- Email: [email protected]
[312-POS]
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 5 (2015) 53–156
SNPs (i.e.: 744delA, and 395A/G) were investigated using allele-specific PCR. Expression of pluripotency markers (i.e.: Nanog, and Oct-4) and telomere length measurement were assessed using Real-Time PCR. Results: Real-Time PCR analyses demonstrated a significant down-regulation of Klotho ( 83%; p = 0.005) in patients with Preeclampsia versus Controls. Results of Western Blot agreed with Real-Time PCR ones. Polymorphism analysis results suggest that 744delA allele is associated with 3-fold increased risk for preeclampsia. Real-Time PCR investigation revealed a significant down-regulation of pluripotency markers in pathological group. Conclusions: Klotho expression is decreased in preeclamptic pregnancies. Further data are required to confirm the role of this protein in pathophysiology of preeclampsia and the possible link to long term outcomes. Disclosures: M. Cecati: None. S.R. Giannubilo: None. A. Corradetti: None. M. Emanuelli: None. F. Saccucci: None. A.L. Tranquilli: None. doi:10.1016/j.preghy.2014.10.316
[311-POS] Chronic early life stress programs the renin-angiotensin system components on white adipose tissue explants Vanessa Tabor a, Timothy Mahanes a, Analia Loria b (a University of Kentucky, Lexington, KY, USA, b Department of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, USA) Objectives: We previously reported that adult male rats exposed to maternal separation (MatSep), a model of early life stress, display enhanced responses to angiotensin II compared to controls (C, non-MatSep) later in life. Furthermore, 16-week high fat diet feeding induced greater fasting plasma glucose and insulin in MatSep rats. To tested the hypothesis that MatSep alters the expression of the reninangiotensin system components in white adipose tissue. Methods: White adipose tissue pads (perirenal: PR, epididymal: EP and subcutaneous: SQ, 200 mg) were dissected from adult MatSep and C rats fed a normal fat diet (n = 4). Tissue was incubated (37 °C, 1 h, 5% CO2 and air) with vehicle or AngII 10 5 M in DMEM media (2 ml, 2% BSA fatty acid free). Tissue was removed from the media and flash frozen. RNA was extracted and RT-PCR was performed to determine GAPDH, angiotensinogen (AGT), ACE1, ACE2, Neprilysin (NEP) and AT1, AT2 and MAS receptors (R). Results: mRNA expression of AGT and AT1R was greater in PR and SQ tissue from MatSep rats (6 fold and 3 fold, respectively, p < 0.05 vs. C). However, AT2R was downregulated in PR and SQ tissue from MatSep rats (no detectable vs. C). EP fat did not show differences in mRNA expression of AGT, AT1R and AT2R between groups. Although ACE1, ACE2 and MAS R displayed similar mRNA levels in PR, EP and SQ fat explants, NEP was greater in all-fat pads from MatSep rats (4 fold, p < 0.05 vs. C). AngII-induced increase in AGT and AT1R mRNA expression was similar in MatSep and C fat tissue (10 fold, p < 0.05 vs. vehicle).
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Conclusions: Increased AGT mRNA expression and AT1/ AT2R ratio may contribute to the metabolic disturbances observed in MatSep HFD-fed rats. Ongoing studies are focused on the analysis of the adipokines released in the media by the fat explants. Disclosures: V. Tabor: None. T. Mahanes: None. A. Loria: None. doi:10.1016/j.preghy.2014.10.317
[312-POS] Mechanisms of renal potassium retention during late pregnancy Crystal West, Emily Qiu, Chris Baylis, Michelle Gumz (University of Florida, Gainesville, FL, USA) Objectives: The fetus requires a large amount of potassium for normal development. To accommodate this need the normal pregnant rat accumulates considerable potassium over the course of gestation, most of which is retained during late pregnancy. This gestational potassium retention is essential for fetal development but the mechanism is unknown. The purpose of this study was to examine how renal potassium retention occurs in the setting of high circulating aldosterone and enhanced collecting duct sodium reabsorption in the pregnant rat. Recent work on the WNK kinases has provided interesting targets for divergent modulation of sodium and potassium handling in the distal nephron. Methods: We measured the mRNA expression of WNK1, WNK4, ROMK, H+/K+ – ATPase type 1 (HKA1), H+/K+ – ATPase type 2 (HKA2), and H+ – ATPase in the renal cortex, outer medulla, and inner medulla of virgin (V, n = 6), mid pregnant (MP, n = 6), and late pregnant (LP, n = 6) rats using quantitative real-time PCR. Results: We found an increase in HKA1 in the outer medulla in MP rats vs V and increased HKA2 expression in both cortex and outer medulla of LP rats vs. V. Furthermore, ROMK expression decreased in the inner medulla of MP and LP rats compared to V. The expression level of the other genes tested did not differ with pregnancy stage. Conclusions: The potassium retention of pregnancy may be due both to increased collecting duct potassium reabsorption (via increased HKA1/HKA2), as well as decreased expression of the potassium secretory channel ROMK, which would reduce secretion and excretion of potassium, despite increased sodium reabsorption. This regulation is regional with enhanced HKA2 in cortex/outer medulla and reduced ROMK in inner medulla. Future studies will determine the signaling pathways involved as well as protein levels and cellular localization of the tubular potassium transporters, to assess functional significance. Disclosures: C. West: None. E. Qiu: None. C. Baylis: None. M. Gumz: None. doi:10.1016/j.preghy.2014.10.318
SNPs (i.e.: 744delA, and 395A/G) were investigated using allele-specific PCR. Expression of pluripotency markers (i.e.: Nanog, and Oct-4) and telomere length measurement were assessed using Real-Time PCR. Results: Real-Time PCR analyses demonstrated a significant down-regulation of Klotho ( 83%; p = 0.005) in patients with Preeclampsia versus Controls. Results of Western Blot agreed with Real-Time PCR ones. Polymorphism analysis results suggest that 744delA allele is associated with 3-fold increased risk for preeclampsia. Real-Time PCR investigation revealed a significant down-regulation of pluripotency markers in pathological group. Conclusions: Klotho expression is decreased in preeclamptic pregnancies. Further data are required to confirm the role of this protein in pathophysiology of preeclampsia and the possible link to long term outcomes. Disclosures: M. Cecati: None. S.R. Giannubilo: None. A. Corradetti: None. M. Emanuelli: None. F. Saccucci: None. A.L. Tranquilli: None. doi:10.1016/j.preghy.2014.10.316
[311-POS] Chronic early life stress programs the renin-angiotensin system components on white adipose tissue explants Vanessa Tabor a, Timothy Mahanes a, Analia Loria b (a University of Kentucky, Lexington, KY, USA, b Department of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, USA) Objectives: We previously reported that adult male rats exposed to maternal separation (MatSep), a model of early life stress, display enhanced responses to angiotensin II compared to controls (C, non-MatSep) later in life. Furthermore, 16-week high fat diet feeding induced greater fasting plasma glucose and insulin in MatSep rats. To tested the hypothesis that MatSep alters the expression of the reninangiotensin system components in white adipose tissue. Methods: White adipose tissue pads (perirenal: PR, epididymal: EP and subcutaneous: SQ, 200 mg) were dissected from adult MatSep and C rats fed a normal fat diet (n = 4). Tissue was incubated (37 °C, 1 h, 5% CO2 and air) with vehicle or AngII 10 5 M in DMEM media (2 ml, 2% BSA fatty acid free). Tissue was removed from the media and flash frozen. RNA was extracted and RT-PCR was performed to determine GAPDH, angiotensinogen (AGT), ACE1, ACE2, Neprilysin (NEP) and AT1, AT2 and MAS receptors (R). Results: mRNA expression of AGT and AT1R was greater in PR and SQ tissue from MatSep rats (6 fold and 3 fold, respectively, p < 0.05 vs. C). However, AT2R was downregulated in PR and SQ tissue from MatSep rats (no detectable vs. C). EP fat did not show differences in mRNA expression of AGT, AT1R and AT2R between groups. Although ACE1, ACE2 and MAS R displayed similar mRNA levels in PR, EP and SQ fat explants, NEP was greater in all-fat pads from MatSep rats (4 fold, p < 0.05 vs. C). AngII-induced increase in AGT and AT1R mRNA expression was similar in MatSep and C fat tissue (10 fold, p < 0.05 vs. vehicle).
153
Conclusions: Increased AGT mRNA expression and AT1/ AT2R ratio may contribute to the metabolic disturbances observed in MatSep HFD-fed rats. Ongoing studies are focused on the analysis of the adipokines released in the media by the fat explants. Disclosures: V. Tabor: None. T. Mahanes: None. A. Loria: None. doi:10.1016/j.preghy.2014.10.317
[312-POS] Mechanisms of renal potassium retention during late pregnancy Crystal West, Emily Qiu, Chris Baylis, Michelle Gumz (University of Florida, Gainesville, FL, USA) Objectives: The fetus requires a large amount of potassium for normal development. To accommodate this need the normal pregnant rat accumulates considerable potassium over the course of gestation, most of which is retained during late pregnancy. This gestational potassium retention is essential for fetal development but the mechanism is unknown. The purpose of this study was to examine how renal potassium retention occurs in the setting of high circulating aldosterone and enhanced collecting duct sodium reabsorption in the pregnant rat. Recent work on the WNK kinases has provided interesting targets for divergent modulation of sodium and potassium handling in the distal nephron. Methods: We measured the mRNA expression of WNK1, WNK4, ROMK, H+/K+ – ATPase type 1 (HKA1), H+/K+ – ATPase type 2 (HKA2), and H+ – ATPase in the renal cortex, outer medulla, and inner medulla of virgin (V, n = 6), mid pregnant (MP, n = 6), and late pregnant (LP, n = 6) rats using quantitative real-time PCR. Results: We found an increase in HKA1 in the outer medulla in MP rats vs V and increased HKA2 expression in both cortex and outer medulla of LP rats vs. V. Furthermore, ROMK expression decreased in the inner medulla of MP and LP rats compared to V. The expression level of the other genes tested did not differ with pregnancy stage. Conclusions: The potassium retention of pregnancy may be due both to increased collecting duct potassium reabsorption (via increased HKA1/HKA2), as well as decreased expression of the potassium secretory channel ROMK, which would reduce secretion and excretion of potassium, despite increased sodium reabsorption. This regulation is regional with enhanced HKA2 in cortex/outer medulla and reduced ROMK in inner medulla. Future studies will determine the signaling pathways involved as well as protein levels and cellular localization of the tubular potassium transporters, to assess functional significance. Disclosures: C. West: None. E. Qiu: None. C. Baylis: None. M. Gumz: None. doi:10.1016/j.preghy.2014.10.318