bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Gemfibrozil on the Progression of Proteinuria in Obese Dahl Salt‐Sensitive Rats
Authors: Shields Corbin Anthony , Poudel Bibek , Browning Evan , Williams Jan Michael
Published: June 2021

The prevalence of obesity has increased dramatically within the last decade and is now considered an independent risk factor for chronic kidney disease (CKD). Recently, we reported that obese Dahl salt‐sensitive leptin receptor mutant (SSLepRmutant) rats exhibit dyslipidemia and renal injury independent of hyperglycemia that progresses to CKD. Interestingly, the kidneys from the SSLepRmutant strain displayed increased renal triglycerides and significant glomerular injury including podocyte foot process effacement and lipid droplets. Lipid‐lowering drugs such as fibrates have proven to be renoprotective in various models of kidney disease. Therefore, in the current study, we tested the effects of gemfibrozil, a fibrate, on the progression of renal injury in SSLepRmutant rats. Twelve‐week‐old SSWT and SSLepRmutant rats were separated into four groups (n=4 in each group): (1) SSWT and (2) SSLepRmutant rats treated with vehicle and (3) SSWT and (4) SSLepRmutant rats treated with gemfibrozil (200 mg/kg/day, orally, powdered food) for 4 weeks. Treatment with gemfibrozil had no effect on body weight or blood glucose levels in SSWT and SSLepRmutant rats. We observed greater than a 10‐fold increase in plasma triglyceride levels in the SSLepRmutant strain versus the values measured in SSWT rats (1265±380 and 117±19 mg/day, respectively). Gemfibrozil treatment significantly reduced plasma triglycerides by 64% in SSWT rats (41±20 mg/dL) and 57% in the SSLepRmutant strain (549±108 mg/dL). During the course of the study, proteinuria rose from 89±26 and 416±11 mg/day to 181±46 and 675±47 mg/day in SS and SSLepRmutant rats, respectively. Chronic treatment with gemfibrozil markedly decreased the progression of proteinuria by 52% in SSWT rats (87±17 mg/day) and 43% in the SSLepRmutant strain (385±75 mg/day). The kidneys from vehicle‐treated SSLepRmutant rats displayed significant glomerular injury with mesangial expansion and increased interstitial fibrosis and tubular protein casts compared to SSWT rats. Gemfibrozil treatment markedly decreased these renal abnormalities in both strains. These data indicate that reducing plasma lipid levels with gemfibrozil prevents the progression of proteinuria in both lean SSWT and obese SSLepRmutant rats.

Support or Funding Information

This study was supported by GM104357 and DK109133.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Increased Renal Expression of Adhesion Molecules and Inflammation in Diabetic Nephropathy
Authors: FAN LETAO , He Xiaochen , Muroya Yoshikazu , Fan Fan , Roman Richard J.
Published: June 2021

Diabetic nephropathy (DN) is the primary risk factor for end‐stage renal disease (ESRD). Previously, we reported that the T2DN rat develops hyperglycemia by 3 months of age, proteinuria by 6 months and progressive renal injury which closely mimics human diabetic nephropathy, including focal glomerular sclerosis, mesangial matrix expansion, thickening of basement membranes, and progressive renal interstitial fibrosis. However, the role of adhesion molecules and renal inflammation in the pathogenesis of DN in this model has not been studied. In the present study, we compared the expression of cytokine and adhesion molecules, as well as the infiltration of inflammatory cells in young (3‐month old), middle‐aged (12‐month old) and elderly (18‐month old) T2DN and SD rats. We found that the expression of the adhesion molecules, ICAM‐1 and E‐selectin, were elevated in elderly T2DN (1.3‐fold and 1.8‐fold, respectively), and E‐selectin level was already higher in young T2DN (2.1‐fold) than in age‐matched SD rats. The number of infiltrating CD45+ leucocytes, CD3+ T cells, CD3−/CD4+ monocytes and CD68+ macrophages per gram of kidney were increased by 2.81±0.45, 2.82±0.44, 3.06±1.05, and 7.86±3.75 times, respectively, in elderly T2DN versus SD control. The rise in infiltrating inflammatory cells was correlated with elevated proteinuria (1144±202 in T2DN vs. 293±47 mg/day in SD) and plasma creatinine level (1.01±0.1 mg/dl in elderly T2DN vs. 0.50±0.02 mg/dl in SD), a greater degree of glomerular sclerosis and a 2‐fold increase in renal fibrosis in elderly T2DN rats versus age‐matched SD rats. These results indicate that the increased expression of adhesion molecules promotes infiltration of inflammatory cells, which is associated with the progression of DN and suggests that immune modulatory drugs may slow the progression of this disease.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hypertension‐Induced Renal Injury is Associated with Impaired Glomerular Barrier Function Involving Podocyte Dysfunction
Authors: He Xiaochen , Chen Chun Cheng Andy , Murphy Sydney R. , Booz George W. , Roman Richard J. , Fan Fan
Published: June 2021

Dahl SS rats rapidly develop hypertension, proteinuria, and renal injury when challenged with a high‐salt diet. Previous studies revealed that a deficiency in the renal formation of 20‐HETE in SS rats impairs afferent arteriole myogenic response, autoregulation of renal blood flow, tubuloglomerular feedback and glomerular capillary pressure and enhances sodium transport in the thick ascending loop of Henle. However, the role of 20‐HETE in podocyte and glomerular barrier function is still unknown. The aim of this study is to investigate the role of 20‐HETE on the regulation of glomerular function and podocyte function using SS rats and CYP4A1 transgenic rats on a SS background. We found that the production of 20‐HETE was significantly higher in the isolated glomeruli of CYP4A1 compared to SS rats by using mass spectrometry. Immunoblot analysis demonstrated that the expression of α‐smooth muscle actin and vimentin was lower in the isolated glomeruli of CYP4A1 vs. SS rats, indicating reduced epithelial mesenchymal transition (EMT). After challenged with a 4% high‐salt diet, the mean arterial pressure increased to a similar extent in both strains as measured by radio telemetry. However, compared with SS rats, CYP4A1 rats developed lower proteinuria (127 ± 14 vs. 270 ± 23 mg/day, respectively), had unaltered glomerular permeability to albumin (Palb) by using a fluorescence dilution technique, and exhibited greater increases in Palb after treatment with HET0016, a 20‐HETE synthesis inhibitor. In addition, Masson's trichrome staining showed that the glomerular injury score was attenuated in hypertensive CYP4A1 transgenic vs. SS rats (1.3 ± 0.2 vs. 2.4 ± 0.2, respectively), and the area of renal interstitial fibrosis was significantly lower in CYP4A1 transgenic vs. SS rats (6% ± 1% vs. 18% ± 4%, respectively). Moreover, hypertensive CYP4A1 rats exhibited less podocyte foot process effacement and detachment than SS rats when examined by a transmission electron microscopy. These findings indicate that increased production of 20‐HETE has renoprotective effects in association with decreased EMT, preserved podocyte and glomerular barrier function. This study indicates that 20‐HETE is a potential novel pharmacological target and biomarker in hypertension and chronic kidney disease.

Support or Funding Information

This study was supported by grants HL36279 (RJR) and DK104184 (RJR), AG050049 (FF), P20GM104357 (RJR and FF) from the National Institutes of Health; 16GRNT31200036 (FF) from the American Heart Association.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Opioid Analgesics (Morphine, Fentanyl, and Sufentanil) and Trauma on Survival Time after Severe Conscious Hemorrhage in Rats
Authors: Klemcke Harold G , Calderon Mariam L , Rodriguez Cassandra M , Connor Brian S , Hinojosa‐Laborde Carmen
Published: June 2021

Background

Currently, opioid analgesics are not recommended for use on the battlefield for Soldiers with severe hemorrhage, although they are still used during such situations (Schauer et al., Prehosp Emerg Care 21: 744, 2017). Moreover, there have been few or no controlled, preclinical studies conducted using analgesic doses of fentanyl or sufentanil to support these guidelines. The current study was conducted to evaluate and compare effects of three opioid analgesics on survival after severe hemorrhage in conscious rats in the absence or presence of extremity trauma. We hypothesized that opioid drugs and trauma would both decrease survival time to severe hemorrhage.

Methods

Rats were randomly assigned to receive either 0.9% saline (S), 2.0 mg/kg morphine (M; 0.2 mg/100 μl), 10μg/kg fentanyl (F; 1 μg/100 μl) or 1μg/kg sufentanil (SuF; 0.1μg/100 μl). All rats (male; ~ 380grams) were surgically implanted with a carotid catheter. After 24 hours, rats were anesthetized briefly (10 min) to undergo soft tissue injury (crushing of the right gastrocnemius and semimembranosus muscles for 30 sec with forceps) and fibula fracture (T). Non‐trauma rats (NT) were anesthetized for 10 min only. Rats were allowed to awaken, and 90 min later underwent a conscious hemorrhage (~55% of blood volume) via the indwelling carotid catheter. At the end of hemorrhage, rats received either S, M, F, or SuF via the carotid catheter. Rats were observed for a maximum of 4 hr after the start of the 25 min hemorrhage.

Results

Within each trauma group, neither survival times (p > 0.51) nor percent survival (p > 0.55) differed among treatment groups (Table 1). However, when combined across all treatment groups, T rats had a shorter (p = 0.013) survival time (137 min, SD 81, n = 24) and lower percent survival (33.3%; p = 0.02) than did NT rats (196 min, SD 72, n=23, 69.6%).

Conclusions

At the analgesic doses given–previously shown to ameliorate behavioral indices of pain in rats (Xiang, J Trauma Acute care Surg 85: S49, 2018) –opioid drugs tested did not affect survival after severe hemorrhage. Exacerbating effects of extremity trauma on survival after hemorrhage in our rat model portend the need for rapid treatment of patients with traumatic injury accompanied by hemorrhage.

Support or Funding Information

Funding for this project was provided by the US Army CRMRP, JPC8, Applied Pain Research Programs, and USAMRMC.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Curcumin Does Not Attenuate the Preeclamptic Phenotype in the Dahl Salt‐Sensitive Rat
Authors: Ishimwe Jeanne A , Maeda Kenji J , Johnson Ashley C , Garrett Michael R , Sasser Jennifer M
Published: June 2021

Preeclampsia (PE) is associated with new onset of hypertension and proteinuria and affects 2–8% of pregnancies worldwide. It is a leading cause of maternal and fetal morbidity and mortality with no current therapy to manage the disease. While its etiology is yet to be elucidated, immune system activation of various inflammatory pathways is reported to be a contributing factor. Toll‐like receptor (TLR) activation by damage‐associated molecular patterns (DAMPS) released following poor placentation and oxidative stress has been linked to the development of PE and TLR4 expression is increased in the placenta of preeclamptic women. We performed a whole transcriptomic analysis on placental tissues from gestation day (GD) 14 rats using Affymetrix Rat 2.0ST GeneChip and noted significant dysregulation in multiple genes during pregnancy in the Dahl salt sensitive (S) rat, a model of spontaneous superimposed preeclampsia, compared to the normal pregnant control Sprague Dawley (SD) rat. These genes included upregulation of TLR4 in the placenta of Dahl S rats (1.9 fold, p<0.05), which was subsequently confirmed via quantitative real‐time PCR (2.9 fold, p<0.05). To further investigate this pathway, the goal of this study was to assess the effect of TLR4 inhibition in pregnant Dahl S rats. Curcumin, the anti‐oxidant and anti‐inflammatory active ingredient in the spice turmeric, has been reported to inhibit TLR4 activation, and reduced blood pressure in rodent models of LPS‐induced preeclampsia. We hypothesized that curcumin would inhibit TLR4 activation and reduce blood pressure and uterine artery resistance index during pregnancy in the Dahl S rat. Starting on gestational day (GD) 10, 17 weeks old rats were divided into two groups: a treated group (N=7) that orally received dough containing 400mg/kg of curcumin daily and a control group (N=6) that received the vehicle dough through GD20. On GD18, Uterine Artery Resistance Index (UARI) was measured via Doppler ultrasound. On GD20, terminal blood pressure measurements were taken, and pups and placentas harvested and measured. Curcumin had no effect on UARI (0.78 vs. 0.82, p=0.4) or mean arterial pressure (146 ± 5 vs. 133 ± 5 mmHg, p=0.08). Fetal outcome assessed by litter size (10 ± 1 vs. 11 ± 1 pups); pup weight (2.23 ± 0.08 vs. 2.30 ± 0.07 g) and pup length (29.44 ± 0.31 vs. 29.6 ± 0.46 cm) was similar between groups. These results suggest that administration of curcumin does not alleviate the preeclamptic phenotype in the Dahl S rat.

Support or Funding Information

R01HL1347111R01HL137673

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Connexin 37 contributes to water homeostasis and urinary concentrating ability
Authors: Xue Jianxiang , Rieg Jessica Ann Dominguez , Thomas Linto , Rieg Timo
Published: June 2021

Connexin 37 (Cx37) is one gap junction component which plays an important role in intercellular communication. In the kidney, multiple gap junction proteins have been localized including Cx37. Previous studies have shown that Cx37 is expressed in a punctate pattern in the renal vasculature and along the nephron. Regarding the latter, strong basolateral expression was found in the thick ascending limb and distal convoluted tubule, weaker expression was found in the proximal tubule and collecting duct. Functional data are limited, with one study implying a role of Cx37 in sodium homeostasis. Based on its localization, we hypothesized that Cx37 possibly contributes to fluid homeostasis. In order to gain further insight, we studied wild‐type (WT, n=9–12) and Cx37 knockout mice (Cx37−/−, n=9–12). Under baseline conditions, Cx37−/− showed a ~40% higher fluid intake (5.7±0.3 vs 4.2±0.3 mL/day, P<0.05) associated with a ~40% lower urine osmolality (1611±79 vs 2556±171 mOsm/kg, P<0.05) compared to WT mice. Plasma osmolality (Cx37−/−: 333±2 vs WT: 334±2 mOsm/kg, NS) and food intake (Cx37−/−: 4.0±0.2 vs WT: 3.7±0.2 g/day, NS) were not significantly different between genotypes indicating that Cx37−/− mice are in fluid balance. To determine the role of Cx37 in the urinary concentrating mechanism, we conducted 18‐hour water deprivation studies. In response to water deprivation, urine osmolality increased significantly in both groups but the increase remained ~375 mOsm/kg lower in Cx37−/− vs WT mice (3355±85 vs 3730±181 mOsm/kg, P<0.05), an effect associated with a more pronounced body weight loss (8.8±0.6 vs 7.2±0.5%, P<0.05) indicative of greater urinary water loss. The increase in plasma osmolality was not significantly different between Cx37−/− and WT mice (Δ change: 14±3 vs 11±2 mOsm/kg, NS). Consistent with greater thirst, fluid intake in the first 6 hours after water deprivation was significantly greater in Cx37−/− vs WT mice (4.1±0.4 vs 3.0±0.3 mL, P<0.05). In summary, these results demonstrate that expression of Cx37 is important for urinary concentration in vivo, including in response to water restriction.

Support or Funding Information

TR is supported by the National Institute of Diabetes and Digestive and Kidney Diseases (1R01DK110621). JX was supported by an American Heart Association Predoctoral Fellowship (18PRE33990236) and LT by a Postdoctoral Fellowship (19POST34400026).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Estrogen Modulates the Differential Expression of Cardiac Myocyte Chymase Isoforms and Diastolic Function
Authors: Wang Hao , Sun Xuming , Ahmad Sarfaraz , Su Jing , Ferrario Carlos M , Groban Leanne
Published: June 2021

Background and objective

Cardiac angiotensin II (Ang II) is involved in the diastolic dysfunction in postmenopausal women. Chymases, a family of serine proteases with chymotryptic activity, play a significant role in cardiac Ang II formation from its substrate Ang‐(1–12) in both human and rodent models. While human tissue expresses only the α‐form of chymase, α‐ and β‐forms exist in rodents. Five of the ten rat mast cell proteases (rMCP‐1–5) display chymase activity, and rats express predominantly the β‐chymase isoforms. This study was designed to assess the differences in enzymatic activity among these rMCP isoforms in Ang II formation, particularly in the cardiomyocyte (CM), and the effects of estrogen status on MCP gene expression and activities in female rat models.

Methods and results

Using conventional polymerase chain reaction (PCR) (Fig. 1A), real‐time PCR (Fig. 1B) and DNA sequencing (Fig. 1C), we demonstrated that MCP‐1, MCP‐2, MCP‐4, and MCP‐5 mRNAs are expressed in the CM of both spontaneously hypertensive rats (SHR) and normotensive Wistar‐Kyoto rats (WKY). While rMCP‐1 and rMCP‐5 gene transcripts were higher than other isoforms in both rat strains, WKY CM exhibits higher levels of rMCP‐1 and rMCP‐5 mRNAs compared to the SHR CM (Fig. 2). Ovariectomy (OVX) increased the expression of rMCP‐1 and rMCP‐5 mRNAs in WKY. In SHR, OVX was associated with a blunted increase in rMCP‐1 mRNA compared to OVX normotensive WKY. Chymase activity, measured by HPLC as Ang II formation from Ang‐(1–12), significantly correlated with rMCP‐1 and rMCP‐5 mRNA expression in both rat strains. Both rMCP‐1 and rMCP‐5 mRNA expressions were positively correlated with progressive diastolic dysfunction (increasing the ratio of early mitral inflow velocity‐to‐early mitral annular velocity, E/e′) and increases in left ventricular internal diameter end diastole.

Conclusion

These data show rMCP‐1 and rMCP‐5 as the Ang II forming chymase isoforms participating in diastolic dysfunction due to ovariectomy in rodents.

Support or Funding Information

This work was funded by grants from the National Heart Lung and Blood Institute (CMF and LG) P01‐HL051952 and the National Institute on Aging (LG) AG033727 of the National Institute of Health.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mast Cell Peptidases (Carboxypeptidase A and Chymase)‐Mediated Hydrolysis of Human Angiotensin‐(1‐12)
Authors: Ahmad Sarfaraz , Wright Kendra N , Varagic Jasmina , Groban Leanne , Ferrario Carlos M
Published: June 2021

Introduction

Angiotensin processing peptidases [carboxypeptidase A (CPA) and chymase] are stored in cardiac mast cell secretory granules and are co‐released into the extracellular environment after activation/degranulation. In the human heart, we showed that chymase is primarily responsible for angiotensin II (Ang II) generation from the alternate substrate angiotensin‐(1‐12) [Ang‐(1‐12)], which is present in human heart. In this study, we investigated whether there are any synergistic effects of CPA and chymase in the processing of Ang‐(1‐12).

Method

Radiolabeled human Ang‐(1‐12) [125I‐Ang‐(1‐12)] substrate was incubated with CPA alone (0.325 μg/mL), human recombinant chymase (hrChymase) (0.325 μg/mL) or a combination of CPA + hrChymase (1:1 or 1: ⅓ ratio) in 50 mM Tris‐HCl buffer solution containing 150 mM NaCl (pH 8.0) for 0–15 min at 37 °C. The reaction was stopped by adding an equal volume of 1% phosphoric acid and injected on a C18 column. Eluted 125I‐Ang‐(1‐12) metabolic products were quantified by HPLC connected to in‐line flow‐through gamma detector. To determine the Km and Vmax, CPA and hrChymase were incubated with increasing concentrations (0–300 μM) of human Ang‐(1‐12) substrate for 20 min at 37 °C. The Km and Vmax were calculated using the Michaelis‐Menten equation.

Results

We found that after 15 min of incubation, CPA sequentially metabolized Ang‐(1‐12) substrate into angiotensin‐(1‐9) [Ang‐(1‐9), 53% (major product)], Ang II (22%) and angiotensin‐(1‐7) [Ang‐(1‐7), 11%]. No further hydrolysis of Ang‐(1‐7) was detected during CPA incubation. In the presence of hrChymase alone, 125I‐Ang‐(1‐12) was directly metabolized into Ang II (89%) and no further hydrolysis of Ang II was detected. In the presence of both CPA + hrChymase (1:1 ratio), the amount of Ang II formation was 68% from 125I‐Ang‐(1‐12) within a 5 min incubation period. The formation of Ang‐(1‐9) and Ang‐(1‐7) were only 11% and 10%, respectively. When CPA + hrChymase were added in 1: ⅓ ratio for 5 min, the Ang II, Ang‐(1‐9) and Ang‐(1‐7) formations were 65%, 27%, 6%, respectively. The Km and Vmax values were 150 ± 5 μM and 384 ± 23 nM/min/mg of CPA and 40 ± 9 μM and 116 ± 20 nM/min/mg of hrChymase.

Conclusions

These findings suggest that compared to CPA, chymase shows a much higher affinity to metabolize Ang‐(1‐12) into Ang II. In addition, Ang II and Ang‐(1‐7) are the end products of chymase and CPA, respectively. Overall, our findings suggest that chymase, rather than CPA, is primarily responsible for Ang II generation from Ang‐(1‐12).

Support or Funding Information

This work was supported by grant from the National Heart, Lung and Blood Institute of the National Institutes of Health (P01 HL‐051952).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery of Bacteria in Meconium of Normal, Uncatheterized, and Uninfected Late‐Gestation Fetal Sheep
Authors: Doerr Beau E , Keller‐Wood Maureen , Wood Charles Evans
Published: June 2021

Fetal development is thought to proceed in a sterile environment. Recent reports of the presence of bacterial DNA in human placenta, the transfer of live bacteria from mother to fetus after hypoxia in the pregnant sheep, and the presence of bacteria in the meconium of newborn infants have suggested that the fetus might be exposed to bacteria in utero. We performed the present studies to build on our discovery of bacterial rRNA in the meconium of normal fetal sheep. Specifically, the present experiments were designed to test the hypothesis that intact bacteria could be visualized histologically in meconium from normal, uncatheterized, and uninfected fetal sheep. Pregnant ewes, uncatheterized and free of clinical disease, were euthanized in late gestation. At necropsy, meconium was isolated from fetal colon using RNAse and DNAse‐free instruments and snap‐frozen, then stored at −80C until analyzed. Approximately 150 mg of meconium was homogenized and both RNA and DNA extracted in Trizol. RNA was isolated, converted to cDNA using random hexamers, then subjected to endpoint PCR using primers 341F and 806R that amplify a 465 bp region of bacterial 16S rRNA. All assays were performed with both positive and negative controls. In meconium, 7/7 fetuses (143 days gestation; term=147 days) harbored bacterial RNA. Smears of meconium were stained using Gram stain or hematoxylin stain. Gram stain revealed Gram positive and Gram negative organisms with morphologies that are consistent with the genus/species of bacteria that we have previously identified in meconium using 16S rRNA gene sequencing in DNA isolated from meconium. Hematoxylin staining confirmed the morphology of bacteria observed in the Gram‐stained samples. Bacteria in meconium appear in clusters that are consistent with live, replicating, organisms. The present results support previous work from this laboratory demonstrating that bacteria gain access to the normal fetal sheep. These results are consistent with our observations that bacteria are present in low numbers in amniotic fluid. It is possible that amniotic fluid containing bacteria is swallowed by the fetus, that bacteria survive the transit through the immature fetal gastrointestinal tract, and that the bacterial population expands in the nutrient‐rich environment of the fetal colon.

Support or Funding Information

This work was supported by HD033053 and AI120195.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of a histone methyltransferase inhibitor on fertility in an animal model of endometriosis
Authors: Seguinot Inevy Ivtchelle , Craig Zellieann , Isidro Angel , Flores Idhaliz
Published: June 2021

Endometriosis is an inflammatory, painful gynecologic disease with no cure and limited treatment options. Epigenetics, including histone modifications, play a key role in the etiology of endometriosis, therefore it has been hypothesized that epigenetic drugs can be used as a non‐hormonal therapeutic alternative for endometriosis. Following up on our observation that treatment with a histone methyltransferase inhibitor (HMTi) significantly decreased disease parameters such as size of vesicles in an animal model of endometriosis, this study was designed to investigate the effects of HMTi on fecundity correlates (e.g,. changesin expression of fertility‐associated genes and in ovarian and endometrial histology). Two weeks after endometriosis induction in female Sprague Dawley rats, we injected rats intraperitoneally with HMTi or vehicle for four weeks. At sacrifice, the ovaries and uterus of the rats treated with HMTi or vehicle were collected. mRNA was extracted from the right ovary for qPCR of fertility associated genes (ER, LH, CYP19) and secondary and antral follicles were counted in the left paraffin‐embedded ovary. The uterus was paraffin‐embedded to evaluate histopathological parameters. Treatment did not modify the expression of any of the genes studied, and did not significantly affect the number of ovarian follicles compared to vehicle. In addition, HMTi did not cause any histopathological changes in uteri compared to vehicle. These observations suggest that HTMi does not negatively influence fecundity in this model, a possibility that is currently being investigated in vivo.

Support or Funding Information

Special thanks to our funding from Puerto Rico Science, Technology & Research Trust, PHSU RISE (NIGMS Grant R25GM062406), PHSU Magic Core & Brain Core (NIMHD Grant MD007579), Puerto Rico Biobank (NCI Grant 1U54CA163071), PRISE Program from University of Puerto Rico, Ponce Campus (NIGMS Grant R25GM096955) and Porter Physiology Development Fellowship.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Use of Photobiomodulation and Mesenchymal Stem Cells in Polycystic Ovary Syndrome‐Induced Rats
Authors: Alves Eduardo Donato , Benevenuto Luíz Guilherme Dércore , Amin Rebeca Gall , Donadoni João Antônio , Ferrari Kamile Bausells , Barros Michele Andrade , Achcar Jorge Alberto , Parizotto Nivaldo Antonio , Montrezor Luís Henrique
Published: June 2021

Evaluation was made of glycemia and gonadosomatic index (GSI) variations in rats with induced polycystic ovary syndrome (PCOS), following application of photobiomodulation (PBM) and injection of mesenchymal stem cells (MSCs). Eighty adult Wistar rats (n = 10 per group) were divided into control (C), PCOS, PBM, MSC, PCOS/PBM, PCOS/MSC, PBM/MSC, and PCOS/PBM/MSC groups. PCOS was induced by single injection of estradiol valerate (EV) (2 mg/kg/bw/i.m.). The animals were evaluated 30 and 60 days after induction. MSCs were injected directly into the ovaries (5×105cells/0.2 mL physiological solution in each ovary), on the same day as PCOS induction. The animals were irradiated with a laser (wavelength 808 nm, power 60 mW), using a dose of 3 Joules(J)/point on each ovary, 3 times weekly, totaling 6 J of energy for each irradiated animal per day of application. After sacrifice, plasma glucose was determined and the ovaries were removed and weighed for GSI determination (ovary weight/body weight × 100). Statistical analysis of the data was performed using ANOVA and Fisher's test. The results were reported as means ± SEM. This study was approved by the local Animal Care and Use Committee (CEUA–UNIARA, protocol nº 019/16). Glycemia: There were reductions in glycemia for all exposed groups, compared to the 30‐day C group (260.2 ±37 mg/dL), with the lowest values for the PCOS/PBM/MSC group (103.6 ±12.58 mg/dL). Compared to the 30‐day PCOS group (186.2 ±79.7 mg/dL), glycemia was lower for groups PBM/MSC (115.6 ±15.32 mg/dL) and PCOS/PBM/MSC (103. 6 ±12.58 mg/dL). After 60 days of treatment, the PCOS/MSC group presented lower glycemia (101.8 ±20.03 mg/dL), compared to the C group (160.6 ±42.3 mg/dL). 30‐days GSI: The PCOS/PBM(0.019 ±0.006%), PCOS/MSC (0.018 ±0.0061%), PBM/MSC (0.029 ±0.02%), and PCOS/PBM/MSC (0.027 ±0.008%) groups showed increased GSI, compared to the C group (0.012 ±0.0013%). 60‐daysGSI: There were increases of GSI for the PCOS/MSC (0.013 ±0.0048%), PBM/MSC (0.02 ±0.0035%), and PCOS/PBM/MSC (0.017 ±0.0036%) groups, compared to the C group (0.008 ±0.0039%). The results indicated that the PBM/MSC combination reduced glycemia 30 days after PCOS induction by EV. In addition, the combined use of PBMand MSC resulted in higher GSI, 30 days after PCOS induction, compared to the PCOS group, which was indicative of ovarian modulations induced by this treatment. Morphological, immunohistochemical, and hormonal studies are underway in our laboratory to complement the present study.

Support or Funding Information

Funding: FAPESP (Grant 2016/02811‐4)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High Intensity Focused Ultrasound Treatment Attenuates Disease Progression in a Mouse Model of Non‐Alcoholic Steatohepatitis
Authors: Huerta Tomas S. , Devarajan Alex , Tsaava Tea , Chavan Sangeeta S. , Tracey Kevin J.
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD), a widespread condition affecting approximately 35% of the US population, is characterized by excessive lipid accumulation in hepatocytes. Non‐alcoholic steatohepatitis (NASH) is a severe form of NAFLD that is characterized by hepatocyte inflammation alongside accumulation of fat in the liver. NASH can lead to cirrhosis and liver cancer. Recent studies suggest that activation of the inflammatory reflex that is mediated by vagus nerve can reduce the chronic inflammation often seen in metabolic disorders including NASH. Here, we utilize a preclinical model of NASH, which is induced by diet, to study the effectiveness of high‐intensity focused ultrasound (HIFU) in modulating specific hepatic neural networks and test HIFU as a therapeutic strategy. Mice in the NASH group were maintained on a high‐fat high‐carbohydrate (HFHC) diet consisting of 60% kcal from fat chow and sugar supplemented water (55% fructose, 45% sucrose), while the low‐fat diet (LFD) control group received 10% kcal from fat chow. After 8 weeks on these diets, animals received daily liver‐targeted HIFU or sham stimulation. Exposure to HIFU but not sham stimulation resulted in significant improvement in body weights (p > 0.05, t test) and blood glucose levels (p > 0.05 t test) in HFHC mice. Moreover, the body weight and blood glucose values of HIFU‐treated HFHC mice trended toward both LFD groups (HIFU‐treated and Sham‐treated). In addition, HIFU attenuated circulating pro‐inflammatory cytokines and metabolic disease markers in HFHC mice. HIFU improved glucose tolerance, reduced HOMA‐IR score, and resulted in less fat accumulation in the liver than Sham‐stimulated HFHC group. Collectively, these findings suggest that noninvasive HIFU stimulation of the liver is a potential therapy for the NASH phenotype. Further studies are necessary to elucidate the mechanism by which liver‐targeted HIFU stimulation reduces inflammation and hepatic fat accumulation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of a Probiotic and a Specifically Immunized Egg Powder on Blood C‐Reactive Protein in a Subclinical Population
Authors: Skelton Michele , DelCastillo Daniel , Music Gary , Owens Makayla , Tracey Megan
Published: June 2021

The purpose of this study was to examine the effects of a specifically immunized egg powder taken with a probiotic on blood C‐Reactive Protein (an inflammatory marker) in healthy subjects with subclinical GI complaints. In a randomized, double‐blind placebo‐controlled study, sixteen subjects were divided into two groups. One group received a probiotic plus 2 grams of a specifically immunized egg powder (treatment group) while the other group received a placebo. Blood C‐Reactive Protein (CRP) was measured prior to supplementation and immediately after a 4‐week supplementation phase. There was no change in the blood CRP for the placebo group. Blood CRP for the treatment group was reduced and approached significance (p=.07). The treatment group had a greater percent change in blood CRP compared to the placebo group (p < 0.05). Preliminary analysis suggests that a combination of a probiotic and a specifically immunized egg powder may contribute to lower blood CRP levels in healthy subjects with subclinical GI complaints.

Support or Funding Information

IgY Nutrition

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dietary TMAO Exerts Beneficial Effects in Heart Failure Rats
Authors: Ufnal Marcin , Huc Tomasz , Drapala Adrian , Samborowska Emilia , Konop Marek
Published: June 2021

Increased plasma trimethylamine oxide (TMAO) has been suggested to be a marker of cardiovascular risk. However, mechanisms of the increase and biological effects of TMAO are obscure. Plasma TMAO originates from gut bacteria‐produced TMA, and from dietary TMAO e.g. seafood.

Male, 6‐week‐old, Spontaneously‐Hypertensive‐Heart‐Failure rats (SHHF, SHHF/MccGmiCrl‐Leprcp/Crl, Charles River Laboratories, USA) and 4–5‐week‐old Spontaneously Hypertensive Rats (SHR, Medical University of Warsaw, Poland) were divided into Water‐groups and TMAO‐groups (drinking TMAO water solution 0.33g/L). Age‐matched Wistar‐Kyoto (WKY) were used as healthy controls. After one year of observation survival was 100% in WKY, SHHF‐TMAO, SHR‐Water and SHR‐TMAO but only 66% in SHHF‐Water. WKY showed no pathological changes in the circulatory system. SHHF showed heart failure with reduced systolic function and hypertrophic cardiomyopathy with histopathological picture of transition from hypertrophic to dilated cardiomyopathy. SHR showed heart failure with reduced diastolic function and hypertrophic cardiomyopathy. In comparison to SHHF‐Water, SHHF‐TMAO had 3‐fold higher plasma TMAO, a significantly lower sympathetic/vagal ratio, and a trend toward lower cardiac fibrosis. In comparison to SHR‐Water, SHR‐TMAO had 4–5‐fold increase in plasma TMAO, significantly lower plasma NT‐proBNP, significantly lower left ventricle end‐diastolic‐pressure and significantly lower cardiac fibrosis.

In conclusion, 3–5‐fold increase in plasma TMAO does not exert negative effects on the circulatory system in heart failure rats. In contrast, increased dietary TMAO seems to exert beneficial effect in heart failure, however, its magnitude may vary in different models or in different stages of heart failure.

Support or Funding Information

Supported by National Science Centre, Poland grant no. 2015/19/N/NZ5/00650 and 2016/21/B/NZ5/02544.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Western Diet‐Induced Metabolic Memory and Persistent Vascular Dysfunction in Female Rats
Authors: Persand Dhandevi , Sepulveda Maria Alicia Carrillo
Published: June 2021

Metabolic memory is a phenomenon by which the body remembers previous exposure to metabolic stressors, such as hyperglycemia. Consumption of a western diet (WD) is a major part of modern life contributing to the epidemic of Metabolic Syndrome (MS), a significant risk factor for cardiovascular disease. Recent studies from our laboratory have characterized a model of WD induced MS in female rats. If not treated early, MS often progresses to Type II Diabetes Mellitus (T2DM). Adopting a healthier diet prior to onset of T2DM may be sufficient to reverse the constellation of metabolic symptoms associated with MS including hyperglycemia, hypertriglyceridemia, and increased body fat around the waist. However, it remains unclear if this diet reversal overcomes metabolic memory and cardiovascular dysfunction associated with MS. We hypothesize that cardiovascular dysfunction will persist despite diet reversal due to the development of metabolic memory. Eight‐week old female Wistar rats were fed a WD (21% fat, 50% carbohydrate (34% sucrose), 20% protein) for 17 weeks (WD group) to induce MS. Then, the rats were subjected to diet reversal for 8 weeks (rWD group). The control group received a regular chow diet (5% fat, 48.7% carbohydrate (3.2% sucrose), 24.1% protein). Metabolic (body weight, triglycerides, free fatty acids, glucose tolerance) and cardiovascular (blood pressure and vascular reactivity) parameters were assessed. WD group showed increased body weight, triglyceride levels, and systolic blood pressure characterizing MS. Moreover, fatty liver was observed in the WD group. As expected, rWD group exhibited improvement of all metabolic parameters and reversal of fatty liver (Figure 1). WD group showed marked increases in systolic blood pressure, measured via right carotid catheterization. Using wire myography, cumulative concentration curves were performed for acetylcholine and sodium nitroprusside. We found a significant impairment of endothelium dependent and independent vasorelaxation, respectively. Surprisingly, the rWD group did not decrease blood pressure (Figure 2) or improve impaired vasorelaxation. These results indicate that WD causes long‐lasting cardiovascular complications that are not alleviated by diet reversal. Our findings show for the first time that exposure to a WD can lead to metabolic memory in the cardiovascular system. We will extend our studies to examine potential mechanisms of metabolic memory leading to persistent cardiovascular dysfunction.

Support or Funding Information

In‐house Grant NYIT

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Maternal exercise enhances brown adipose tissue development and thermogenesis in offspring mice challenged with high fat diet
Authors: Son Jun Seok , Liu Xiangdong , Chen Yanting , Tian Qiyu , Zhao Liang , Hu Yun , Chae Song Ah , Deavilla Jeanene , Zhu Mei‐Jun , Du Min
Published: June 2021

Accumulating evidences point to the early developmental origins of obesity and its associated metabolic diseases, which are prevalent. Brown adipose tissue (BAT) dissipates fatty acids and glucose as heat and, thus, enhancing BAT thermogenic function can effectively prevent obesity and metabolic symptoms. Physical exercise becomes less common in modern societies and moderate exercise during pregnancy is recommended, but its effects in BAT development and metabolic health of offspring have not been examined.

PURPOSE

To investigate the effects of maternal exercise on BAT development and thermogenesis, and metabolic health of offspring challenged with high fat diet (HFD).

METHODS

After mating, female C57BL/6J mice were randomly assigned into two groups of control (CON) and exercise (EX), and exercise was performed throughout gestation every morning for 60 min at moderate intensity. At birth, the litter sizes were normalized to 6. After weaning, offspring mice were fed HFD (60 kcal% from fat) during 8 weeks. The BAT were collected from both male and female offspring mice at weaning and also after 8 weeks of HFD feeding. Statistical difference was analyzed using student's t‐test and/or one‐way analysis of variance (ANOVA).

RESULTS

At weaning, there were increases in mRNA expression of BAT markers, Ucp1, Ppargc1a, Prdm16, Elovl3, and Cox7a1 and protein levels of UCP1, PGC‐1α, and PRDM16 in female and male offspring born to EX compared with offspring from CON (P < 0.05). In addition, surface temperature of EX offspring was higher than CON. After 8 weeks of HFD, EX mice gained less body weight, had higher insulin sensitivity, and lower hepatic lipid accumulation (P < 0.05) compared to CON mice. The oxygen consumption rate, mRNA expression of Ppargc1a, Elovl3, and Cox7a1, and protein levels of UCP1, PGC‐1α, and PRDM16 remained higher in female and male EX offspring (P < 0.05).

CONCLUSIONS

Exercise during pregnancy enhanced BAT development and thermogenesis in offspring, which protected offspring from obesity and metabolic syndromes when they consumed HFD, common in Western societies. Our data suggest that the lack of exercise of healthy women during pregnancy can predispose offspring to obesity and metabolic syndromes partially through impairing BAT function.

Support or Funding Information

Supported by NIH Grant R01HD067449.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mice Exposed to Early Life Stress Display Sex‐Specific Upregulation of Leptin Gene Expression in Adipose Tissue
Authors: Leachman Jacqueline Renee , Rea Matthew , Ritter Celia , Xu Xiu , Dalmasso Carolina , Fondufe‐Mittendorf Yvonne , Loria Analia S
Published: June 2021

Epidemiological studies have established early life stress (ELS) as an independent risk factor for increased BMI and cardio‐metabolic disease later in life. We have previously shown that a mouse model of ELS, Maternal Separation and Early Weaning (MSEW), increases adiposity in weanlings and exacerbates high fat diet (HFD)‐induced fat expansion in a sex‐specific manner. Therefore, the aim of this study was to investigate the sex‐specific effects of MSEW on adipocyte‐derived production of important hormones for body weight homeostasis, such as leptin and adiponectin. MSEW was performed by separating the pups from the dam for 4 to 8 hours during postnatal days (PD) 2 to 16, and weaned on PD 17. Control mice remained undisturbed and were weaned on PD 21. Following, eight‐week‐old mice were then fed on either a low fat diet (LFD, n=6) or HFD (10 or 60 % Kcal from fat, n=6) for 12 weeks. Each litter was represented by one mice of each sex randomly assigned to each diet. Body composition trajectory was assessed by EchoMRI. At the end of the experiment, mice were sacrificed for adipose tissue collection. Leptin and adiponectin gene expression was performed in gonadal white adipose tissue (gWAT) and subcutaneous WAT (scWAT). After 12 weeks of HFD, male MSEW mice showed increased BW compared with controls (47.9 ± 1.1 vs. 40.2 ± 2.6 g, p<0.05) as well body adiposity (36.7 + 1.1 vs. 22.2 ± 4.1 % body fat, respectively). In female MSEW mice fed a HFD, but not in males, the gene expression of leptin in gWAT was significantly upregulated compared with controls (5.0 ± 0.5 vs. 3.6 ± 0.5, p<0.05), but not in scWAT (2.1 ± 0.6 vs. 1.5 ± 0.5, p<0.05). Despite sex‐specific changes in adiponectin gene expression in response to HFD, there was not a significant effect due to MSEW. To begin to address the mechanism by which MSEW increases leptin expression in a sex‐specific manner, we analyzed the site‐specific methylation of the leptin promoter in gWAT from mice fed a HFD (n=4). The methylation landscape in the leptin promoter was different between male and female mice. However, we found that CpG sites 3 and 15 were significantly hypomethylated only in fat from female MSEW mice comparted to controls (site 3: 21±1 vs. 52±1 %5mC, p<0.05; site 15: 84±1 vs. 100 %5mC, p<0.05). Both sites in the leptin promoter are strong areas for transcription factor binding. In addition, DNMT3A was upregulated in MSEW mice (p<0.05). Taken together, our current working hypothesis is focused on elucidating whether exposure to MSEW may promote the binding of specific transcription factors that upregulate leptin gene expression in a depot and sex‐specific manner.

Support or Funding Information

R01HF135158 to ASL, R01HL135158‐01A1S1 to JRL

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The role of T cells on the elevated blood pressure of female and male PCOS offspring
Authors: Maranon Rodrigo Oscar , Barnes Allison , Machado Lorena , LaMarca Babbette , Reckelhoff Jane F.
Published: June 2021

Prenatal factors may program the immune response in offspring of women with polycystic ovary syndrome (PCOS) and contribute to their elevated cardiovascular and renal risk. For example, PCOS women might have altered T cell concentrations which could adversely affect their offspring; however, whether adult PCOS offspring have an increase in T cells remains undetermined. We hypothesize that T cells contribute to the increase in blood pressure and proteinuria in both male and female offspring from hyperandrogenemic female (HAF) rats. Female (f) and male (m) HAF offspring (HAF‐off) and placebo offspring (Pla‐off), 24 weeks old, from HAF dams (DHT pellet 7.5mg/90d starting at 4 wks of age) or placebo dams (placebo pellet/90d) were divided into 4 groups (n=4/grp): fPla‐off, fHAF‐off, mPla‐off, mHAF‐off, respectively. We measured fat mass and lean mass by ECHO MRI, body weight, proteinuria (24h urine), and mean arterial pressure (MAP) by telemetry. We also examined the circulating and intrarenal T regulatory (CD4+CD25+FOXP3+) or Th17 effector cells (CD4+CD25‐RORγ+) by flow cytometry. MAP and proteinuria were significantly higher in male and female HAF‐off than Pla‐off (mPCOS: 121±2 mmHg vs. mPla: 112±1 mmHg; p<0.05; fPCOS: 117±3 vs. fPla‐off: 109±2; p<0.05) (proteinuria: fPla‐off: 4.1±1 mg/24h vs. fHAF‐off: 9.4±1.7 mg/24h; p<0.05; mPla‐off: 20±2.2 mg/24h vs. mHAF: 43.7±6 mg/24h; p<0.05). Female and male pups of HAF dams have similar body weights as pups of control dams. However, males of HAF dams have greater body fat mass and less lean mass, than do males of control dams (mHAF‐off: 29.57±1.07 g vs. mPla‐off: 25.95±1.33 g; p<0.05, and mHAF‐off: 381.4±4.9 g vs. mPla‐off: 366.3±4.1 g; p<0.05, respectively). Females of HAF dams have similar fat mass but less lean mass than do control female offspring (LM: fPla‐off: 223.3±2.5 g vs. fHAF‐off: 214.9±2.0 g; p<0.05). Contrary to our hypothesis we did not find differences in circulating or intrarenal T cells between fPla‐off and fHAF‐off or mPla‐off and mHAF‐off. The exposure to hyperandrogenemia in utero life may not increases the T cells in PCOS offspring later in life and is not responsible for the increased BP. However, further preclinical studies are necessary to determine the role of the immune response and the possible mechanisms responsible for the increase in blood pressure and renal injury in PCOS offspring.

Support or Funding Information

Supported by AHA14POST18640015 (ROM) and P20GM121334 (ROM and JFR).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.