bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 051: High Salt Enhances ROS and Ang II Contractions of Glomerular Afferent Arterioles From Mice With Reduced Renal Mass
Authors: Li Lingli , Lai EnYin , Luo Zaiming , Solis Glenn , Mendonca Margarida , Griendling Kathy , Wellstein Anton , Welch William , WILCOX Christopher S
Published: December 2018

High salt intake, angiotensin II (Ang II), and reactive oxygen species (ROS) enhance progression of chronic kidney disease (CKD). We reported that myogenic contractions of renal afferent arterioles (RAAs) were enhanced by superoxide (O
2
·-
) generated from p47
phox
/NOX2 but inhibited by H
2
O
2
generated from POLDIP2/NOX4. We tested the hypothesis that feeding a high salt diet to mice with the reduced renal mass (RRM) model of CKD generates specific ROS in their RAAs that enhances Ang II contractions.

Methods and Results:
C57BL/6 mice received surgical RRM or sham operations and 6% or 0.4% NaCl salt for 3 months. Ang II contractions were measured in RAAs perfused at 45 mmHg and superoxide (O
2
·-
) and H
2
O
2
by fluorescence microscopy. RRM enhanced the gene expression in RAAs for p47
phox
and NOX2 and high salt intake in mice with RRM enhanced the gene expression for AT1Rs, POLDIP2 and NOX4 and reduced the gene expression for catalase. Mice with RRM fed a normal salt diet had contractions to 10
-6
mol·l
-1
Ang II similar to sham (-56 ± 5 vs -52 ± 5 %; NS). However, RRM mice fed a high salt diet had an enhanced O
2
·-
and H
2
O
2
generation (P<0.005) with Ang II in RAAs and enhanced Ang II maximal contractions (-72 ± 2 vs -45 ± 2%; P<0.005) that were dependent on O
2
·-
from NOX2 since they were prevented in p47
phox
-/- mice and on H
2
O
2
from NOX4 since they were prevented in mice with transgenic smooth muscle cell expression of catalase (tg
CAT-SMC
), and in POLDIP2 +/- mice. However, RAA contractions to lower concentrations of Ang II (10
-8
to 10
-11
mol·l
-1
) were paradoxically enhanced in tg
CAT-SMC
vs Wt mice (-17 ± 2 vs -1 ± 1%; P<0.01) and in POLDIP2 +/- vs +/+ mice (-22 ± 3 vs -5 ± 3; P<0.01). Tempol normalized the ROS and Ang II contractions in RAAs from mice with RRM. In conclusion, both O
2
·-
from p47
phox
/NOX2 and H
2
O
2
from NOX4/POLDIP2 enhance maximal Ang II contractions of RAAs from mice with RRM fed a high salt diet but H
2
O
2
from NOX4/POLDIP2 reduces the sensitivity to lower concentrations of Ang II by >100-fold and tempol prevents all of these changes Thus, although a high salt intake reduces circulating Ang II, blockade of angiotensin receptors or ROS may prove beneficial for patients with CKD unable to restrict salt.
Abstract 057: BBSome in Smooth Muscle is Required for the Regulation of Vascular Function
Authors: Reho John J , Guo Deng-Fu , Morgan Donald , Rahmouni Kamal
Published: December 2018

The BBSome is a protein complex composed of 8 Bardet-Biedl syndrome (BBS) proteins, including
Bbs1
, involved in the regulation of various cellular processes including trafficking of receptors to cilia and the plasma membrane. This protein complex has emerged as a key regulator of cardiovascular function and blood pressure. We tested the hypothesis that the BBSome in vascular smooth muscle contributes to the regulation of vascular function. We selectively disrupted the BBSome in smooth muscle cells by deleting the
Bbs1
gene. We crossed mice harboring floxed alleles of the
Bbs1
gene (Bbs1
F/F
) with mice expressing tamoxifen-inducible Cre in smooth muscle cells (smMHC
CreERT2
). Validation of
Bbs1
gene deletion in smooth muscle was confirmed via decreased
Bbs1
mRNA levels in the aorta (1.0±0.2 vs 0.4±0.1 AU). We used wire myography to assess isolated aortic and mesenteric arterial ring function.
Bbs1
gene deletion from smooth muscle resulted in a significant decrease in the relaxation responses evoked by acetylcholine (Max relaxation: 55±7 vs 25±12%) and sodium nitroprusside (Max relaxation: 85±3 vs 61±8%) in aortic rings indicating dysfunction in both the endothelium and smooth muscle. Interestingly, aortic rings from SM
Cre
/Bbs1
F/F
mice exhibited enhanced contractility to endothelin-1 compared to controls (Max contraction: 0.1±0.01 vs 0.06±0.01g). Additionally, resistance mesenteric arterial rings of SM
Cre
/Bbs1
F/F
mice displayed enhanced contractile responses evoked by U-46619, phenylephrine, endothelin-1 and KCl-induced depolarization relative to controls (p<0.05). Western blot analysis revealed reduced expression of eNOS (1.0±0.1 vs 0.4±0.1 AU) and PKCα (1.0±0.2 vs 0.3±0.2 AU) in aortic lysates of SM
Cre
/Bbs1
F/F
mice compared to controls. Radiotelemetric mean arterial pressure was not different in SM
Cre
/Bbs1
F/F
mice at 4 weeks post-tamoxifen (96±4 mmHg) relative to controls (99±1 mmHg). Finally, arterial stiffness, measured using pulse wave velocity 4 weeks post-tamoxifen, showed a trend toward increased aortic stiffness in SM
Cre
/Bbs1
F/F
mice (2.6±0.1 m/s) compared to controls (2.3±0.1 m/s, p=0.06). These data point to the vascular smooth muscle BBSome as a novel regulator of vascular function in a manner independent of blood pressure.
Abstract 058: Novel Mechanisms at Immune-Vascular Interfaces Regulate Myogenic Tone of Resistance Arteries and Induce Hypertension in Mice
Authors: Carnevale Daniela , Iodice Daniele , Vinciguerra Iolanda , Perrotta Marialuisa , Lembo Giuseppe
Published: December 2018

Multiple reports recognized that immunity participates in hypertension (HTN), although direct mechanisms that orchestrate the immune-vascular interaction are still unknown. Mice infused with AngiotensinII (AngII) typically increase blood pressure (BP), concomitantly recruiting immune cells in the vasculature, with CD8 T cells playing a crucial role. To investigate the molecular regulators of vascular-immune interface, we established a system of vessel organ culture, enabling the study of physiological properties of resistance arteries via a modified pressure myograph which keeps vessels alive and functional for several days, while co-culturing immune cells of interest. We tested potential direct effects of CD8 T cells isolated from AngII-HTN or vehicle mice on resistance arteries from naïve mice. After co-incubation in the organ culture, CD8 T cells from HTN mice significantly increased myogenic tone (MT) of arteries, while no effect was induced by CD8 T of vehicle mice (%MT=CD8
AngII
:27±2; CD8
veh
:16±2, p<0.01). Our previous studies showed that an intracellular signaling involved in the acquisition of CD8 effectors functions, phosphatidylinositol-3-kinase-γ (PI3Kγ), was also involved in AngII-induced HTN, being its inhibition protective toward increase in BP and in vascular resistance. Here we took advantage of a mouse model expressing constitutively active PI3Kγ (PI3Kγ
CX
), to test immune functions of PI3Kγ in HTN. PI3Kγ
CX
showed a spontaneous hypertensive phenotype (SBP
mmHg
:129±1 vs WT:104±2, p<0.001) accompanied by infiltration of activated CD8 T cells in renal vasculature. Then, CD8 T cells isolated from PI3Kγ
CX
mice and placed in co-culture with WT arteries increased their MT. Conversely, the CD8 T cells from WT mice had no effect when cultured with vessels (%MT=CD8
CX
:31±1; CD8
WT
:19±1, p<0.001). To test the in vivo relevance, we performed an adoptive transfer of CD8
CX
in WT mice, finding that they developed spontaneous HTN, while no effect was induced by CD8
WT
(SBP
mmHg
: CD8
CX
128±2 vs CD8
WT
103±2, p<0.001). Taken together these data show that CD8 T cells from AngII-HTN mice are able to directly increase MT of resistance arteries and that PI3Kγ signaling is a crucial modulator of this effector function, likely contributing to BP increase.
Abstract 069: Antihypertensive Medication Treatment Regimens in SPRINT
Authors: King Jordan B , Moran Andrew , Derington Catherine , Herrick Jennifer , Kronish Ian , Stults Barry , Shimbo Daichi , Muntner Paul , Greene Tom , Cushman William , Weintraub William , Bress Adam
Published: December 2018

Background:
Details of antihypertensive medication regimens used to achieve intensive systolic blood pressure (SBP) goals have not been described.

Objective:
Determine the distribution and longitudinal changes in antihypertensive medication regimens in the Systolic Blood Pressure Intervention Trial (SPRINT).

Methods:
We used antihypertensive medication data collected by pill bottle review at each visit to categorize antihypertensive regimens by medication class. Free text string variables of medication names were independently reviewed by two clinical pharmacists to create standardized generic medication names and classes.

Results:
Figure 1 illustrates longitudinal changes in class combinations and number of drugs at the randomization, 6, 12, and 18-month visits. Fifty-six percent of participants modified their initial regimen by the 6-month visits; 43% of participants made additional modifications to their regimens from the 6-month to the 18-month visit. The most common initial regimens, and least likely regimens to be changed over time, were combinations with an ACEI/ARB and diuretics ± other classes (42% of initial regimens). Participants in the intensive arm added a mean (standard deviation) of 0.6 (0.9) medications to their initial regimens in the first 18-months compared to -0.1 (0.9) in the standard arm.

Conclusion:
Intensive blood pressure treatment requires more medication complexity in terms of class and dose. Further study of distinct regimens may reveal if certain class and dose combinations provide better SBP control, safety, or patient satisfaction.
Abstract 070: Adherence to Antihypertensive Drugs: Insights from the SPYRAL HTN Trials and Implications for Hypertension Trial Design
Authors: Weber Michael A , Kandzari David , Böhm Michael , Mahfoud Felix , Kario Kazuomi , Walton Tony , Hopper Ingrid , Patel Manesh , Tyson Crystal , Devireddy Chandan M , Lea Janice P , Weil Joachim , Agdirlioglu Tolga , Meyer Markus , Fahy Martin , Townsend Raymond
Published: December 2018

Background:
Variable adherence to prescribed antihypertensive drugs may have confounded prior renal denervation trials. Adherence to prescribed drugs was measured in the SPYRAL HTN-OFF MED and ON MED trials of renal denervation for uncontrolled hypertension when no drugs, and when up to 3 anti-hypertensive drug classes were prescribed, respectively.

Methods:
Patients were enrolled with uncontrolled hypertension defined as office systolic blood pressure (SBP) ≥150 and <180 mmHg with diastolic blood pressure >90 mmHg and 24-hour mean SBP ≥140 and <170 mmHg. Patients were randomized 1:1 to renal denervation or sham control. OFF MED trial patients were either drug naïve or had antihypertensive drugs discontinued at enrollment. ON MED trial patients had to maintain baseline prescribed antihypertensive medications. Both groups were informed on enrollment that urine and blood samples would be obtained for drug adherence testing. This testing utilized tandem HPLC and mass spectrometry. Following enrollment, both groups were followed for 4-8 weeks prior to randomization.

Results:
In OFF MED, antihypertensive drug or drug metabolites were detected in 10% of patients just prior to randomization, 11.2% at 3 months following randomization, and in 15% of patients at either time point, despite protocol-required absence of all antihypertensive medications following enrollment. In ON MED, 62.5% were fully adherent to prescribed medications (i.e., all prescribed medications identified) at baseline, 55% at 3 months and 62.5% at 6 months. Drug adherence was not consistent: 24% of baseline adherent patients were non-adherent at 3 months, and 16% of patients adherent at 3 months were no longer adherent at 6 months.

Conclusions:
Despite being aware that drug adherence testing was being conducted, almost half did not take all medication when they were prescribed and a minority took anti-hypertensive medications when instructed to abstain. Furthermore, non-adherence was inconsistent, with individual behavior at one time point not predictive of drug adherence at other time points. Attention to protocol adherence in device-based clinical hypertension trials is critical to determine non-drug treatment effects.
Abstract 071: Increased Short-Term Blood Pressure Variability in Patients With Isolated Systolic Hypertension
Authors: De La Sierra Alejandro , Parati Gianfranco , Banegas Jose R , de la Cruz Juan J , Gorostidi Manuel , Vinyoles Ernest , Segura Julian , Ruilope Luis M
Published: December 2018

Isolated systolic hypertension (ISH) is a clinical condition derived from increased arterial stiffness and associated with a worse cardiovascular prognosis. Increased blood pressure variability (BPV) is also associated to cardiovascular risk. We aimed to estimate short-term BPV in patients with ISH compared to those with systolic and diastolic hypertension (SDH). From the Spanish ABPM Registry database containing 63910 identified patients, we selected 20497 with ISH and 23269 with SDH. BPV estimates were obtained from ABPM records, and included standard deviation (SD), coefficient of variation (CV), weighted SD and average real variability (ARV). Comparisons were carried out by Student’s t test, and by general linear models adjusted for age, gender and office BP. Due to important differences in age between groups, a case-control matching was performed obtaining 14707 pairs matched by gender and with an age difference < 2 years. Comparisons were repeated in this dataset. Patients with ISH were older, more frequently women, with diabetes and with a previous history of CV disease. Office BP was lower (155±13/81±7 vs. 159±16/98±7 mmHg; p<0.001) in comparison to those with SDH. As shown in the table, systolic BPV was increased in ISH (p<0.001 for all comparisons), with respect to SDH. Results were confirmed after adjustments for age, gender and office BP and in the case-control matching comparison (also adjusted for office BP). No differences in diastolic BPV were observed between groups. We conclude that ISH is associated with an increased short-term BPV. This increased BPV could be one of the factors explaining the high CV risk of ISH patients.
Abstract 072: Return on Investment of Self-Measured Blood Pressure: An Economic Model from the Insurers’ Perspective
Authors: Arrieta Alejandro , Woods John , Wozniak Gregrory , Tsipas Stavros , Rakotz Michael , Jay Stephen J
Published: December 2018

Introduction:
About half of the 70 million adults with hypertension in the U.S. do not have their blood pressure (BP) controlled. An effective strategy to diagnose hypertension and improve BP control is to use self-measured blood pressure (SMBP) devices. Despite evidence supporting the
clinical
effectiveness of SMBP, most insurers remain unconvinced that the cost of the devices would yield a positive
financial
return.

Objective:
We adopted the perspective of private insurers to estimate return-on-investment (ROI) and net present value (NPV) of SMBP devices used to diagnose hypertension (including treatment selection and medication titration) and to manage BP.

Methods:
We developed a decision-analytic model using Framingham risk predictions and reported SMBP and clinic blood pressure measurement (CBPM) sensitivity and specificity values to simulate health outcomes and their associated annual and lifetime projections of costs and savings for the U.S. population.

Results:
Compared to CBPM, SMBP benefits exceed investment, producing large positive ROIs and NPVs. SMBP is cost-beneficial in the short-run and at lifetime horizon, but the return declines with patient age (see table).

Conclusions:
A strong business case exists for reimbursing SMBP, but only when it is used both to diagnose and to manage hypertension. Its primary economic value stems from its diagnostic role in ruling out white coat hypertension. If SMBP is used solely to manage (but not to diagnose) hypertension, our model indicates that its incremental effects on blood pressure reduction and cardiovascular event rates are not large enough to produce positive financial gains.
Abstract 075: Does the Benefit From Treating to Blood Pressure Targets Vary With Age?
Authors: Roush George , Singh Kevin , Zubair Ayla , Kostis William , Sica Domenic , Kostis John
Published: December 2018

Hypertension guidelines differ regarding how blood pressure targets vary with age. Crucial to this debate is how the benefit of treatment might vary with age. Therefore, a systematic search was conducted for trials randomizing treatment in the more intensive arm to systolic blood pressure (SBP) targets generally recommended, i.e., 140 mmHg or less. Meta-analysis examined the effects of age on risk reduction. Sixteen trials met criteria with SBP targets of 120-140 mmHg. Relative to high targets, low targets reduced risk for cardiovascular events (CVEs), 0.86 (95% CI 0.75,0.98), P=0.019, but treatment benefit differed among trials. This heterogeneity was explained by patient age. Relative to high targets, low targets reduced risk in older patients (mean ages 74+): 0.77 (0.61,0.97), P=0.025, but not in younger patients (mean ages 55-67): 0.90 (0.78,1.03), P=0.121, even though the latter had much greater statistical power. Risk reduction from low targets was greater in older patients than in younger patients: Relative risk in older patients versus relative risk in patients 16 years younger (2 standard deviations of age) = 0.78 (95% CI 0.63,0.98), P=0.036 (see Figure). Although generally not statistically significant, similar patterns were seen for specific CVEs and total mortality. Risks for adverse effects also tended to decrease with age. The number needed to treat to low targets to prevent 1 CVE in 10 years for 2 typical populations declined with age from 45 to 14 and from 110 to 13. In conclusion, for both relative risk and absolute risk, treating to SBP targets of 120-140 mmHg was more beneficial in older patients than in younger patients with no age-related increase in adverse effects.
Abstract 078: Chemerin Antisense Oligonucleotide Lowers Blood Pressure in a Rat Model of Adiposity-Associated Hypertension.
Authors: Ferland David , Darios Emma , Garver Hannah , Yeh Steve , Mullick Adam , Fink Gregory D , Watts Stephanie W
Published: December 2018

Chemerin is an adipokine, produced in liver and white adipose, whose human plasma levels have been positively associated with hypertension and obesity. In Sprague Dawley rats with normal adiposity and blood pressure (MAP), knockdown of chemerin throughout the body by antisense oligonucleotides (ASO) chronically reduced MAP by ~ 7 mmHg. Thus, we hypothesized that in high-fat-fed animals with hypertension and more visceral white adipose tissue, chemerin ASO would cause a greater reduction of blood pressure. Dahl salt-sensitive rats were fed a high fat diet (60% calories from fat) for 24 weeks after weaning. Radiotelemeters then were implanted for measuring MAP, temperature, heart rate and activity. After recovery and a one-week baseline recording period, animals received subcutaneous injections of either chemerin ASO (25 mg/kg) or scrambled control ASO (25 mg/kg) on days 0, 7, 14, 19. On day 21 the animals were euthanized for tissue and plasma analyses (chemerin PCR and Western). When compared to animals receiving scrambled control ASO, chemerin mRNA expression in animals receiving chemerin ASO was reduced by 99.5 ± 0.1% in liver, 90.6 ± 0.6% in mesenteric perivascular adipose tissue, and 99.3 ± 0.2% in retroperitoneal fat. Chemerin protein was undetectable in plasma from animals receiving chemerin ASO. MAP in rats receiving control ASO did not deviate significantly from baseline (179 ± 3 mmHg), while MAP in rats receiving chemerin ASO dropped by 32 ± 2 mmHg after four injections without a significant change in heart rate (abstract figure). These data support the involvement of chemerin in adiposity-associated hypertension and suggest a possible new approach to treating hypertension.
Abstract 083: Role of Alpha2a-Adrenoceptors in the Development of Hypertensive Nephropathy
Authors: Stegbauer Johannes , Rahman Masudur , Yakoub Mina , Hoch Henning , Vonend Oliver , Rump Lars C , Hering Lydia
Published: December 2018

Objective and Methods:
Hypertensive nephropathy is associated by increased renal sympathetic nerve activity. Alpha2A-adrenoceptors (alpha2A-AR) regulate sympathetic tone by controlling norepinephrine (NE) release by a negative feedback mechanism. Recently, we have shown that deletion of alpha2A-adrenoceptors increases renal sympathetic neurotransmission. To investigate whether deletion of alpha2A-AR play a role in the development of hypertensive nephropathy, we induced hypertension by infusing angiotensin (Ang)II (1000ng/kg/min) for 4 weeks in uninephrectomized wild-type (WT) and alpha2A-AR knockout (alpha2A-AR-/-) mice on an FVB background. Blood pressure was assessed by radiotelemetry.

Results:
NE release stimulated by renal nerve stimulation 5Hz was significantly higher in untreated kidneys of alpha2A-AR-/- compared controls. During AngII infusion (1000ng/kg/min) urinary NE excretion was significantly higher in alpha2A-AR-/- than control mice (4.3±0.6 vs. 2.1±0.4mg/dl creatinine,
P
<0.05). In line with an increased sympathetic NE release, the severity of hypertension caused by AngII infusion was significantly higher in alpha2A-AR-/- mice compared to control mice (176±4 vs. 163±3mmHg,
P
<0.01). Moreover, albuminuria (140±24 vs. 66±19mg/mg creatinine,
P
<0.05) as well as kidney injury assessed by glomerular injury score and renal fibrosis were accelerated and kidney function measured by cystatin C (1082±66 vs. 906±52ng/ml, P<0.05) was significantly decreased in alpha2A-AR-/- compared to WT mice. To test whether reduced renal sympathetic activity reversed this effect, renal nerve denervation (RDN) was performed in alph2A-AR-/- mice. RDN reduced renal NE content by 70%. In alpha2A-AR-/- mice, RDN reduced the severity of AngII induced hypertension 138±9 vs. 169± 11mmHg, P<0.05) and albuminuria compared to non-denervated alpha2A-AR-/- mice.

Conclusion:
Deletion of alpha2A-ARs exaggerates hypertensive nephropathy by increasing sympathetic NE activity. Reduction of NE release by RDN reduced blood pressure and albuminuria in alpha2A-AR-/- mice suggesting an important role of renal sympathetic tone in the pathogenesis of hypertensive nephropathy.
Abstract 091: Knock Out of
Pappa2
Enhances Blood Pressure Salt-Sensitivity and Renal Injury
Authors: Cowley Allen , Geurts Aron , Evans Louise , Kurth Theresa , Yang Chun , Zheleznova Nadezda , Wollner Clayton , Kumar Vikash
Published: December 2018

Pappa2 (pregnancy associated plasma protein A2) is a protease of insulin-like growth factor binding protein 5 (IGFBP5) which has been primarily studied in pregnancy and post-natal growth. Our recent studies using contiguous congenic strains indicate that
Pappa2
is linked to salt-sensitive hypertension in Dahl salt-sensitive (SS) rats. The role of
Pappa2
in cardiovascular and renal function has not been studied previously. We therefore mutated
Pappa2
in a salt-resistant subcongenic strain 26-P using the CRISPR-Cas9 method. Mutation of the
Pappa2
gene resulted in a 60-bp deletion and the absence of the wild type protein in the urine was verified by Western blot analysis. We
hypothesized
that deletion of
Pappa2
in the salt-insensitive strain would return or enhance BP salt-sensitivity as found in SS rats. Both parental rats and weaned offspring were maintained on a 0.4% NaCl diet until 9 weeks of age, then surgically instrumented with radiotelemeters for 24 hr BP recordings obtained while fed 0.4% NaCl and during 21 days of a 4% NaCl diet.

Results:
Mean 24 hr arterial pressure at 21 days of a 4.0% salt diet was significantly higher (P<0.05) in male
Pappa2-/-
(164
+
5 mmHg; n=11) compared to
Pappa2+/+
WT (137
+
4 mmHg; n=9) rats. Body weight (9 wks) of
Pappa2-/-
was less (P<0.05) than weight of
Pappa2+/+
WT rats (217
+
7 g vs 281
+
11 g). Although less than expected compared to previously studied SS rats, indices of renal injury were only moderately increased in
Pappa2-/-
compared to
Pappa+/+
WT rats including tubular protein casts (6.509 ± 0.789, n=11 vs 3.8 ± 0.718, n=9; P<0.05), cortical T-lymphocyte (CD3
+
/mm
2
; 96.1 ± 9.9, n=11 vs 70.9 ± 6.8, n=9; P=0.06), macrophage (CD68
+
/mm
2
; 74.6 ± 9.4, n=11 vs 47.6 ± 5.2, n=8; P<0.05), and interstitial smooth muscle actin (4.5 ± 0.4, n=11 vs 3.4 ± 0.2, n=8; P<0.05). In summary, the “rescue” of the BP salt-sensitivity trait with mutation of
Pappa2
provides the first direct evidence that this gene plays an important role in renal fluid and electrolyte homeostasis and the regulation of arterial pressure.
Abstract 096: Progesterone Upregulates Endothelial Mineralocorticoid Receptor Expression Which Predisposes Female Mice to Obesity-Induced Endothelial Dysfunction
Authors: Faulkner Jessica L , Kennard Simone , Antonova Galina , Bagi Zsolt , Jaffe Iris , Patel Vijay S , Belin de Chantemèle Eric J
Published: December 2018

Compelling evidence indicates a higher efficacy of mineralocorticoid receptor (MR) blockade for the treatment of cardiovascular disease in females with obesity and diabetes than in males, however, the origin of this sex-specific effect is unknown. We have shown that leptin induces endothelial dysfunction in obese female mice via aldosterone-dependent activation of MR and that only female mice develop endothelial dysfunction (vasorelaxation to acetylcholine) in response to aldosterone
ex vivo
. Therefore, we hypothesized that females express higher endothelial MR (ECMR) expression than males which predisposes females to obesity-associated endothelial dysfunction. RT-PCR analysis in isolated aortic endothelial cells of Balb/C mice revealed a higher NR3C2 (MR gene) expression in females compared to males (2.9±0.5-fold from male, P<0.05), however, no such difference was observed in non-endothelial cells. Similarly, human adipose tissue endothelial cells exhibited higher ECMR mRNA expression than males (2.1±0.1 fold from male). Female sex-hormone suppression (ovariectomy) decreased ECMR expression in female mice (-0.8±0.2-fold from sham), which was restored by progesterone supplementation (-0.1±0.1-fold from sham). NR3C2 mRNA gradually increased with progesterone in diestrous (1.6±0.1-fold from estrous) phase and increased further in pregnancy day 16 (9.2±0.2-fold from estrous). Furthermore, progesterone dose-dependently increased ECMR protein expression in human endothelial cells
in vitro
(P<0.05). Increases in ECMR associated with higher progesterone levels in diestrus and pregnant females were associated with an increased sensitivity to leptin-induced endothelial dysfunction in mice. In parallel, while leptin induced endothelial dysfunction in intact ECMR female mice (P<0.05), specific deletion of MR in endothelial cells protected female mice from leptin-induced endothelial dysfunction. These data indicate that progesterone drives the sex-difference in the levels of ECMR expression and predisposes female mice to leptin-induced endothelial dysfunction. In addition, these data provide a rational for the higher efficacy of MR blockade in obese and diabetic women suffering from cardiovascular disease.
Abstract 097: Interaction of miRNA, Endothelial Mineralocorticoid Receptor and Epithelial Sodium Channel in Endothelium Stiffness and Aortic Dysfunction
Authors: Jia Guanghong , Javad Habibi , Aroor Annayya R. , Yang Yan , Hill Michael A , Whaley-Connell Adam , Jaisser Frederic , Sowers James R
Published: December 2018

Excessive endothelial cell (EC) mineralocorticoid receptor (ECMR) and epithelial sodium channel activation in endothelium (EnNaC) increase oxidative stress and inflammation with associated cardiovascular abnormalities. Previous studies implicate elevations in circulating aldosterone (Aldo) in obesity enhance ECMR/EnNaC signaling that contribute to the development of vascular stiffness, in part, by reducing endothelial nitric oxide (NO) synthase (eNOS) activity and NO production. However, the specific role of ECMR/EnNaC signaling and its molecular mechanisms have not been explored. We hypothesized interactions between ECMR, dysregulation of miRNA expression, and EnNaC contribute to obese-/Aldo-induced endothelium dysfunction and aortic stiffness. Six week-old ECMR
-/-
and wild type littermate mice were fed a mouse chow or Western diet (WD) containing excess fat (46%) and fructose (17.5%) for 16 weeks. The EnNaC alpha subunit KO (EnNaC
-/-
) and EnNaC
+/+
female mice were administrated Aldo (250 μg/kg/day) via osmotic minipumps for 3 weeks beginning at 25-28 weeks of age. RNA sequencing showed that in ECMR
+/+
mice WD induced increased miR-7a, miR-34, and miR-6973a and reduced miR-99, miR-486a, miR-6904, miR-6916, miR-6240 that potentially target EnNaC expression. For EnNaC in vascular function, EnNaC
-/-
significantly reduced inward sodium currents by patch clamp in isolated mouse ECs. Meanwhile, EnNaC
-/-
significantly inhibited Aldo-induced endothelium stiffness and endothelium dependent relaxation observed in EnNaC
+/+
. Further, chronic Aldo infusion induced aortic endoplasmic reticulum stress, reduced eNOS activation, endothelium permeability, expression of pro-inflammatory cytokines IL1 and IL6, oxidative stress, and aortic collagen 1 deposition and these abnormalities were attenuated in EnNaC
-/-
mice. These data indicate that interaction of endothelial specific ECMR/miRNA/EnNaC mediates vascular endothelium dysfunction and prompts aortic stiffness.
Abstract 103: Acute, Dose-Dependent, Blood Pressure-Lowering Effect of Continuous Positive Airway Pressure in Autonomic Failure Patients With Supine Hypertension
Authors: Okamoto Luis E , Celedonio Jorge E , Diedrich Andre , Shibao Cyndya A , Gamboa Alfredo , Black Bonnie K , Paranjape Sachin Y , Robertson David , Biaggioni Italo
Published: December 2018

Primary autonomic failure (AF) is characterized by disabling orthostatic hypotension but half of these patients also have supine hypertension. We have previously shown that continuous positive airway pressure (CPAP 4, 8, 12 and 16 cm H
2
O, each for 2 min) applied during autonomic blockade with trimethaphan acutely decreased systolic blood pressure (SBP) in hypertensive and normotensive subjects (-17±2 and -8±2 mmHg at 16 cm H
2
O) due to decreases in cardiac output (CO) and stroke volume (SV). In this study, we hypothesized that CPAP would have an acute, dose-dependent, blood pressure-lowering effect in patients with autonomic failure with supine hypertension. Five CPAP levels (0, 4, 8, 12 and 16 cm H
2
O, each for 2-3 min) were applied sequentially to 15 AF patients with supine hypertension (12 males, 70±1 yr). Hemodynamic parameters were measured at the end of each CPAP level. We found that CPAP levels of 4, 8, 12 and 16 cm H
2
O significantly decreased SBP in a dose-dependent manner (Figure). The maximal SBP drop was -20±3 mmHg with CPAP 16 (from 154±4 to 134±4 mmHg, P<0.01), and was associated with a decrease in CO (-12±4%) and SV (-13+4%). Neither systemic vascular resistance nor heart rate changed significantly with CPAP. SBP returned to baseline levels within 5 minutes after removal of the CPAP (REC). We conclude that, in AF patients with supine hypertension, CPAP acutely decreases SBP in a dose-dependent manner, due to decreases in CO and SV likely reflecting a decrease in venous return. These results suggest that CPAP could be used as a non-pharmacologic approach to treat the supine hypertension of AF. Future studies are needed to test the safety and efficacy of this approach.
Abstract 105: PVN Inflammation Contributes to Brain GαI2 Protein Dependent Sympathetically Mediated Salt Sensitive Hypertension
Authors: Wainford Richard D , George Jonique
Published: December 2018

Hypothesis:
PVN inflammation contributes to salt sensitive hypertension (HTN) that develops following central Gαi2 protein down regulation.

Methods:
Male Sprague-Dawley (SD) rats receiving a continuous I.C.V. scrambled (S) or a targeted Gαi2 oligodeoxynucleotide (ODN) infusion (25μg/5μl/day) were fed a normal (NS; 0.6% NaCl) or high salt (HS; 4% NaCl) diet for 7-days (N=4-6/group). Additional groups received a S or Gαi2 ODN and minocycline (120 μg/day) co-infusion and a 7-day HS diet. On day 7 24h sodium balance, MAP, plasma norepinephrine content (via ELISA) and PVN specific mRNA levels of IL1β, IL6, IL10 and TNFα (via RT-PCR) were assessed. Separate groups underwent IHC analysis of PVN astrocytes and PVN microglia activation.

Results:
Central Gαi2 protein down regulation, but not control S ODN treatment, results in HS evoked HTN, sympathoexcitation, sodium retention, increased PVN mRNA levels of pro-inflammatory cytokines IL1β, IL6 and TNFα, decreased PVN anti-inflammatory 1L10 mRNA levels and no change in astrocyte PVN levels. Significantly, Gαi2 protein down regulation increased PVN microglial activation in rats on a NS diet. I.C.V. minocycline (anti-inflammatory antibiotic) co-infusion abolished the HS-evoked pro-inflammatory mRNA cytokine response and attenuated HTN, sympathoexcitation and sodium retention following central Gαi2 protein down regulation.

Conclusion:
PVN inflammation contribute to central Gαi2 protein dependent salt sensitive HTN by modulating sympathetic outflow and sodium homeostasis. We speculate central Gαi2 protein downregulation evokes microglial activation to prime a pro-inflammatory PVN response to HS intake.
Abstract 106: Central Angiotensin II Mediates Neurogenic Hypertension Through Post-Translational Regulation of Neuronal Nitric Oxide Synthase in the Paraventricular Nucleus of the Hypothalamus
Authors: Sharma Neeru M , Haibara Andrea S , Katsurada Kenichi , Liu Xuefei , Patel Kaushik P
Published: December 2018

Activation of renin-angiotensin- system, nitric oxide (NO) bioavailability and subsequent sympathoexcitation plays a pivotal role in the pathogenesis of many cardiovascular diseases including hypertension. Previously we have shown Ang II enhances PIN (a protein inhibitor of nNOS: neuronal nitric oxide synthase, known to dissociate nNOS dimers into monomers) mediated ubiquitination of nNOS in the PVN of rats with heart failure. To further elucidate the mechanism by which Ang II increases sympathetic outflow by regulating PIN mediated nNOS ubiquitination, we used Sprague-Dawley rats (250-300 g) subjected to intracerebroventricular infusion of Ang II (20 ng/min, 14days, 0.5μl/h) through osmotic mini-pumps and NG108-15 hybrid neuronal cell line treated with Ang II as an
in vitro
model. Ang II infusion increased baseline mean arterial pressure (126 ± 9* vs. 84 ± 4 mmHg) and renal sympathetic nerve activity (20.5 ± 2.3* vs. 6.4 ± 1.9 % of Max. Activity). Ang II infusion increased the expression of PIN (1.36 ± 0.04* Ang II vs. 0.81 ± 0.02 Veh) with a concomitant 52% decrease in dimeric nNOS and PIN-Ub conjugates (0.73 ± 0.04* Ang II vs. 1 ± 0.03 Veh) in the PVN. Further, Ang II-mediated increase in PIN expression(0.67±0.06* CHX AngII vs. 0.40±0.08 CHX 0h) was independent of CHX (protein synthesis inhibitor) mediated decrease in PIN expression (0.41±0.06* CHX AngII vs. 0.19±0.04 CHX 4h) suggesting Ang II-mediated post-translational stabilization of PIN. Substrate-dependent ligase assay in cells transfected with pCMV-(HA-Ub)8 vector revealed a reduction of HA-Ub-PIN conjugates after Ang II and proteasome inhibitor lactacystin (LC) treatment (4.5 ± 0.6* LC Ang II vs. 9.2 ± 2.2 LC). TUBE (Tandem Ubiquitin-Binding Entities) assay showed decrease PIN-Ub conjugates in Ang II-treated cells (0.82 ± 0.12* LC Ang II vs. 1.23 ± 0.05 LC) while AT1R blocker Losartan(Los) treatment diminishes the Ang II-mediated stabilization of PIN (1.21 ± 0.07 LC Los vs. 1.14 ± 0.04* LC AngII Los). Taken together, our studies suggest that increased central levels of Ang II contribute to the enhanced expression of PIN leading to reduced expression of the dimeric form of nNOS, thus diminishing the inhibitory action of NO on pre-autonomic neurons in the PVN resulting in an increase in sympathetic outflow.
Abstract 107: Neurons in the Organum Vasculosum of the Lamina Terminalis Sense NaCl and Angiotensin II to Regulate Sympathetic Nerve Activity and Arterial Blood Pressure
Authors: Stocker Sean D , Kinsman Brian J , Browning Kirsteen N , Wenner Megan M , Farquhar William B
Published: December 2018

The organum vasculosum of the lamina terminalis (OVLT) contains specialized neurons that sense changes in extracellular NaCl concentrations to regulate sympathetic nerve activity (SNA) and arterial blood pressure (ABP). However, OVLT neurons also abundantly express angiotensin II (AngII)-type I receptors. Therefore, we performed a series of in vitro and in vivo studies in adult male Sprague-Dawley rats to determine whether OVLT neurons sense both NaCl and AngII to impact SNA and ABP. First, in vitro whole-cell recordings revealed the majority of OVLT neurons (66%, 23/35) showed an increased discharge frequency to both +7.5mM NaCl (1.1±0.2 to 2.2±0.3Hz, P<0.001) and 100pM AngII (1.3±0.3 to 2.2±0.3Hz, P<0.001). Second, in vivo single-unit recordings demonstrate the majority of OVLT neurons (61%, 11/18) displayed an increase in cell discharge to intracarotid injection of 0.5M NaCl (3.6±0.6 to 9.0±1.6Hz, P<0.01) and 200ng AngII (3.2±0.7 to 9.2±1.9Hz, P<0.01). Third, optogenetic inhibition of OVLT neurons (rAAV2-CaMKII-eNpHR3.0-mCherry, 5x10
12
particles/mL, 561nm, 10mW) attenuated thirst responses to IV infusion of 2M NaCl (1.25mL/30min; Laser OFF: 6.1±0.5mL vs ON: 2.0±0.7mL, P<0.05) and AngII (Laser OFF 7.2±0.7mL vs ON: 2.2±1.2mL; n=5 per group, P<0.05). Fourth, OVLT injection (20nL) of 0.5M NaCl or 10pmol AngII increased SNA (121±6% or 117±5%, n=4/group; P<0.05 respectively) and mean ABP (7±1 or 9±2mmHg, n=4/group; P<0.05 respectively). To determine whether chronic activation of OVLT neurons produces hypertension, rats received an OVLT injection of rAAV2-hSyn-HM3D(Gq)-mCherry (3x10
12
particles/mL, 50nL) and given access to 0.1% NaCl chow and 0.9% NaCl drinking solution. After 4 weeks, daily administration of clozapine-N-oxide to the drinking solution (3mg/kg/day) significantly increased 24-h fluid intake (baseline: 60±10mL vs Day 7: 178±29mL; n=6, P<0.01) and 24-h mean ABP (baseline: 97±2mmHg vs Day 7: 108±1 mmHg; n=6, P<0.05). Ganglionic blockade with hexamethonium (30mg/kg, ip) produced a greater fall in mean ABP at Day 7 (baseline: -38±6 mmHg vs Day 7: -53±5 mmHg, P<0.05). These findings suggest OVLT neurons sense both NaCl and AngII to regulate body fluid homeostasis and SNA to produce a sympathetically-mediated hypertension.
Abstract 110: Vascular Smooth Muscle RhoBTB1 Protects From Hypertension and Arterial Stiffness by Cullin-3 Dependent Ubiquitination of Phosphodiesterase 5
Authors: Mukohda Masashi , Fang Shi , Wu Jing , Agbor Larry N , Nair Anand R , Ibeawuchi Stella-Rita C , Hu Chunyan , Liu Xuebo , Lu Ko-Ting , Guo Deng-Fu , Davis Debbie R , Keen Henry L , Quelle Frederick W , Sigmund Curt D
Published: December 2018

We previously reported that vascular smooth muscle cell (VSMC) selective expression of hypertension (HT)-causing mutations in either PPARγ or the E3 Ring Ubiquitin Ligase Cullin-3 causes nitric oxide resistance and HT. Here we sought to assess the physiological role of RhoBTB1, a VSMC PPARγ target gene and Cullin-3 substrate adaptor. S-P467L mice which selectively express dominant negative PPARγ-P467L in VSMC exhibit RhoBTB1-deficiency. We bred S-P467L mice with mice inducibly expressing RhoBTB1 in response to Cre-recombinase. Inducible VSMC-specific restoration of RhoBTB1 in S-P467L mice fully corrected the HT (SBP, 141±6 vs 124±3 mmHg, p<0.01, n=8-10), arterial stiffness (Aortic Pulse Wave Velocity, 3.8±0.2 vs 2.5±0.1 mm/ms, p<0.01, n=11-13), and vasodilator function (Aorta, 46±5 vs 80±2% ACh-induced relaxation, p<0.01, n=6-9). Notably, the cardiovascular protection occurred despite preservation of increased agonist-mediated contraction and RhoA/Rho kinase activity, suggesting RhoBTB1 selectively controls vasodilation. Sodium nitroprusside-induced production of cGMP in aorta was severely impaired in S-P467L but restored by RhoBTB1. Consistent with this, phosphodiesterase 5 (PDE5) activity in aorta was augmented 2.5±0.3 fold in S-P467L but was returned to normal by RhoBTB1. PDE5 and RhoBTB1 reciprocally co-immunoprecipitated in HEK293 cells. Ubiquitination of PDE5 by Cullin-3 in HEK293 cells was RhoBTB1-dependent. Consistent with a role for Cullin-3 in mediating turnover of PDE5, PDE5 activity was augmented in MLN4924-treated aorta, a Cullin inhibitor, and abrogated by PDE5 inhibitor. The beneficial cardiovascular effect of RhoBTB1 in S-P467L mice was phenocopied by PDE5 inhibition. Angiotensin-II infusion also causes RhoBTB1-deficiency and HT which was reversed by smooth muscle specific RhoBTB1 restoration. We conclude that RhoBTB1 augments the cGMP response to nitric oxide by restraining the activity of PDE5 by acting as a substrate adaptor delivering PDE5 to the Cullin-3 E3 Ring ubiquitin ligase complex for ubiquitination and proteasomal degradation. RhoBTB1 provides protection from HT, vascular smooth muscle dysfunction, and arterial stiffness in at least two models of HT.
Abstract 111: Endothelial Sodium Channel Activation Promotes Angiotensin II Induced Arterial Stiffness and Endothelial Dysfunction
Authors: Aroor Annayya , Jia Guanghong , Hill Michael , Habibi Javad , Yang Yan , Zhe Sun , Ramirez-Perez Francisco , Martinez-Lemus Luis , Whaley-Connell Adam , Jaisser Frederic , Sowers James
Published: December 2018

Arterial stiffness is an independent risk factor for the development and progression of cardiovascular and kidney disease. Inappropriate activation of the renin-angiotensin-aldosterone system (RAAS) is seen in obesity and contributes significantly to development of arterial stiffness, vascular dysfunction and hypertension. We have recently observed that aldosterone increases the presence of epithelial sodium channels on endothelial cells (EnNaC), which results in stiffening of the endothelial cortical surface (glycocalyx) of the aorta. Accordingly, we posited that deletion of the critical alpha catalytic subunit of EnNaC would decrease Ang II induced arterial stiffening and vascular relaxation responses, in part, via decreases in oxidative stress. To investigate, we used mice with EC cell specific deletion of EnNaC, by serially crossing floxed” EnNaC mice with Tie 2-Cre transgene mice. EnNaC expression was markedly decreased in ECs from EnNaC KO mice while macrophage expression of EnNaC was unaffected. Ang II (500 ng/kg/min) was administered by infusion to 9 month old wild (WT) and EnNaC KO mice for three weeks with or without administration of the antioxidant tempol (2mM/day) in drinking water. Whole cell Na+ currents in ECs, as evaluated by patch clamp, were significantly increased by Ang II infusion, and this was markedly attenuated in EnNAC KO and tempol treated mice. Further, aortic endothelial stiffness as determined by atomic force microscopy was significantly increased in Ang II infused mice and this increase was attenuated in EnNaC KO mice. EnNaC deletion also attenuated impaired acetylcholine (eNOS)-mediated vasorelaxation. Thus, treatment with the antioxidant tempol reduced Ang II induced vascular stiffness and prevented impairment of NO mediated vascular relaxation. Moreover, tempol administration to Ang II infused EnNaC KO mice had similar effects in both Ang II-tempol and Ang II-EnNaC KO groups. Arterial stiffness was significantly decreased by EnNaC deletion or treatment with the antioxidant tempol in femoral vessels as well. These results are consistent with an important role for oxidative stress in Ang II promotion of EnNaC activation, endothelial stiffness and impaired eNOS mediated vascular relaxation.
Abstract 126: Nox5 Regulation of Vascular Contraction Involves Oxidation of Endoplasmic Reticulum Calcium Channels and Calreticulin
Authors: Montezano Augusto C , Camargo Livia L , Carrick Emma , Rios Francisco J , Haddow Laura , Beatie Wendy , Holterman Chet E , Kennedy Christopher , Touyz Rhian M
Published: December 2018

The biological function of calcium-sensitive superoxide-generating Nox5 is unclear, but it may play a role in regulating contraction, as we previously demonstrated. Here we explored molecular mechanisms whereby Nox5 controls contraction. Human arteries and mice expressing human
NOX5
in smooth muscle cells (Nox5+SM22+) were studied. In arteries from hypertensive subjects, Nox5 expression, assessed by immunoblotting, was increased (50%, p<0.05 vs control). In human VSMCs, AngII-induced ROS generation (1 fold) and activation of myosin light chain (MLC) (2.5 fold) were exaggerated in VSMCs from hypertensive subjects (p<0.05 vs control); an effect that was attenuated by Nox5 siRNA. In arteries from Nox5+/SM22+ mice, contraction to U46619 was increased in (5.8±0.3 mN vs WT: 4.2±0.2 mN, p<0.05). These hypercontractile responses were inhibited by NAC (ROS scavenger), calmidazolium (calmodulin inhibitor), dantrolene (ryanodine receptor Ca
2+
channel inhibitors) and CDN1163 (SERCA channel activator), but not by a Nox1/Nox4 inhibitor (GKT137831). ONOO
-
levels were increased in vessels from Nox5+/SM22+ mice (5.8±0.9 vs WT 3.4±0.1 AU/mg, p<0.05). Inactivation of MYPT1 (181±1.8AU vs 164±1.9AU WT) and activation of MLC (207±10.3AU vs 155±2.7AU WT) were increased in VSMCs from Nox5+SM22+ (p<0.05). To assess the oxidative proteome in VSMCs, we immunoprecipitated reversibly oxidized proteins and observed oxidation of Nox5, decreased oxidation of MYPT1 and increased oxidation of SERCA2b in Nox5+/SM22+ mice . Proteome analysis of human VSMCs identified the ER Ca
2+
sensor, calreticulin, as a potential Nox5 binding protein. Calreticulin reversible oxidation was increased in VSMCs from Nox5+SM22+ mice and hypertensive subjects. Our study unravels crosstalk between oxidative stress and Ca
2+
in the vasculature, where Nox5 regulation of contraction involves ROS, Ca
2+
and endoplasmic reticulum localized Ca
2+
channels/proteins. We identify novel mechanisms whereby Nox5 influences pro-contractile signalling through processes involving oxidation of the ER Ca
2+
sensor, calreticulin, and ER Ca
2+
channels.
Abstract 128: Interleukin-17 Mediates Hypertension, Intrauterine Growth Restriction, Cytolytic Natural Killer Cells and Vascular Dysfunction in Pregnant Rats
Authors: Travis Olivia , White Dakota , Pierce William A , Ge Ying , Young Cassandra M , Williams Jan M , Spradley Frank , Cornelius Denise C
Published: December 2018

Preeclampsia is a hypertensive disorder of pregnancy characterized by intrauterine growth restriction (IUGR), vascular dysfunction, and chronic immune activation including increased T
H
17s and cytolytic NK cells (NK
C
). We recently developed a novel model of preeclampsia in which placental ischemia (PI)-induced T
H
17s cause a preeclampsia-like phenotype in pregnant rats characterized by hypertension, IUGR, oxidative stress (ROS), and increased cytolytic NK cells (NK
C
). In the current study we investigated a novel role for IL-17, the main cytokine secreted from T
H
17s, to directly induce IUGR, NK
C
activation, and vascular dysfunction in pregnancy. IL-17 (150 pg/day) was chronically infused into a subset of normal pregnant (NP) rats from gestation day (GD) 12-19 (NP+IL-17) via i.p. minipump. On GD 18 carotid catheters were implanted and on GD 19 MAP, fetal weight, placental weight, placental NK
C
, and NK
C
-associated proteins were measured and vascular reactivity of uterine arteries was assessed. Data are expressed as mean±SEM. MAP significantly increased from 100±3 mmHg in NP (n=9) to 115±1 mmHg in NP+IL-17 (n=12). Fetal weight significantly decreased from 2.5±0.04 g in NP to 2.3±0.03 g in NP+IL-17 (p<0.05). Placental weight significantly decreased from 0.62±0.02 g in NP to 0.55±0.01 g in NP+IL-17 (p<0.05). Placental ROS significantly increased 1353±337 RLU/min/mg in NP to 2210±180 RLU/min/mg in NP+IL-17 (p<0.05). Placental NK
C
increased from 2.6±1.6% of the total NK population in NP to 11.3±2.2% in NP+IL-17 (p<0.05). Placental granzyme B increased from 22.7±1.6 pg/mg in NP to 30.2±2 pg/mg in NP+IL-17 (p<0.05). Placental granzyme A increased from 3067±225 pg/mg in NP to 3926±210 pg/mg (p<0.05). Additionally, placental levels of VEGF, an important pro-angiogenic factor secreted by non-cytolytic uterine NK cells significantly decreased from 77.6±6.5 pg/mg in NP to 54.2±3.2 pg/mg in NP+IL-17 (p<0.05). We also observed impaired relaxation of uterine arteries in response to acetylcholine. These data suggest a shift from non-cytolytic NK to NK
C
cells in the placentas of NP+IL17. In addition to hypertension and ROS, this study demonstrates novel roles for IL-17 to directly mediate IUGR, NK
C
activation, and endothelial vascular dysfunction during pregnancy.