice weekly for 12 weeks. Treatment with bortezomib lowered the percentages of CD138 + plasma cells in the spleen of SLE animals, as assessed by flow cytometry (0.51±0.21 vs. 0.26±0.06%, p=0.06). In addition, total circulating IgG concentrations were determined using ELISA. Plasma IgG levels were higher in SLE mice as compared to control animals (2.0±0.15 vs. 6.1±1.0 mg/mL, p<0.01), and bortezomib treatment significantly lowered IgG levels in both SLE and control animals (SLE‐ 6.1±1.0 vs. 2.3±0.46 mg/mL, p<0.001; Control‐2.0±0.15 vs. 1.1±0.09 mg/mL, p<0.001). Circulating anti‐dsDNA autoantibodies, characteristic of human SLE, were also measured. Anti‐dsDNA IgG was higher in SLE mice as compared to controls (13.1±2.2 kU/mL vs. 522±291 kU/mL, p=0.09). Bortezomib‐treated SLE mice had lower levels of anti‐dsDNA IgG (522±291 vs. 12.0±2.7 kU/mL, p=0.08). To assess the impact of antibody depletion on vascular function, carotid rings were suspended in organ chamber baths and endothelium‐dependent and endothelium‐independent concentration responses (10 −8 to10 −4 M) to acetylcholine (ACh) and sodium nitroprusside (SNP) were assessed in vessels pre‐contracted with the thromboxane mimetic U46619 (0.4 μg/mL). ACh‐mediated relaxation was impaired in SLE‐vehicle mice as previously reported by our laboratory; however, relaxation was improved in bortezomib‐treated SLE mice compared to vehicle‐treated SLE mice (Figure; *, p<0.05 vs. all other groups; #, p<0.05 SLE‐vehicle vs. control groups and SLE‐bortezomib vs. control groups). Bortezomib treatment did not significantly alter SNP‐mediated relaxation in any treatment group. Taken together, these data suggest that autoantibodies play an important mechanistic role in the development of endothelial‐dependent vascular dysfunction. Support or Funding Information Research supported by: VA Merit Award BX002604‐01A2 to MJR and NIH NHLBI PO1HL051971 and P20GM104357 to UMMC‐Department of Physiology and Biophysics. EBT is supported by NIH NHLBI F32HL137393. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by inflammation, hypertension, renal injury, and cardiovascular disease. Kidney involvement is common to patients with SLE and renal hemodynamic dysfunction is evident in patients with active renal disease. However, little is known about early changes in renal hemodynamic function that may contribute to the pathogenesis of hypertension during SLE. We hypothesize that the loss of immunological tolerance and subsequent production of autoantibodies in SLE leads to impaired renal hemodynamic function that precedes the development hypertension. Female NZBWF1 (SLE) and female NZW (control) mice were instrumented with carotid artery and jugular vein catheters to determine MAP and glomerular filtration rate (GFR) respectively at ages 15, 20, 24, 28, 31, and 34 weeks. MAP was measured in conscious, freely-moving mice. GFR was measured by the clearance of fluorescein isothiocyanate-inulin (FITC-inulin) after achieving steady state through continuous infusion for five hours. GFR increases at 28 weeks of age followed by a significant decline by 34 weeks of age (P = 0.0127, P = 0.0001) compared to 34 and 15 weeks of age respectively, in the mice with SLE. GFR is increased in control mice by 28 weeks of age (P = 0.002) and remains unchanged at other time points. Compared with control mice, female mice with SLE have higher MAP by 34 weeks of age (P = <0.0001). These data suggest that changes in renal hemodynamic function occur in female SLE mice prior to changes in MAP suggesting an important mechanistic role for autoimmunity to directly impair renal hemodynamic function and promote the development of hypertension.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder that predominately affects women and is associated with high rates of hypertension and cardiovascular disease. SLE is characterized by B and T lymphocyte dysfunction and altered cytokine production, including elevated levels of the adipocytokine leptin. Leptin promotes the expansion of proinflammatory T helper (T H ) 1 and T H 17 cells and inhibits the differentiation of regulatory T cells (T REG ). Because T cells are mechanistically linked to the development of hypertension in SLE, we hypothesized that administration of leptin would alter T H subsets and accelerate the development of hypertension in a female mouse model of SLE. To test this hypothesis, 30 week old female SLE (NZBWF1, n=21) and control (NZW, n=25) mice were implanted with minipumps to continuously deliver recombinant mouse leptin (0.5 mg/kg/day) or vehicle for four weeks. At the conclusion of the study, splenic CD4 + T H subsets were analyzed by flow cytometric analyses and no significant alterations in T H 1, T H 2, T H 17, or T REG cells were detected; however, circulating T REG in SLE mice were lower after leptin administration (1.1±0.2% SLE-vehicle vs. 0.52±0.1% SLE-leptin, p<0.05). Circulating IgG isotypes were also analyzed to correlate with T H subset activities and circulating levels of IgG2a were higher in SLE mice administered leptin (0.79±0.2 mg/mL SLE-vehicle vs. 1.2±0.05 mg/mL SLE-leptin, p<0.05), indicating increased T H 1 activity. In further support of inflammatory T cell activity, circulating levels of TNF-α and IL-17 were elevated in SLE mice administered leptin (TNF-α: 18.6±8.4 pg/mL SLE-vehicle vs. 80.5±15.9 pg/mL SLE-leptin, p<0.01) (IL-17: 0.46±0.1 pg/mL SLE-vehicle vs. 1.68±0.5 pg/mL, p<0.01). Mean arterial pressure (MAP; mmHg), measured in conscious mice by carotid catheter, was higher in SLE mice than in control mice (113±3 Control-vehicle vs. 128±3, p=0.06), and leptin further increased blood pressure in SLE mice (SLE-leptin: 134±3; p<0.01 vs. Control-vehicle and Control-leptin). In conclusion, these data suggest that leptin plays a pathogenic role in the development of hypertension in autoimmunity, in part by altering the balance of T H 1, T H 17, and T REG activity.

Preeclampsia is a multi‐system disorder of pregnancy characterized by widespread vascular dysfunction. Though the precise etiology of preeclampsia remains largely enigmatic, emerging evidence suggests that an ischemic placenta releases circulating vasoactive factors all of which culminate in endothelial dysfunction. The incretin hormone glucagon‐like peptide 1 (GLP‐1) has been extensively studied for its therapeutic benefit in diabetes mellitus, where it acts to enhance endogenous insulin secretion. In addition to its role in glucose homeostasis, GLP‐1 receptors are present on endothelial cells and activate the endothelial nitric oxide synthase (eNOS) signaling axis. In turn, GLP‐1 receptor agonists have been demonstrated to improve endothelial dysfunction. Therefore, we hypothesized that administration of a GLP‐1 receptor agonist would attenuate endothelial dysfunction and improve blood pressure in our model of preeclampsia.In order to test this hypothesis, timed pregnant CD‐001 rats (an outbred sub‐strain of Sprague‐Dawley rats) underwent surgery on gestation day 15 to place silver clips on the aorta and ovarian‐uterine arterial anastomoses. This procedure restricts blood flow to the fetal‐placental unit by 60%, producing a model of reduced uterine perfusion pressure (RUPP) that mimics several characteristics of human preeclampsia. From gestation day 15 to 20, animals received daily subcutaneous injections of 0.3 mg/kg liraglutide or vehicle. Animals were instrumented with carotid catheters on gestation day 19 and blood pressure determined on day 20 while the animals were conscious and restrained. Daily weight, food intake, and pup outcome were collected.We found that blood pressure was significantly lower in RUPP animals treated with liraglutide (107.8 ±2.5 vs 117.6 ±4.0 mmHg, n = 5 and 8 respectively, p<0.05). Though food intake was diminished in the liraglutide‐treated group immediately following surgery (2.0 ±0.6 vs. 10.8 ±1.6 grams/day on gestation day 15), it rapidly recovered (16.3 ±1.6 vs. 14.5 ±1.8 grams/day on gestation day 19), and there was no significant difference between body weight (259±10 vs 287 ±14 grams on gestation day 20), litter size (8 ±2 vs 10 ±1 live pups), or pup weight (2.07 ±0.05 vs 2.15 ±0.11 grams).This data shows that GLP‐1 receptor agonism effectively lowers blood pressure in a placental ischemic model of preeclampsia. Although the mechanisms contributing to the improved blood pressure remain unclear, these data suggest that the GLP‐1 system represents a new potential therapeutic target in preeclampsia. Importantly, given its role in diabetes, several GLP‐1 receptor agonists are already FDA approved. Future studies will examine vascular mechanisms by which GLP‐1 receptor agonism protects against the development of placental ischemia‐induced hypertension.Support or Funding Information18PRE34050024HL136684P20GM104357This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Systemic lupus erythematosus (SLE) is a prototypic multisystem autoimmune disorder that is characterized by a loss of immunological tolerance and the expansion of autoreactive T and B lymphocytes, leading to the production of autoantibodies. The autoantibody production leads to downstream chronic inflammation resulting in high rates of hypertension, renal injury, and cardiovascular disease in patients with SLE. The immunomodulatory adipocytokine leptin plays a key role in the maintenance and development of inflammation, in part by promoting the expansion of proinflammatory helper T cells and inhibiting the differentiation of regulatory T cells (TREG). Circulating levels of leptin are elevated in patients with SLE, but it is unclear whether leptin plays a direct role in the pathogenesis of SLE. In the present study, we hypothesized that administration of leptin will accelerate the progression of disease in a female mouse model of SLE. To test this hypothesis, 30 week old female SLE (NZBWF1, n=10) and control (NZW, n=14) mice were implanted with microosmotic pumps to continuously deliver recombinant mouse leptin at a rate of 0.5 mg/kg/day or vehicle (0.9% NaCl) for four weeks. Plasma leptin levels, as measured by ELISA, were significantly increased in both control (10.6±1.6 ng/mL Control‐vehicle vs. 29.5±5.4 ng/mL Control‐leptin, p<0.01) and SLE mice who received leptin (21.0±6.4 ng/mL SLE‐vehicle vs. 50.3±17.5 ng/mL SLE‐leptin, p<0.05). Leptin administration significantly decreased food intake in SLE mice (4.0±0.15 g/day SLE‐vehicle vs. 3.2±0.10 g/day SLE‐leptin, p<0.05), but did not alter food intake in control mice (3.7±0.16 g/day Control‐vehicle vs. 3.3±0.25 g/day Control‐leptin, p=0.33). Body composition was assessed using Echo MRI and fat mass, as a percentage of body weight, was not changed in control mice (19.8±1.3% Control‐vehicle vs. 20.8±1.6% Control‐leptin p=0.99), but was significantly lower in SLE‐leptin treated mice (24.7±2.4% SLE‐vehicle vs. 12.2±4.8% SLE‐leptin, p<0.05). Circulating levels of anti‐dsDNA autoantibodies, a marker of SLE disease activity, were higher in SLE mice compared to controls (0.56±0.16 Control‐vehicle vs. 0.98±0.16 SLE‐vehicle OD450, p<0.05), and these levels were further increased in leptin infused SLE mice compared to vehicle‐infused controls (0.98±0.16 SLE‐vehicle vs. 1.3±0.12 SLE‐leptin OD450, p<0.05). Circulating CD4+FoxP3+ TREG, as assessed by flow cytometry, were lower in SLE mice than in control mice (2.88±0.44% Control‐vehicle vs. 1.44±0.16% SLE‐vehicle, p<0.05), as previously reported by our laboratory. Leptin administration decreased the levels of circulating TREG in both control and SLE animals. Leptin infusion also increased the prevalence of mice with albuminuria (60% of SLE mice administered leptin vs. 25% of vehicle‐treated mice), as measured by dipstick assay. Taken together, these data suggest that SLE mice may have enhanced sensitivity to leptin and that increased leptin levels exacerbate disease activity. In addition, therapeutics aimed at modulating leptin activity could have potential benefit for patients with SLE.Support or Funding InformationResearch supported by: VA merit award # I01BX002604‐01A2 to MJR and NHLBI F32HL137393 to EBT.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Cerebral complications are a leading cause of morbidity and mortality in preeclamptic pregnancies, accounting for approximately 40% of all preeclampsia/eclampsia related deaths. However, the underlying pathophysiological mechanisms remain unclear. Given the central role of the ischemic placenta in driving preeclampsia, we hypothesized that circulating factors released from the ischemic placenta directly impair cerebral vascular function and blood brain barrier permeability. Specifically, the role of TNFα, an inflammatory cytokine increased in the placenta and circulation of preeclamptic pregnancies, was examined. Pregnant Sprague Dawley Rats were infused with vehicle or TNFα (200 ng/day i.p, from gestational day 14 to 19). Evan’s blue extravasation was measured in the cerebral cortex as an indicator of blood brain barrier (BBB) permeability, and brain water content was assessed as an indicator of cerebral edema at the time of euthanasia. In a separate experimental group, rats were infused with vehicle or TNFα in order to examine the impact on cerebral blood flow (CBF) autoregulation during pregnancy. CBF was measured by laser Doppler flowmetry under anesthesia. Rats were intubated and ventilated in order to maintain constant blood gases, and mean arterial pressure was elevated step-wise from 100-190 mmHg by infusion of phenylephrine to determine changes in CBF. Infusion of TNFα in pregnant rats caused a significant 10 mmHg increase in arterial pressure (n = 12, p < 0.05). Brain water content was increased in TNFα infused rats (p < 0.05), mainly in the anterior cerebrum, and significantly increased BBB permeability in the hippocampus (p < 0.05). TNFα infusion impaired CBF autoregulation in pregnant rats with increased CBF flow at 180 mmHg (253 ± 16 vs 188 ± 9, n = 6, p < 0.05). Peak carbon dioxide was not different between groups. These data suggest that increasing TNFα to levels observed during preeclampsia results in cerebral vascular changes that may mechanistically underlie the increased risk for encephalopathy.

Primary aldosteronism is characterized by excess aldosterone secretion by the adrenal gland independent of the renin-angiotensin system and accounts for ~10% of hypertensive patients. Excess aldosterone causes cardiac hypertrophy, fibrosis, inflammation, and hypertension. The molecular mechanisms that trigger the onset and progression of aldosterone-mediated cardiac injury remain incompletely understood. MicroRNAs (miRNAs) are endogenous, small, noncoding RNAs that have been implicated in multiple cardiac pathologies; however, their regulation and role in aldosterone-mediated cardiac injury and dysfunction remains mostly unknown. We previously reported that microRNA-21 (miR-21) is the most upregulated miRNA by excess aldosterone in the left ventricle in a rat experimental model of primary aldosteronism. To elucidate the role of miR-21 in aldosterone-mediated cardiac injury and dysfunction, miR-21 knockout mice and their wild-type littermates were treated with aldosterone infusion and salt in the drinking water for 2 or 8 wk. miR-21 genetic ablation exacerbated aldosterone/salt-mediated cardiac hypertrophy and cardiomyocyte cross-sectional area. Furthermore, miR-21 genetic ablation increased the cardiac expression of fibrosis and inflammation markers and fetal gene program. miR-21 genetic ablation increased aldosterone/salt-mediated cardiac dysfunction but did not affect aldosterone/salt-mediated hypertension. miR-21 target gene Sprouty 2 may be implicated in the cardiac effects of miR-21 genetic ablation. Our study shows that miR-21 genetic ablation exacerbates aldosterone/salt-mediated cardiac hypertrophy, injury, and dysfunction blood pressure independently. These results suggest that miR-21 plays a protective role in the cardiac pathology triggered by excess aldosterone. Furthermore, miR-21 supplementation may be a novel therapeutic approach to abolish or mitigate excess aldosterone-mediated cardiovascular deleterious effects in primary aldosteronism.

Preeclampsia is clinically defined as elevated blood pressure after 20 weeks gestation along with other major organ involvement, such as brain, renal, or liver. It impacts 3–8 % of pregnancies, and there is currently no cure other than delivery of the placenta. The mechanisms leading to preeclampsia are not fully understood; however, recent work by our laboratory showed that the pregnant Dahl S rat closely mimics women with preexisting hypertension that develop preeclampsia. Therefore, pregnant Dahl S rats can be utilized to better understand the pathophysiological mechanisms and consequences of superimposed preeclampsia. Impaired cerebral vascular function has been implicated as a potential mechanism contributing to the increased risk of neurological complications associated with preeclampsia. In this study, we tested the hypothesis that pregnancy induces vascular permeability in cerebral arteries in Dahl S rats. Female Dahl S rats were mated at 16–18 weeks of age, while age‐matched virgin littermates were used as controls (n=5 per group). Middle cerebral arteries were isolated on gestational day 19–21, cannulated with glass pipettes, and pressurized to assess arterial permeability by measuring the change in intravascular pressure over time. Starling's Law states that fluid flux across a vessel wall is the result of hydraulic pressures and oncotic pressures due to differences in osmolality. In our model, the oncotic pressure gradient across the vessel wall was 0 mmHg, leaving only hydraulic pressures to determine fluid flux; therefore, the rate in pressure drop measured inside an isolated vessel is an indirect assessment of vascular permeability. Isolated vessels were equilibrated at 37°C for 30 minutes in physiological salt solution and pressurized to 75 mmHg. The diameter of isolated arteries after equilibration was 162±11 μm and 157±11 μm in virgin and pregnant rats, respectively (p=0.51). Intraluminal pressure was recorded every 5 minutes for one hour. As shown in the figure, the drop in intravascular pressure is greater in vessels isolated from pregnant rats compared to the pressure drop in vessels from virgin rats (*p< 0.01 vs virgin, n=5). Over the 60 minute experimental period, there was a significant difference in the drop in pressure permeability between the virgin and pregnant Dahl S rats (24.4±5.0 mmHg vs. 50.2±3.7 mmHg, respectively, P<0.01, n=5). These data indicate that pregnancy increases cerebral arterial permeability in Dahl S rats.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Humans have had a long and complex relationship with salt. Although highly valued in many societies, dietary salt has long been associated with high blood pressure1–3 and, more recently, with other diseases.4–6 Some individuals with hypertension often display salt-sensitive blood pressure changes, which is a condition more prevalent among blacks, older people, and individuals with renal insufficiency or diabetes mellitus.7–9 In general, for those with salt-sensitive hypertension, excess sodium intake is associated with higher blood pressure, whereas a low-salt diet decreases blood pressure.3 In spite of this well-known association, the basic molecular and cellular mechanisms underlying the effects of salt on blood pressure regulation are still not well understood. Furthermore, individuals with high blood pressure are at increased risk for multiple diseases (ie, coronary artery disease, heart failure, stroke, and renal disease) although at present whether or not a high dietary salt intake can directly lead to these diseases (ie, in the absence of hypertension) is not known. Our understanding of the effect of salt on health has grown even more complex recently. Researchers have reported a new connection between salt and autoimmunity: a high-salt diet was shown to accelerate autoimmune activity in a mouse model of multiple sclerosis.10,11 In addition, a close connection between hypertension and the immune system has been revealed.12–16 However, the causal relationships between salt, immunity, and hypertension (eg, how salt could mediate interactions between the immune system and the vasculature, brain, or kidney to increase blood pressure) are not well understood. The National Heart, Lung, and Blood Institute convened a Working Group (WG) in 2014 to discuss this new emerging scientific area in hypertension research. The WG brought together experts from diverse backgrounds including hypertension, epidemiology, preeclampsia, cardiovascular disease, …

Mycophenolate mofetil (MMF) is an immunosuppressive agent that selectively depletes proliferating lymphocytes and is commonly used to prevent organ transplant rejection or treat autoimmune disease. Systemic lupus erythematosus (SLE) is a chronic multisystem autoimmune disorder that predominately affects women of childbearing age and is associated with a high rate of hypertension, renal injury, and cardiovascular disease. Recent clinical evidence suggests that treatment with MMF can reduce blood pressure in patients with rheumatoid arthritis or psoriasis; however, whether MMF can be effectively utilized in the control of blood pressure during SLE remains unclear. Because immune dysregulation and inflammation have been linked to the pathophysiology of essential hypertension, we hypothesized that MMF will attenuate hypertension and renal injury in an experimental mouse model of SLE. Female SLE (NZBWF1) and control (NZW) mice, aged 26 weeks, were injected intraperitoneally with 5% dextrose (vehicle) or MMF daily at a rate of 60 mg/kg/day for eight weeks. CD45R + B cells were lower in the peripheral blood of SLE mice treated with MMF for eight weeks (19.5±1.3% SLE‐Vehicle vs. 5.2±1.5% SLE‐MMF, p< 0.01), but the levels of both CD4 + and CD8 + T cells were unchanged. Circulating levels of anti‐dsDNA IgG auto antibodies, a marker of SLE disease activity, were higher in SLE mice as compared to control mice (789±120 vs. 161±57 units/mL, p<0.0001), and were significantly lower in mice treated for eight weeks with MMF (SLE‐MMF: 321±90; p<0.05 vs. SLE‐Vehicle). Urinary albumin excretion, an indicator of glomerular injury, was increased in SLE mice as compared to controls (5.6±1.9 vs. 0.11±0.02 ng/day, p<0.0001), and was lower in SLE mice treated with MMF (SLE‐MMF: 0.38±0.20 ng/day; p<0.001 vs. SLE‐Vehicle). Mean arterial pressure (MAP; mmHg) measured in conscious mice by carotid catheter was higher in SLE mice than in control mice (141±3 vs. 120±3, p<0.05). MAP was significantly lower in SLE mice treated with MMF when compared to vehicle treated SLE mice (SLE‐MMF: 120±5; p<0.001 vs. SLE‐Vehicle). These data suggest that MMF treatment may have added clinical benefit for patients with SLE by helping to control blood pressure, a major contributor to cardiovascular risk in this population. Moreover, given that MMF selectively depleted CD45R + B cells, these data provide additional evidence that B cells and the production of auto antibodies mechanistically contribute to the pathogenesis of hypertension during SLE. Support or Funding Information Research supported by: VA# I01BX002604‐01A2 (MJR), T32HL105324 (EBT), HL051971 (UMMC‐Physiology)

Cerebrovascular events contribute to ~40% of preeclampsia/eclampsia related deaths, and neurological symptoms are common among preeclamptic patients. We previously reported that placental ischemia, induced by reducing utero‐placental perfusion pressure, leads to impaired cerebrovascular myogenic reactivity and cerebral edema in the pregnant rat. Whether the impaired myogenic reactivity is associated with altered cerebral blood flow (CBF) autoregulation and the edema is due to altered blood‐brain barrier (BBB) permeability is unclear. Therefore, we tested the hypothesis that placental ischemia leads to impaired CBF autoregulation and a disruption of the BBB. CBF autoregulation, measured in vivo by laser Doppler flowmetry, was significantly impaired in placental ischemic rats compared to normal pregnant rats. Brain water content was increased in the anterior cerebrum of placental ischemic rats and BBB permeability, assayed using the Evans blue extravasation method, was also increased in the anterior cerebrum. The expression of the tight junction proteins: claudin‐1 was increased in the posterior cerebrum while zonula occludens‐1, and occludin, were not significantly altered in either the anterior or posterior cerebrum. These results are consistent with the hypothesis that placental ischemia mediates anterior cerebral edema through impaired CBF autoregulation and the associated increased transmission of pressure to microvessels, resulting in increased BBB permeability and cerebral edema.

Systemic lupus erythematosus (SLE) is an autoimmune disorder with prevalent renal disease, for reasons that continue to be elucidated. Recent evidence suggests that dietary salt may be an important environmental factor that promotes some autoimmune diseases. Therefore, we hypothesized that a long‐term high salt diet would accelerate the progression of renal disease during SLE. In order to test this, an established experimental model of SLE (female NZBWF1 mice) was fed a standard (0.4% NaCl) or high salt (4% NaCl) diet starting at 10 weeks of age. Sodium intake (in meq/day) was measured three times throughout the study and was significantly greater in high salt fed animals at 16 weeks of age (2.9±0.6 vs. 0.5±0.06 , p=0.01), 23 weeks (4.8±0.8 vs. 0.6±0.06, p=0.004) and 29 weeks (5.7±0.6 vs. 0.6±0.05, p=0.0007). Urinary albumin was monitored monthly until 30 weeks then weekly by dipstick assay as a marker of renal injury until 34 weeks of age at which time the mice were euthanized and the kidneys were harvested. Renal cortex enriched mRNA was analyzed by qRT‐PCR for inflammatory markers. 80% of animals (n=5) on a normal diet developed albuminuria (蠅 100 mg/dL), whereas none (0%, n=5) of the high salt fed animals developed albuminuria. Renal cortical interleukin‐2 (IL‐2), a cytokine important for T cell differentiation, expression was significantly lower in high salt fed animals (0.26±0.14 relative to normal chow fed mice, p=0.03). These data suggest that, contrary to the original hypothesis, a long‐term high salt diet may protect against the renal injury associated with SLE, possibly through alterations in IL‐2 mediated T cell differentiation. Furthermore, the data advance our overall understanding of how dietary salt may impact different autoimmune disorders.Grant Funding Source: Supported by T32HL105324, 12GRNT12060203, HL085907, and HL051971

Despite preeclampsia being one of the leading causes of maternal death and a major contributor of maternal and perinatal morbidity, the mechanisms responsible for its pathogenesis have yet to be fully elucidated. Growing evidence indicates that reduced uteroplacental perfusion and the resulting placental ischemia triggers the cascade of events leading to this maternal disorder. While the well-established rat model of reduced uterine perfusion pressure (RUPP) is providing invaluable insight into the etiology of preeclampsia, the aim of this study was to develop a mouse model of reduced uterine perfusion to expand mechanistic investigation by incorporation with novel gene-targeted mice. To accomplish this aim, a sham surgical procedure or a restriction of blood flow at the abdominal aorta and the ovarian arteries was initiated at day 13 of gestation in C57BL/6J mice. Mean arterial pressure measured in conscious, chronically instrumented mice was significantly elevated in the RUPP (120 ± 4 mmHg) compared with the sham (104 ± 4 mmHg) mice at day 18 of gestation ( P < 0.01). Placental ischemia reduced fetal weights (0.95 ± 0.04 and 0.80 ± 0.02 g; RUPP vs. Sham, respectively; P < 0.02) and increased circulating levels of antiangiogenic soluble fms-related tyrosine kinases (sFlt)-1 ( P < 0.05) in the RUPP at day 18 of gestation. Plasma concentrations of sFlt-1 are increased in preeclamptic patients and in response to reduced uterine perfusion in the rat. Thus, these results suggest that the mouse model of reduced uterine perfusion is applicable to facilitate novel mechanistic investigation into the etiology of hypertension that results from placental ischemia during pregnancy.