bluuebunny/crossref_metadata_2025_split huggingface dataset:
sFlt1‐Induced Preeclampsia Enhances Cardiovascular Response to Post Partum Hypertensive Stimuli via Smooth Muscle Mineralocorticoid Receptor
Authors: Biwer Lauren , Lu Qing , Ibarrola Jaime , Stepanian Alec , Man Joshua , Carvajal Brigett , Camarda Nicholas , Zsengeller Zsuzsanna , Seely Ellen , Karumanchi S. A. , Jaffe Iris 
Published: May 2022

BackgroundPreeclampsia (PE), a syndrome of high blood pressure (BP) and renal damage in late pregnancy, is associated with increased soluble VEGF receptor (sFlt1) and survivors have increased risk of future hypertension and cardiovascular disease.HypothesisSince smooth muscle cell mineralocorticoid receptor (SMC‐MR) can be activated by hypertensive stimuli, we hypothesized that high sFlt1 exposure during pregnancy may induce a post‐partum state of enhanced vascular sensitivity via SMC‐MR activation.Methods/ResultsA PE model was induced by transient viral expression of sFlt1 in pregnant C57Bl6 mice. Elevated serum sFlt1, BP and glomerular endotheliosis was confirmed which all resolve post‐partum. In a small cohort of women with prior PE, we confirm salt sensitivity of BP. Thus, postpartum mice were implanted with telemetric BP monitors and exposed to repeated hypertensive stimuli (high salt and subsequent AngII). Mice with prior PE had a significantly increased BP response to hypertensive stimuli. Microvascular arteries from mice after PE and hypertensive stimuli had enhanced ex vivomyogenic tone and AngII vasoconstriction. To test the direct impact of sFlt1 on SMC‐MR function, cultured SMC (Pac1) were transiently exposed to sFlt1 (24 hours on then off) to mimic the post‐PE state. SMC‐MR transcriptional activity in response to aldosterone and AngII were significantly increased after sFlt1 exposure as measured by luciferase reporter assay. Target gene expression was also enhanced after sFlt1 exposure. Finally, the role of SMC‐MR in the post‐PE phenotype was tested in vivo by PE induction in SMC‐MR‐KO vs MR‐intact littermates. Post partum, SMC‐MR‐KO mice were protected from the PE‐induced increase in aortic stiffness, microvascular myogenic tone and AngII vasoconstriction.ConclusionsFlt1‐induced PE produces a state of enhanced sensitivity to stimuli that may be mediated by increased activation of SMC‐MR. These data support the use of MR antagonists to mitigate the increased risk of cardiovascular disease in women exposed to PE.
Mitochondrial‐derived ROS Contribute to the Development of Hypertensive‐Diabetic Kidney Injury
Authors: Wang Zhen , Fu Yiling , Sears Jaylan , da Silva Alexandre A. , do Carmo Jussara M. , Li Xuan , Mouton Alan , Omoto Ana Carolina M. , Hall John E. 
Published: May 2022

Hypertension (HTN) is common in diabetes mellitus (DM) and substantially increases the risk for diabetic nephropathy. Current therapies only slow the progression to end‐stage renal disease (ESRD) in hypertensive‐diabetic patients rather than halt it. Thus, new insights into the mechanisms of this disease are needed.To investigate the mechanisms by which HTN and DM interact to promote renal injury, we developed a model combining streptozotocin (STZ)‐induced type 1 DM and inter‐renal aorta constriction (AC)‐induced HTN in male Wistar rats. In this model (Wistar‐DM+AC), both kidneys are exposed to the same levels of hyperglycemia, circulating hormones, and neural influences, but the left kidney below the AC has normal to slightly reduced blood pressure (BP), while the right kidney above the AC is exposed to elevated BP. To assess the roles of mitochondrial‐derived reactive oxygen species (ROS) and mitochondrial dysfunction in causing kidney injury, Wistar‐DM+AC rats were also treated with saline vehicle or mito‐TEMPO, a mitochondrial‐targeted antioxidant, for 6 weeks starting 2 weeks after AC surgery (0.4mg/kg/day, osmotic mini‐pump). Eight weeks after AC or sham surgery, glomerular filtration rate (GFR) and urinary albumin excretion (UAE) in left and right kidneys were measured separately by collecting urine from each kidney to evaluate the change of renal function. Mitochondrial oxygen consumption rate and superoxide generation from the cortex of left and right kidneys were measured simultaneously by Oroboros high‐resolution respirometry.Kidneys exposed to DM or HTN alone had only mild glomerular injury and slightly increased UAE. In contrast, right kidneys from Wistar‐DM+AC rats which had been exposed to DM plus HTN for 8 weeks had much greater increases in UAE (3.4±0.7 vs1.9±1.3 μg/min), significantly reduced GFR (0.5±0.1 vs 0.7±0.1 ml/min/g of kidney weight) as well as reduced mitochondrial oxygen consumption rate and increased mitochondrial ROS generation compared to left kidneys of Wistar‐DM+AC rats. In Wistar‐DM+AC rats that received mitoTEMPO treatment, the right kidneys exposed to the same levels of elevated BP and blood glucose had lower albumin excretion (1.7±0.3 μg/min), higher GFR (1.3±0.2 ml/min/g of kidney weight), and preserved mitochondrial oxygen consumption with reduced mitochondrial‐derived ROS compared to the right kidneys of vehicle‐treated Wistar‐DM+AC rats.These results suggest that mitochondrial‐derived ROS may contribute to kidney injury when HTN is superimposed on DM. Pharmacological inhibition of mitochondrial ROS generation attenuates kidney injury and may be a potential therapeutic strategy for diabetic nephropathy.
Effects of Dietary Nitrate Supplements on Morning Blood Pressure Surge and Sympathetic Neural Control during Orthostasis in Post‐menopausal Women
Authors: Takeda Ryosuke , Yoo Jeung‐Ki , Curtis Bryon M. , Roberts‐Reeves Monique A. , Hissen Sarah L. , Fu Qi 
Published: May 2022

Post‐menopausal women are at greater risk for cardiovascular disease compared with age‐matched men. An exaggerated morning blood pressure surge (MBPS) in post‐menopausal women is well established as an independent risk factor for cardiovascular events. Dietary nitrate supplements such as nitrate‐rich beetroot juice (BRJ) may reduce blood pressure (BP), but no study has investigated the chronic effects of BRJ on MBPS and sympathetic neural control during orthostasis in post‐menopausal women. We hypothesized that dietary nitrate supplements would decrease MBPS and muscle sympathetic nerve activity (MSNA, microneurography) in post‐menopausal women.We performed a double‐blind, placebo‐controlled, cross‐over trial and randomly assigned healthy women [n=14, 64±3 (SD) yr] to either dietary nitrate supplements (140 ml of BRJ; 800 mg nitrate/day) or placebo (140 ml of nitrate‐depleted BRJ daily) for 2 weeks each with ≥2 weeks of washout. Twenty‐four‐hour ambulatory BP monitoring was performed and MBPS was calculated as the morning systolic BP (SBP; averaged for 2 h after wake‐up) minus the lowest nocturnal SBP before and after 2‐week BRJ or placebo intervention. BP, heart rate, and MSNA were measured during 3‐min supine rest and 5‐min 60° head‐up tilt (HUT). Cardiac output (Qc; C2H2 rebreathing) and total peripheral resistance (TPR=mean BP/Qc × 80) were also measured.MBPS decreased after BRJ intervention (Figure A; p=0.043). BRJ did not change hemodynamics or MSNA during supine rest (Figure B‐E; all, p>0.05). However, upright TPR was reduced (Figure C; p=0.007), while Qc trended greater (Figure B; p=0.072) after BRJ intervention. During HUT, MSNA burst frequency (53±15 pre vs. 48±10 bursts/min post, p=0.508 for posture × intervention interaction) and burst incidence (70±17 vs. 63±15 bursts/100 heart beats, p=0.199 for interaction) were similar, but mean burst area (Figure D; p=0.010) and total activity (Figure E; p=0.015) were lower after BRJ intervention. Placebo had no significant impact on hemodynamics and MSNA in the supine position or during HUT in post‐menopausal women (all, p<0.05).Collectively, dietary nitrate supplements decrease morning surge in BP and upright MSNA and vasoconstriction in post‐menopausal women. Whether reductions in sympathetic activation and peripheral vasoconstriction during orthostasis contribute to the reduced MBPS in these women remains to be determined.
Soluble Epoxide Hydrolase Inhibitor Prevents Homocysteine‐induced Cardiac Hypertrophy via Epoxyeicosatrienoic Acid–mediated Inhibition of TRPC3 Channels
Authors: Zhou Yang , Wang Xiang‐Chong , Sun Wen‐Tao , Xue Hong‐Mei , He Guo‐Wei , Yang Qin 
Published: May 2022

ObjectivesActivation of soluble epoxide hydrolase (sEH), a key enzyme catalyzing the degradation of epoxyeicosatrienoic acids (EETs), has been implicated in cardiac remodeling. We previously demonstrated the involvement of sEH and transient receptor potential isoform 3 (TRPC3) channels in homocysteine‐induced hypertrophy of cardiac cells. This study investigated the anti‐hypertrophic effect of sEH inhibitor with the aim to reveal the link between sEH/EETs and TRPC3 channels in cardiac hypertrophy associated with hyperhomocysteinemia.Materials and methodsMale Wistar rats on 10‐week standard or 2% (WT/WT) high methionine diet were given drinking water supplemented with or without the specific sEH inhibitor 1‐trifluoromethoxyphenyl‐3‐(1‐propionylpiperidin‐4‐yl) urea (TPPU, 5mg/L). In vitro, H9c2 cells were treated with homocysteine (100 μmol/L) in the presence or absence of TPPU (10 μmol/L) or 11,12‐EET (1 μmol/L). Cardiac remodeling was evaluated by echocardiography and histological analysis. Plasma homocysteine level and EETs content were determined by ELISA. Hypertrophic markers were detected by western blot and qPCR. TRPC3 channel current was recorded by patch clamp in whole‐cell mode.ResultsTPPU lowered the heart to body weight ratio, inhibited expressions of the hypertrophic markers atrial natriuretic factor (ANP) and β‐myosin heavy chain (β‐MHC), and reduced the size of cardiomyocytes in hyperhomocysteinemic rats. TPPU treatment ameliorated ejection fraction (85.50±0.87% vs. 74.18±2.19%, p<0.05) and fraction shortening (50.30±1.32% vs. 42.27±0.68%, p<0.01) of the hyperhomocysteinemic rats. Rats with hyperhomocysteinemia showed reduced levels of EETs and increased expression of TRPC3 channels, which were reversed by TPPU treatment. In vitro, TPPU and 11,12‐EET inhibited H9c2 cell enlargement induced by homocysteine, along with a downregulation of hypertrophic markers and TRPC3 channels. Homocysteine significantly increased TRPC3 channel current (4.74±0.49 vs. 1.76±0.21 pA/pF in control, p<0.001) and the enhancement was inhibited by TPPU (2.44±0.29 pA/pF, p<0.01).ConclusionsInactivation of sEH alleviates homocysteine‐induced cardiac hypertrophy and improves cardiac function. EET‐mediated inhibition of TRPC3 channels contributes to the protective effect of the sEH inhibitor TPPU.
LPAR2 receptor stimulates progression of gastric cancer through β‐catenin signaling pathway
Authors: Ara Hosne , Bhattarai Susmita , Sharma Sudha , Subedi Utsab , Yu Xiuping , Bhuiyan Md. Shenuarin , Miriyala Sumitra , Panchatcharam Manikandan 
Published: May 2022

IntroductionLysophosphatidic acid (LPA) is a simple lipid that has been implicated in cellular homeostasis and pathology of different cancer by binding with G‐protein coupled receptors (LPAR1‐6). The aberrant Wnt signaling pathway has been reported to facilitate a crucial role in tumorigenesis and therapy responses. Although dysregulated β‐catenin signaling pathway has been established as a critical mediator of gastric cancer, whether LPA and LPAR2 play a role in the β ‐catenin signaling pathway during gastric cancer progression remains elusive.ObjectiveOur present study explored an unknown role of LPA and LPAR2 in LPA‐induced activation of β‐catenin and its downstream signaling mechanism in β‐catenin mediated gastric cancer.MethodWe analyzed the mRNA levels of different LPA receptors in human gastric cancer by ULCAN database using TCGA dataset. LPA ELISA, Western blotting, and immunohistochemical analysis were done in the human normal stomach and cancerous stomach to measure the LPA level and check the expression of different LPA receptors. To investigate the functional role of LPAR2 in LPA‐mediated gastric cancer progression, we performed ECIS proliferation, migration assay, scratch assay, and transwell invasion assay in LPA or LPA with LPAR2 antagonist treated cells. To elucidate the molecular mechanism of LPA‐mediated gastric cancer progression, we performed Western blotting, Real‐time PCR, Luciferase assay, Immunocytochemistry, and nuclear fractionation in gastric cancer cells treated with LPA or LPA with LPAR2 antagonist.ResultWe confirmed a robust increase in LPA level and significantly increased expression of LPAR2 receptor in human gastric cancer tissue samples (P<0.001) by LPA ELISA, western blotting analysis, and immunohistochemistry, suggesting a possible role of LPA in gastric cancer through LPAR2 receptor. In cultured AGS and NCI‐N87 gastric cancer cell line, LPA treatment increased proliferation, migration, and invasion activity, whereas LPAR2 receptor antagonist abrogated the LPA induced effect. Further, when the cells were treated with LPA, it increased the β‐catenin activity, nuclear localization of β ‐catenin, and the expression of the downstream target genes of the β‐catenin signaling pathway. In addition, we confirmed LPA‐induced activation of the β‐catenin signaling pathway through the LPAR2 receptor by using LPAR2 antagonist and by knockdown of LPAR2 where LPAR2 antagonist treatment or LPAR2 knockdown rescued LPA induced activation of the β ‐catenin signaling pathway in both cell lines.Conclusionwe established a novel LPA‐LPAR2‐β‐catenin signaling pathway in the progression of gastric cancer. All our results support that LPAR2 is a potential marker and target for developing novel therapeutic strategies in the treatment of gastric cancer.
Investigating the influence of the gut microbiome on diabetic renal disease
Authors: Xu Jiaojiao , Sanchez Jason , Gupta Kunal , Pluznick Jennifer L. 
Published: May 2022

BackgroundGut microbiota have been implicated in a large array of pathophysiological processes including obesity and diabetes. In this study, we utilized a diabetic renal disease model with or without oral antibiotic treatment to investigate how gut microbiota alter diabetic renal disease.MethodsGut microbiota biomass was depleted using a modified antibiotic treatment protocol with a mixture of 1g/L ampicillin, 1g/L neomycin, and 0.5 g/L vancomycin (modified from Rakoff‐Nahoum et al. 2004). C57BL/6J male and female mice (6‐week‐old) were randomly assigned to four different groups (n=8 per sex per group): control diet (CD), high‐fat diet (HFD), CD + antibiotics, HFD + antibiotics. The mice were treated with different diets and/or with antibiotics in drinking water for nine weeks. We measured body weight (BW), non‐fasting glucose, insulin, and glomerular filtration rate (GFR). In addition, we performed a glucose tolerance test (GTT), insulin tolerance test (ITT), and blood analysis. Finally, we did H&E staining of the kidney, and analyzed urine Na+.Results1) BW was significantly increased by HFD (vs CD) in both males and females regardless of antibiotic treatment. 2) Non‐fasting glucose showed no difference among these four groups in males and females. 3) Fasting glucose was dramatically elevated by HFD in males but not females (vs CD). For the GTT, both the peak glucose level and area under curve (AUC) were significantly increased in HFD vs CD, regardless of antibiotic treatment or sex. 4) For the ITT, HFD promoted insulin resistance as measured by changes in the AUC in both males and females. For males on a CD, antibiotics improved insulin responsiveness. 5) GFR was increased by HFD vs CD in both males and females, consistent with a HFD‐induced hyperfiltration, as we published previously with this same model (Halperin Kuhns, AJP Renal 2018). Intriguingly, GFR in females was also enhanced by antibiotic treatment on both the CD and HFD (week 9: CD, F: 182±12.7; HFD, F: 290±19.9, p<0.0001; CD + antibiotics, F: 282±12.6; HFD + antibiotics, F: 373±20.1; p=0.0005 CD vs. CD+antibiotics; p=0.01 HF vs. HF+antibiotics). Similarly, GFR in males also increased with antibiotic treatment in CD, but, not HFD (week 9: CD, M: 191±10.0; HFD, M: 385±19.3, p<0.0001; CD + antibiotics, M: 263±10.5; HFD + antibiotics, M: 390±32.7; p=0.04 CD vs. CD+antibiotics; p>0.99 HF vs. HF+antibiotics). Preliminary data using germ‐free (GF) mice, which do not have microbiota, show that GFR was also increased in GF males and females (vs. conventional mice with gut microbes).ConclusionIn this study, depletion of the gut microbiota via antibiotics increased GFR in both males and females on CD. Interestingly, on a HFD, GFR increased with antibiotics in females but not males. These data suggest that gut microbial signals may directly influence renal function; we are now working to confirm these findings and to explore the underlying mechanisms.
Palisade Endings in Extraocular Muscle Exhibit Clear Motor Features but their Exact Function is still not known
Authors: Streicher Johannes , Carrero‐Rojas Genova , de la Cruz Rosa R. , Pastor Angel M. , Blumer Roland 
Published: May 2022

ObjectiveProprioception from extraocular muscles (EOMs) is important for proper visually‐guided behavior. Surprisingly, the classical proprioceptor pair (muscle spindles and Golgi tendon organs) is absent in the EOMs of most mammals and instead, palisade endings have been found in virtually all species analyzed so far. Due to their widespread distribution it is believed that palisade endings are candidates for EOM proprioception. We have analyzed the molecular profile and the central connection of palisade endings.MethodsWhole mount of cat EOMs were triple labeled with neuronal markers, including markers for cholinergic axons, exocytosis proteins, and acetylcholine receptors. In anterograde tracing experiments, neuronal tracer was injected into the EOM motor nuclei.ResultsPalisade endings were formed by nerve fibers that, coming from the muscle, extended into the tendon where they made a u‐shaped turn to approach the muscle tendon junction. Recurrent nerves divided into axonal branches and terminal varicosities to form palisade endings at the tip of single muscle fibers. Molecular analyses showed that palisade endings had a cholinergic phenotype because they expressed choline acetyl transferase, the synthesizing enzyme of acetylcholine. Additionally, palisade ending expressed SNAP25, synaptobrevin, synaptotagmin, complexin, and syntaxin, all of which are key proteins involved in neurotransmitter release. Acetylcholine esterase, the degrading enzyme of acetylcholine was absent in palisade ending and no receptors for cholinergic transmission were associated with palisade endings as demonstrated by the absence of alpha‐bungarotoxin signals. Following neuronal tracer injection in the EOM motor nuclei, palisade endings were labelled.ConclusionOur findings show that palisade endings are cholinergic, have an exocytosis machinery for neurotransmitter (acetylcholine) release and originate from the EOM motor nuclei. However, no receptors for cholinergic transmission are associated with palisade endings suggesting that palisade ending belong to an effector system that is different from that in other skeletal muscles.
Beta‐catenin inhibition as a novel therapeutic strategy for porphyria
Authors: Balogun Anu , Cornuet Pamela , Nejak‐Bowen Kari 
Published: May 2022

Background– The porphyrias are metabolic disorders caused by enzymatic defects in heme biosynthesis, leading to excessive accumulation of porphyrins and their precursors in the liver or bone marrow, the most active tissues in heme biosynthesis. The specific patterns of accumulation of the heme precursors aminolevulinic acid (ALA) and other intermediate compounds define clinical features of these diseases such as acute neurovisceral attacks, skin lesions, and mental changes. These debilitating diseases remain incurable, and there is an unmet need to develop effective therapies to treat them. Because the liver is either a source or sink for porphyrins, we investigated the role of Wnt/beta‐catenin pathway in this disease. Wnt/beta‐catenin signaling is an evolutionarily conserved pathway that plays an important role in liver health and pathophysiology by regulating genes important in metabolism and proliferation.Methods– 3,5‐Diethoxycarbonyl‐1,4‐dihydrocollidine (DDC) induces hepatic porphyria in mice and is utilized to identify signaling pathways involved in injury progression. The metabolism of DDC creates a potent inhibitor of ferrochelatase (FC), the terminal enzyme in the heme pathway, which causes the buildup of porphyrin precursors. These precursors can cause cellular abnormalities such as oxidative stress, mitochondrial dysfunction, protein aggregation and inhibition of crucial functions such as autophagy. We studied pharmacological inhibition of Wnt/beta‐catenin signaling by utilizing a beta‐catenin DsiRNA formulated into a lipid nanoparticle in mice fed DDC diet.Results– Our data demonstrates that mice with pharmacologically inhibited beta‐catenin have decreased liver injury due to significantly fewer porphyrin deposits. This is due to suppression of ALA‐S and ALA‐D enzymes which catalyze the first and second steps of heme biosynthesis respectively. Significantly, we identified a strong TCF4 binding site in the intron region of the Alad gene, implicating ALA‐D as a direct Wnt/beta‐catenin target. Our data, therefore, establishes a novel role of the Wnt/beta‐catenin pathway in regulating heme biosynthesis by inhibiting early steps in heme biosynthesis pathway and reducing toxic porphyrin accumulation. Furthermore, our data reveal that mice lacking beta‐catenin have increased induction of autophagy over baseline that contributes to the protection from injury by clearing accumulated toxic porphyrins.Conclusions– These observations collectively offer a novel opportunity to remedy porphyria by targeting the Wnt/beta‐catenin signaling pathway. Our next studies will further validate ALA‐D as the mechanistic target of beta‐catenin in the heme biosynthesis pathway. Because prior research has established crosstalk between autophagy and the Wnt signaling pathway, we will also investigate the role of beta‐catenin in regulating autophagy during porphyria.