bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reactive oxygen species modulates the molecular targets of NBS‐1120 in pancreatic cancer
Authors: Santiago Gabriela , Chattopadhyay Mitali , Kodela Ravinder , Nath Niharika , Kashfi Khosrow 
Published: June 2021

BackgroundPancreatic cancer has poor survival rates and largely ineffective therapies. Aspirin is the prototypical anti‐cancer agent but its long‐term use is associated with significant side effects. NBS‐1120 belongs to a new class of “enhanced NSAIDs” that were designed to be safer alternatives by releasing nitric oxide and hydrogen sulfide. NBS‐1120 is built on an aspirin scaffold and while it retains all the classical pharmacological/therapeutic activities of aspirin, it is devoid of GI side effects. In this study, we evaluated the effects of NBS‐1120 against pancreatic cancer using cell lines and a xenograft mouse model.MethodsCell lines: MIA PaCa2 and BxPC3 human pancreatic cancer, ACBRI 515, human normal pancreatic epithelial. Cell kinetics: growth by MTT; Cell cycle phase distribution and apoptosis: Flow cytometry; Proliferation: PCNA. mRNA: qRT‐PCR. ROS: measured hydrogen peroxide and super oxide by flow cytometry, using DCFDA and DHE dyes. Xenografts: Male athymic nude mice (N=10) implanted s.c. in the right flank with MIA PaCa2 cells, after 10 days the mice were randomly divided into 2 groups (N=5/gp) and gavaged daily with NBS‐1120 (100 mg/kg) or vehicle. Tumor volume and animal wt were recorded every 3 days. After 30 days, mice were sacrificed, tumors excised, weighed, and fixed in 10% buffered formalin for IHC studies.ResultsNBS‐1120 inhibited growth of MIA PaCa‐2 and BxPC‐3 pancreatic cancer cells with IC50s of 52 ± 4, and 62 ± 5 nM, respectively, while it did not inhibit the growth of a normal pancreatic epithelial cell line at these concentrations. NBS‐1120 effects on cancer cell growth appear to be cyclo‐oxygenase (COX) independent as MIA PaCa2 cells are COX null while BxPC3 cells express both COX‐1 and COX‐2. NBS‐1120 inhibited cell proliferation, caused G0/G1 phase cycle arrest, leading to increased apoptosis. Treated cells displayed increases in reactive oxygen species (ROS) and caspase‐3 activity. In MIA PaCa‐2 cell xenografts, NBS‐1120 significantly reduced tumor growth and tumor mass, growth inhibition was due to reduced proliferation (decreased PCNA expression) and induction of apoptosis (increased TUNEL positive cells). ROS, iNOS, and p53 were increased, while NF‐κB and FoxM1 that were activated in untreated xenografts were significantly inhibited by NBS‐1120. Reduced FoxM1 protein expression was associated with reduced FoxM1 mRNA levels and increased p21 mRNA as measured by qRT‐PCR, likely explaining cell proliferative inhibition.ConclusionsNBS‐1120 preferentially affects cancer cells, targeting parameters important in determining cellular mass, and inflammation, and merits further evaluation.Support or Funding InformationSupported in part by the National Institutes of Health [R24DA018055; R01GM123508].This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Selective Sphingosine‐1‐Phosphate Receptor‐1 Ligand Protects Against Ischemia‐Induced Inflammation and Dysfunction in Human Brain Microvascular Endothelial Cells
Authors: Li Yujing , Shi Samuel , Liu Qiang , Shi Fu Dong , Gonzales Rayna J 
Published: June 2021

Cerebrovascular inflammation and endothelial cell dysfunction are key contributors to brain damage following acute ischemic stroke. In preclinical and clinical studies, sphingosine‐1‐phosphate receptor (S1PR) ligands are emerging as exciting therapeutic strategies to treat acute ischemic stroke. We previously observed that selective S1PR1 modulation reduced lesion volume following transient middle cerebral artery occlusion (tMCAO). However, S1PR1 expression and the influence of a selective S1PR1 ligand on brain endothelial health during ischemic injury has not been investigated. Therefore, the objective of this study was to determine human and murine brain‐derived endothelial cell S1PR1 expression profiles in response to in vitro ischemic‐like conditions or in vivo ischemia respectively. In adult C57BL/6blk male and female mice, S1PR1 expression was examined in cell suspensions from whole brain 24h post 60‐min tMCAO via flow cytometric (BD FACSARIA) analysis. Quantitative expression was determined in gated cell sub‐types using an anti‐mouse S1PR1/EDG‐1 conjugated fluorescent antibody. S1PR1 expression levels were also examined in primary male human brain microvascular endothelial cells (HBMEC) at passage 7 using standard immunoblotting. Additionally, the impact of RP101075 (novel selective S1PR1 ligand) on cyclooxygenase‐2 (COX‐2) levels was assessed following hypoxia plus glucose deprivation (HGD; 3h at 1% O2) or normoxia (21% O2). Brightfield microscopy was used to visualize HBMEC density and morphology, and proliferation was determined via bromodeoxyuridine and flow cytometric analysis. Basal levels of S1PR1 expression were detected in both human and murine brain‐derived endothelial cells. HGD and tMCAO resulted in increased endothelial S1PR1 expression levels. RP101075 partly preserved HGD‐induced morphological changes in HBMEC and elicited a trend in attenuating HGD‐induced HBMEC viability. Equally importantly, RP10175 attenuated HGD‐induced increases in COX‐2 levels, a recognized marker of ischemia‐induced vascular inflammation. In conclusion, selective S1PR1 ligands may serve as important beneficial mediators against ischemic damage in the brain endothelium through S1PR1 receptor upregulation and by attenuating brain endothelial cell inflammation and dysfunction.Support or Funding InformationUniversity of Arizona Valley Research Partnership Grant VRP37 P2 (RJG, FS).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Combined Treatment of Cyclosporine and Mycophenolate Improves Endothelial Function in Aorta of LDLR−/− Mice
Authors: Meng Mei , Shi Yi 
Published: June 2021

BackgroundChronic inflammation is a key player in the process of atherosclerosis. The present study was designed to investigate effects of immunosuppressive agents on endothelial dysfunction in a mouse model of atherosclerosis.MethodsTo induce atherosclerosis, 6‐week old LDLR knockout mice were fed with western diet for two months. Clinically used immunosuppressive agents, cyclosporine and mycophenolate, were mixed in chow. Aortic rings were fixed in the organ chambers for isometric tension measurement. Atherosclerotic plaques in aortic arch were visualized using Oil red O (ORO) staining and the level of CD68, COX‐2 positive cells was studied by Immunofluorescence. Lipoprotein and creatinine levels was measured using serum.Key resultsORO staining showed that atherosclerosis plaques in the aortic arch were reduced in mice fed with immunosuppressive drugs, compared with those fed with western diet. Acetylcholine induced relaxations at low concentrations, but contractions at high concentrations in the aorta of LDLR knockout mice. The acetylcholine‐induced contractions, but not the relaxations, were significantly reduced in mice treated with immunosuppressive agents. In the presence of indomethacin (non‐selective inhibitor of cyclooxygenase), acetylcholine‐induced contractions were restored to relaxations in both groups. The CD68, COX‐2 positive cells in the plaques of the two groups were not significantly different. In addition, there was no significant difference in serum lipoprotein and creatinine levels between the two groups of mice.ConclusionImmunosuppressive treatment reduces atherosclerotic plaque and prevents endothelial dysfunction in LDLR knockout mice. The underlying mechanisms may involve cyclooxygenase pathway; further studies are warranted to confirm, or not, the postulation.Support or Funding InformationThe present study was supported by the National Nature Science Foundation of China (81573418) and ZSYXRC‐014This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lipoic Acid Prevents Renal Dysfunction in Experimental Diabetes
Authors: Fonteles Manassés Claudino , Silva Ariana Gomes , Fonseca Maria Júlia Sousa , Santos Cláudia Ferreira , lopes Davi Elisiário Lima , Silva Matheus Carlos Matos , Nascimento Nilberto Robson Falcão 
Published: June 2021

IntroductionDiabetic nephropathy affects approximately 30–40% of patients with type 1 diabetes mellitus and 8% to 10% of patients with type 2 diabetes, and is the main cause of end‐stage renal disease, requiring hemodialysis or renal transplantation.ObjectiveThe study evaluated the effect of lipoic acid for 60 days in the prevention of experimental diabetic nephropathy in diabetic rats. Male Wistar rats were divided into three groups: euglycemic (eugly; n = 12), untreated diabetic (Diab; n = 6), and diabetic treated with lipoic acid (AL) 20mg/kg every 12 hours Orally (Diab + AL; n = 12) for 60 days. The animals were induced to diabetes with intraperitoneal injection of streptozotocin (STZ) (65 mg/kg, i.p), diluted in citrate buffer (pH 4.5). The control group received only citrate buffer (0.1mL/100g; i.p). Renal function was evaluated in metabolic cages on days 0, 15, 30, 45 and 60, with collection of urine and blood. After 60 days, all the animals were assigned to functional studies of renal hemodynamics and pressure natriuresis, in addition to molecular marker studies of diabetic nephropathy.ResultsThe animals studied presented classic diabetes characteristics such as hyperglycemia, maintained throughout the experimental protocol, polyuria, polydipsia and glycosuria, whose treatment with lipoic acid, was able to improve all these alterations. After 60 days, they presented albuminuria, which was attenuated with AL treatment. The excreted fraction of sodium, was increased 4‐fold in the Diab group and was attenuated in the AL‐treated group. The natriuresis increased about 7‐fold in the Eugli group during the period of increased renal perfusion pressure, however, it was impaired by 65% in the control non‐treated diabetic animals. The AL treated group, however, had normal pressure natriuresis. The renal concentration of gluthatione (GSH) decreased by 48% in the diabetic group when compared to the Eugli group, and the AL treatment restored the GSH levels to control values. The lipoperoxidation marker, malondialdehyde, was increased in the Diab group (68%) but was normal in the AL‐treated diabetic group.ConclusionTreatment with lipoic acid is able to prevent the great majority of changes caused during the course of experimental diabetes, and could be an alternative to the composition of a primary prevention treatment in conjunction with metabolic control drugs.Support or Funding InformationFUNCAP, CAPES and CNPqThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TRPM8 channel activation triggers relaxation of pudendal artery and corpus cavernosum with increased vascular sensitivity in spontaneously hypertensive rats: Is it a new target for erectile dysfunction?
Authors: Silva Darizy Flavia , Wenceslau Camilla Ferreira , Mccarthy Cameron G , Szasz Theodora , Ogbi Safia , Webb Robert C 
Published: June 2021

Background and PurposeErectile dysfunction (ED) is frequently encountered in patients with arterial hypertension and there is a recently uncovered functional correlation between the expression of thermoreceptor channels TRPM8 (melastatin 8) and alterations in blood pressure. The aim of this study was to investigate the function of cold‐sensing TRPM8 channel in internal pudendal artery (IPA) in normotensive and hypertensive rats.Experimental ApproachWe performed experiments integrating physiological, pharmacological, biochemical, and cellular techniques.Key ResultsTRPM8 channels are expressed in the IPA and in vascular smooth muscle cells from IPA and corpus cavernosum (CC). In addition, TRPM8 activation, by both a cooling compound (icilin) and cold (thermal stimulus), induced relaxation in IPA and CC. In IPA, the concentration‐response curve to icilin was desensitized in different conditions, such as: the absence of vascular endothelium, the presence of L‐NAME (10−4 M) or indomethacin (10−5 M), a combination of charybdotoxin (10−7 M) and apamin (5×10−6 M), and Y27632 (10−6 M). Interestingly, icilin‐induced vasodilation was significantly higher in IPA from spontaneously hypertensive (SHR) compared to Wistar rats, despite no differences in the TRPM8 expression in IPA between the strains, suggesting that the sensitivity of TRPM8 channels is higher in SHR. In CC, the concentration‐response curve to icilin was shifted to the right in the presence of ROCK inhibitor Y27632, suggesting that ROCK, at least in part, is important to relaxation induced by TRPM8 activation. Supporting these functional data, we observed that the expression of RhoA and ROCK was not changed in CC treated with either vehicle, Phe, or Phe+icilin. However, MYPT1 phosphorylation was increased by Phe compared to control, and Phe+icilin reverted this effect, suggesting that TRPM8 activation inhibits RhoA/ROCK pathway in CC.ConclusionsThese data demonstrate, for the first time, the expression and function of TRPM8 channels in the IPA and CC involving, at least in part, endothelium‐derived relaxing factors and ROCK inhibition. Overall, this channel could potentially be a new target for the treatment of hypertension associated‐ED.Support or Funding InformationNational Council for Scientific and Technological Development (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES); NIDDK Diacomp Pilot & Feasibly Program.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lactobacillus rhamnosus GG supernatant prevents acute alcohol‐induced liver steatosis and injury through ER stress and autophagy‐mediated signaling pathways
Authors: Zhang Lan , Li Huimin , Kong Xiaoxia , Feng Wenke 
Published: June 2021

Acute alcohol exposure induces ER stress and autophagy in mouse liver, which protect alcohol‐induced cell death by removing damaged proteins and cellular compartments. We have previously demonstrated that Lactobacillus rhamnosus GG culture supernatant (LGGs) prevents acute alcohol‐induced hepatic steatosis and injury. The purpose of this study was to determine whether the protective effect of LGGs against acute alcohol‐induced liver injury through regulating autophagy and ER stress signaling pathways. C57BL/6 mice on standard chow diet were supplemented with supernatant from LGG culture (109 colony‐forming unit/mouse) for 5 days, and one dose of alcohol at 6 g/kg body weight was administered via gavage. Liver tissues and serum samples were collected 6 hours after alcohol administration. LGGs pretreatment significantly suppressed acute alcohol‐induced liver injury measured by serum activities of alanine aminotransferase (ALT) and asparagine aminotransferase (AST), and reduced liver steatosis assessed by triglyceride content and Oil red O staining of the liver tissues. Ethanol exposure increased LC‐3II/LC‐3I protein ratio suggesting an upregulated autophagy response, which was normalized by LGGs pretreatment. mRNA levels of molecules involved in autophagy formation and mitophagy regulation including beclin‐1, Atg7, Atg5 and p62 were significantly increased in alcohol‐treated mouse livers and suppressed by LGGs pretreatment. Alcohol‐induced ER stress is a trigger of autophagy regulation. Our studies also showed that LGGs pretreatment decreased acute alcohol‐induced hepatic GRP78 and CHOP expression, indicating a down‐regulation of ER stress by LGGs. Further studies showed that LGGs significantly reduced alcohol‐induced ROS levels assessed by DHE staining and Cyp2e1 and NRF2 protein levels in the liver samples. These results suggest that LGGs prevent the acute alcohol‐induced liver injury by preventing ROS production leading to a decreased ER stress and a normalization of hepatic autophagy response to acute ethanol exposure. The use of bacteria‐free LGG culture supernatant provides a novel strategy for prevention of acute alcohol‐induced liver injury (LZ and HL contributed equally to this study).Support or Funding InformationNIHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TCV116 Ameliorates Asthma Features in Murine Allergic Asthma Model
Authors: Son So‐Eun , Kim Mi‐Jeong , Im Dong‐Soon 
Published: June 2021

BackgroundSeveral studies have reported on the expression of the type I angiotensin II receptor in human lung tissues. In severe acute attacks of asthma, the renin‐angiotensin system is activated by elevated plasma concentrations of renin and angiotensin II. Furthermore, angiotensin II potentiates methacholine‐induced bronchoconstriction in patients with mild asthma. However, the in vivo anti‐allergic effect of type I angiotensin II receptor antagonists has been poorly evaluated. We aimed to investigate effect of TCV116, an antagonist of the type I angiotensin II receptor on allergic asthma.METHODSTo elucidate whether TCV116 lactone has beneficial effects on allergic asthma, anti‐asthma effects were studied using female Balb/c mice and rat RBL‐2H3 mast cells. Antigeninduced degranulation was measured in vitro by measuring b‐hexosaminidase activity. A murine ovalbumin‐induced allergic asthma model was used to test the in vivo efficacy of TCV116.RESULTSIt was observed that while TCV116 inhibited the antigen‐induced degranulation in rat RBL‐2H3 mast cells. Administration of TCV116 and irbesartan decreased the number of immune cells in the bronchoalveolar lavage fluid and reduced the expression of Th2 (IL‐4, IL‐5, and IL‐13) and Th1 cytokines (IL‐2 and IFN‐g) in the lung tissues of mice with ovalbumin‐induced allergic asthma. Histological studies revealed that TCV116 and irbesartan reduced inflammation and mucin production in the lungs.CONCLUSIONThe present findings provide evidence that TCV116 and irbesartan could have potential applications as anti‐allergic agents.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MicroRNA 7 Regulates YAP/TAZ Signaling in Pulmonary Vascular Stiffness and Pulmonary Hypertension
Authors: Woodcock Chen‐Shan , Hafeez Neha , Estephan Leonard , Speyer Gil , Handen Adam , Kim Seungchan , Bertero Thomas , Chan Stephen 
Published: June 2021

Pulmonary hypertension (PH) refers to a set of heterogeneous vascular diseases defined by sustained elevation of pulmonary arterial pressure (PAP) and pulmonary vascular resistance (PVR) leading to right ventricular (RV) remodeling and subsequent death. In particular, early increased pulmonary artery stiffness drives vascular remodeling during PH development. Specifically, our group has demonstrated that two transcriptional coactivators involved in the Hippo pathway, Yes‐associated protein 1 (YAP1) and TAZ (or WWRT1), play a critical role in pulmonary vascular cell survival and proliferation in response to mechanoactivation by extracellular matrix (ECM) stiffness. This study sought to identify novel molecular regulators driven by pulmonary vascular stiffness in the context of YAP1/TAZ signaling in PH. As determined by RNA sequencing and confirmed by RT‐qPCR, in primary pulmonary arterial endothelial cells (PAECs) cultured on soft (0.5 kPa) or stiff (50 kPa) matrix, stiff ECM increased YAP1/TAZ activation and correspondingly decreased the expression of microRNA‐7 (miR‐7). Conversely, forced expression of miR‐7 down‐regulated expression of both YAP1 and TAZ. The biogenesis of miR‐7 has been suggested to be controlled by the RNA binding protein QKI. Correspondingly, in PAECs cultured in stiff ECM, QKI expression was increased; suppression of QKI under these conditions significantly increased miR‐7 expression. Finally, forced miR‐7 expression induced apoptotic caspase 3/7 activity in PAECs, and this observation was consistent to gene set enrichment analysis (GSEA) of RNA sequencing data, which suggested miR‐7 might modulate the apoptotic pathway. Correspondingly, These observations suggest that miR‐7 may serve as a critical molecule that regulates YAP1/TAZ signaling and downstream PAEC pathophenotypes in pulmonary vascular stiffness relevant to PH.Support or Funding InformationThis work was supported by R01 HL124021, HL 122596, HL 138437, and UH2 TR002073 as well as the AHA grant 18EIA33900027 (S.Y.C.)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Heterocellular contact can dictate arterial function
Authors: Ruddiman Claire A. , Shu Xiaohong H. , Keller T.C. Stevenson , Keller Alexander S. , Yang Yang , Good Miranda E. , Best Angela K. , Columbus Linda , Isakson Brant E. 
Published: June 2021

Resistance arteries use different dilatory heterocellular signaling mechanisms than conduit arteries (endothelial derived hyperpolarization versus nitric oxide). Anatomically, resistance arteries use myoendothelial junctions (MEJs), endothelial cell (EC) extensions that make contact with smooth muscle cells (SMCs), which are not present in conduit arteries. The objective of this study was to determine if the presence of MEJs in conduit arteries can alter heterocellular signaling. We previously demonstrated that plasminogen activator inhibitor‐1 (PAI‐1) can regulate formation of MEJs. Thus, we applied pluronic gel containing PAI‐1 directly to conduit arteries (carotid arteries, CAs) in live mice to determine if this could induce formation of MEJs. We found a significant increase in EC projections that resemble MEJs, correlating with increased biocytin dye transfer from ECs to SMCs and dilation to NS309. Next, we used pressure myography to investigate whether these structural changes were accompanied by a functional change in vasodilatory signaling. Interestingly, PAI‐1‐treated CAs underwent a switch from conduit to resistance artery vasodilatory profile via increased endothelial‐derived hyperpolarization (EDH) signaling and diminished nitric oxide (NO) signaling. Following PAI‐1 application, we also found a significant increase in carotid expression of endothelial alpha globin, a protein predominantly expressed in resistance arteries. Carotids from mice lacking alpha globin (Hba1−/−) demonstrated that L‐NAME, an inhibitor of NO signaling, was able to prevent vasodilation. Thus, the presence or absence of MEJs is an important determinant for protein expression to influence heterocellular communication. In particular, our data suggests that alpha globin may allow for EDH to predominate in resistance arteries.Support or Funding InformationThis work was supported by HL088554 (BEI) and the National Natural Science Foundation of China 81672945 (XHS).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.