bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anti‐inflammatory effect of miR‐17‐3p in human umbilical vein endothelial cells (840.16)
Authors: Zhang Yu , Leung Susan , Man Ricky 
Published: June 2021

Anti‐inflammatory effect of miR‐17‐3p in human umbilical vein endothelial cells Yu ZHANG, Susan WS LEUNG and Ricky YK MAN Department of Pharmacology &amp; Pharmacy, the University of Hong Kong, Hong Kong SAR, China Aim: MicroRNAs (miRNAs) are a class of small, noncoding RNAs of ≍22 nucleotides that negatively regulate gene expression. They appear to play a role in the development/progression of many disorders, including inflammation, cardiovascular diseases and endothelial dysfunction. This study examines the role of miR‐17‐3p in vascular inflammation.Methods: Human umbilical vein endothelial cells (HUVECs) were transfected with miR‐17‐3p agomir (miR‐17‐3p mimic) or its negative control using lipofectamine 2000. They were incubated with lipopolysaccharide (LPS, 10 ng/ml) for 16 hours to induce inflammatory reactions. The level of miR‐17‐3p and the expressions of IκBα, p65 and phosphorylated p65 (p‐p65) were measured by qPCR and Western blotting. The amount of interleukin‐8 (IL‐8) and tumor necrosis factor (TNF)‐α released in the culture medium was detected with ELISA kit.Results: The levels of miR‐17‐3p, IL‐8, TNF‐α and p‐p65 were increased following LPS stimulation. While the expression of p65 was not changed, the expression of IκBα was reduced. LPS‐induced increase in IL‐8 level and decrease in IκBα expression were smaller in HUVECs transfected with miR‐17‐3p agomir than in those transfected with the negative control.Conclusion: MiR‐17‐3p is up‐regulated in LPS‐induced inflammation. It may produce an anti‐inflammatory effect by inhibiting NF‐κB pathway and reducing IL‐8 production.
Contribution of Rho kinase to TRPM4‐mediated myogenic tone in cerebral parenchymal arterioles (841.5)
Authors: Li Yao , Baylie Rachael , Tavares Matthew , Brayden Joseph 
Published: June 2021

Cerebral parenchymal arterioles (PAs) play a critical role in assuring appropriate blood flow and perfusion pressure within the brain. PAs are unique in contrast to cerebral pial arteries, as defined by their critical roles in neurovascular coupling and distinct sensitivities to mechanical and chemical stimulants. Elevation in intraluminal pressure, a physiologic vasomotor stimulant, causes smooth muscle membrane depolarization and contraction (myogenic tone) in cerebral arteries. However, the mechanisms by which this myogenic tone is regulated in the cerebral microcirculation remain unclear. Prior research in our laboratory indicates that myogenic tone of PAs is mediated primarily through stimulation of mechano‐sensitive P2Y purinergic receptors. The objective of the present study was to determine the roles of TRPM4 channels and Rho kinase signaling in PA myogenic tone. We show that suppression of TRPM4 channel expression in vivo using antisense oligonucleotides reduced (by ~40%) both myogenic and P2Y‐receptor agonist‐induced tone in PAs. Further, we found that the Rho kinase inhibitor H1152 reversibly and nearly completely inhibited myogenic tone in these brain arterioles (IC50:0.2 µM). H1152 (1 µM) also attenuated P2Y4 and P2Y6 receptor‐mediated vasoconstriction by 85% and 87%, respectively. Finally, H1152 (1 μM) reduced constitutive and UTPγS (0.5 μM)‐activated TRPM4 currents in parenchymal myocytes by 61% and 75%, respectively. These results indicate an important role for Rho kinase in regulation of TRPM4 channel activity and myogenic tone in the cerebral microcirculation.Grant Funding Source: Supported by NIH HL095488
Glyoxalase‐1 gene transfer prevents cerebral deficits in rats with type 1 diabetes (841.9)
Authors: Alomar Fadhel , Shao Chun , Xia Jianxun , Xiong Huangui , Mayhan William , Bidasee Keshore 
Published: June 2021

Background: Individuals with type 1 diabetes (T1D) confront daily challenges including reduction in sustained attention, decreased decision‐making abilities and increased risks for accidents. Studies attribute these insufficiencies in part to reduced psychomotor efficiency, slowing in information processing speed, impaired visuo‐construction and cerebral vascular dysfunction. To date, the molecular cues responsible for these cerebral deficits in T1D remain poorly defined. Here we investigated whether the reactive carbonyl species methylglyoxal (MGO), whose production is upregulated in T1D, is an underlying cause.Methods and results: Brain levels of MGO were 6 times higher in T1D compared to non‐diabetic rats. Chronic administration of MGO to control rats impaired their nest building capabilities, an activity that requires decision‐making, visual and motor functions. MGO treatment also impaired endothelial‐mediated vasodilatation of cerebral microvessels, induced localized microvascular leakages, astrocytes activation and impaired synaptic transmission, akin to that seen in T1D. Increasing expression of the MGO‐degrading enzyme glyoxalase‐1 (Glo‐1) in rats after the onset of diabetes using an adeno‐associated viral construct, decreased brain MGO level and blunted cerebral impairments induced by T1D.Conclusion: From these data we conclude that upregulation of MGO is an underlying cause for cerebral deficits in T1D and MGO detoxification is sufficient to prevent these cerebral dysfunctions.
Inhibitory effect of AHCC and GCP on tumorigenesis of C6 glioma cells in culture medium and rats mediated by downregulating CD133 and Nestin expressions (842.13)
Authors: Zhang Xiao‐Hong , Nishioka Hiroshi , Fujii Hajime 
Published: June 2021

The CD133 and Nestin proteins are described as cancer stem cell markers. Norepinephrine (NE) is a sympathetic neurotransmitter, studies have indicated the level of NE or the active degree of adrenoceptors in certain bodily tissues correlates with the degree of tumorigenesis in those tissues. To explore the relation between the activation of adrenoceptors and the expression level of CD133 and Nestin proteins, we investigated whether NE might regulate the expression of CD133 and Nestin and its signaling pathway; the effect of AHCC (Active Hexose Correlated Compound) &and GCP (Genistein Concentrated Polysaccharide) on the expression of CD133 and Nestin and the sphere‐colony formation of C6 cells in medium and the tumorigenesis in rats inoculated subcutaneously with C6 cells.We measured the expression level of CD133 and Nestin in C6 cells with western blot analysis, examined the sphere‐colony formation of the cells by sphere assay and measured the tumor volume in rats calculated as length × width × depth × 0.5236.We found out the expressions of CD133 and Nestin in C6 cells were significantly upregulated by NE stimulation. Further analysis showed prazosin completely blocked the NE‐induced upregulation of CD133 and Nestin expressions, while propranolol had no influence to them. Pretreatment with LY294002, blocked the NE‐induced upregulation of CD133 expression, but showed no influence to Nestin expression. Pretreatment with B3806, blocked the NE‐induced upregulation of Nestin expression, but had no significant influence to CD133 expression in the cells. Co‐culture with AHCC or GCP, the upregulated effects on the expressions of CD133 and Nestin induced by NE in the cells were weaken and the sphere size of the cells became smaller. The animal experiment in rats showed that the tumor size was markedly smaller in AHCC or GCP group than in control group.Based on these findings, we thus assume that AHCC & GCP can ameliorate some kinds of tumorigenesis in cancer stem cells expressing CD133 and Nestin proteins.
Silencing of regulator of G‐protein signaling 10 (RGS10) increases phosphorylation of eukaryotic initiation factor 4E binding protein (4E‐BP1), and enhances growth and survival signaling pathways in ovarian cancer (843.11)
Authors: Altman Molly , Patel Mihir , Hooks Shelley , Murph Mandi 
Published: June 2021

Ovarian cancer is a devastating disease that is the 5th leading cause of cancer related deaths in women in the US. It is difficult to treat in that more than 70% of patients that are initially responsive to first‐line chemotherapeutics become refractory in less than 2 years. The problem of drug‐resistance or chemoresistance has plagued cancer researchers and hindered the success of cancer therapy development. In our previous studies examining computational bioinformatics, we demonstrated that reduced RGS gene expression develops with the occurrence of drug resistance in ovarian cancer cell lines. Specifically, we have shown that silencing of RGS10 protein increases viability of ovarian cancer cells in the presence of cycle arrest compounds such as paclitaxel. In our present study, we have shown that silencing RGS10 in ovarian cancer cell lines increases phosphorylation of Eukaryotic initiation factor (eLF) 4E, the mRNA 5’ cap‐binding protein (phosphorylated‐4E‐BP1). Phosphorylation of 4E‐BP1 has been shown to be a hallmark of more aggressive cancer phenotypes in prostate cancer and ovarian cancer. HeyA8 Parental and SKOV3 ovarian cancer cell lines were transiently transfected with siRNA non‐targeted control and human RGS10. Approximately 36 hrs post‐transfection cells were serum‐starved and treated with INK128 a selective mTOR inhibitor overnight. Cells were treated the following day with lysophosphatidic acid (LPA) and collected for western blot analysis. The same treatment conditions were also analyzed by immunofluorescence assays and imaged using a Cellomics ArrayScan VTI reader and HCS software. We found that when RGS10 was silenced in these ovarian cancer cell lines we observed changes in cell growth and size. Based on the results from these assays, we believe that we have elucidated a novel role of RGS10 in the growth and survival signaling pathways involved in the development of chemoresistance in ovarian cancer. Grant Funding Source: NIH
Intragenic suppressors of McCune‐Albright syndrome R201H mutations also suppress other constitutively active Gs alleles (843.5)
Authors: Rylaarsdam Robin , Turcic K , Dafour A , Carroll J 
Published: June 2021

McCune‐Albright Syndrome (MAS) is a genetic disorder caused by a mutation in Gs alpha at Arg201 that inhibits GTP hydrolysis, constitutively activating the protein. We identified three sites that, when mutated, suppressed the constitutive activity caused by R201H: F142, R231, and L266. None of the three mutations by themselves caused constitutive activity of the protein or significantly changed cellular responsiveness to hormone, as measured by the ability of a luteinizing hormone receptor agonist to activate cAMP production. We have now examined the ability of mutations at these three sites to suppress other activating mutations of Gs alpha. Other McCune‐Albright associated mutations include R201C and R201S. Both F142S and R231C were able to suppress the constitutive activity of both of previously untested R201 substitutions. L266N was able to suppress R201S, and experiments investigating its effects on R201C are ongoing. Gs alpha can also be constitutively activated by the Q227L substitution, associated with a variety of adenomas. The mutations identified as suppressors of R201H were all unable to suppress the constitutive activity of Q227L, suggesting that the action of the suppressors is specific to the R201 site and not generalizable to other activating perturbations of Gs alpha.Grant Funding Source: Supported by NIH grant 1R15ED020190‐01
Acrolein exacerbates anti‐retroviral therapy‐associated hepatotoxicity by increasing ER stress, apoptotic gene expression and cell death (844.14)
Authors: Ghare Smita , Donde Hridgandh , Joshi‐Barve Swati , McClain Craig , Barve Shirish 
Published: June 2021

In HIV patients prolonged ART usage accompanied by other life style components (e.g. alcohol consumption, smoking) has been associated with adverse hepatic events caused by abnormal lipid accumulation and peroxidation. However, role of cofactors that can significantly alter the hepatic interaction of ART leading to pathogenic outcomes is only poorly understood. In this regard, ART mediated hepatic steatosis can give rise to highly reactive hepatotoxic aldehydes, like acrolein, via lipid peroxidation. Hence the present study examined the role of acrolein as a cofactor in ART drug, azidothymidine (AZT)‐induced hepatotoxicity.Cytotoxic effects of acrolein and AZT were investigated using human hepatoma cells (HepG2) and rat‐primary cells. Cells were treated with various concentrations of acrolein (ACR) and AZT alone or in combination (AZT+ACR). Our results showed that acrolein or AZT alone had minimal effect on hepatocyte survival. However combinatorial treatment of AZT+ACR significantly increased apoptotic death. Further, the hepatotoxic effects of AZT+ACR were observed to involve ER stress and mitochondrial dysfunction which was documented by Ca 2+ release, up‐regulation in ER stress genes and mitochondrial membrane depolarization and ATP depletion. Additionally, the combinatorial treatment increased apoptotic gene expression of FasL, a known contributor to liver damage. Notably, acute acrolein exposure enhanced AZT responsive epigenetic modifications at the FasL promoter contributing to transcriptional activation. Overall, the data suggest that presence of acrolein plays a significant pathogenic role in the development of ART‐induced hepatotoxicity.Grant Funding Source: Supported by NIH and Veterans Administration
Study the effect of drug resident time on in vivo efficacy (844.6)
Authors: Lee Kin Sing Stephen , Yang Jun , Ng Connie , Niu Jun , Morisseau Christophe , Hammock Bruce 
Published: June 2021

When developing a new drug, it is difficult to predict from in vitro parameters to in vivo efficacy. It is generally accepted that drugs with the strong interactions with their target are more likely to show better efficacy. Therefore, thermodynamic parameter: dissociation constant or inhibition constant (Ki) are often considered as the most important factor to be determined as the first screening to prioritize the drug candidates. However, the kinetic parameter: the dissociation rate constant (koff) of the drugs is often neglected although several reports suggested that koff is as important as or more than Ki to estimate in vivo efficacy.Soluble epoxide hydrolase (sEH) has been proven to play an important role on regulating important lipid mediators, epoxy‐polyunsaturated fatty acid. Inhibition of sEH has been showed to be beneficial to human health in several aspects, anti‐inflammatory, analgesic and promoting tissue regeneration. We recently synthesized a series of new sEH inhibitors with different Kis and koffs. These inhibitors will be used as tools to test if koff can predict in vivo efficacy as equal as or better than the Ki.Here, a series of inhibitors with improved in vitro efficacy will be reported. Our in vivo results suggested that koff is crucial parameters to affect drug efficacy in an alternate way.Grant Funding Source: National Institute of Environ‐ mental Health Sciences (NIEHS) grant ES002710
Targeting hyperactive Rho GTPase regulatory proteins in autism (845.7)
Authors: Martinez Luis , Tejada‐Simon Maria 
Published: June 2021

Cognitive impairment in neurological disorders like Fragile X syndrome (FX), a leading cause of autism, is thought to be due to abnormal neuroplasticity. These learning‐induced modifications require reorganization of the actin cytoskeleton within dendritic spines, a dynamic process that is known to be regulated by the Rho family of small GTPases. In a mouse model of FX we found increased levels of the Rho GTPase Rac1 in the hippocampus, a brain structure involved in learning. The FX mouse hippocampus also exhibits impaired synaptic plasticity and malformed dendritic spines. In an effort to define whether there is a critical role for the small GTPases in FX‐associated learning deficiencies, Rac1 was pharmacologically targeted using novel agents that inhibit its activation and therefore, its function. Mice treated chronically with Rac1 inhibitors were subjected to behavioral tasks that examine contextual and cued memory (associated fear conditioning) as well as spatial memory (Morris Water maze). Performance on these tasks is dependent on the neuronal integrity of the hippocampus. Our results showed that FX mice treated with the inhibitors improved their cognitive capabilities. These findings suggest that Rac1 hyperactivity may play an important, yet reversible role in FX and autism‐related molecular pathology contributing to cognitive difficulties.Grant Funding Source: Supported by the Jérôme LeJeune Foundation (France), FRAXA Research Foundation (USA) and the Grants
Biologically‐inspired ApoE3‐reconstituted high density lipoprotein: toward development of nanomedicines for alzheimer's disease (846.2)
Authors: Qingxiang Song , Xiaoling Gao , Meng Huang 
Published: June 2021

Amyloid beta (Aβ) accumulation in the brain plays a central role in Alzheimer’s disease (AD) pathogenesis, and the common late‐onset form of AD is characterized by an overall impairment in Aβ clearance but not in Aβ production. Therefore, development of nanostructures that can facilitate Aβ clearance represents a promising strategy for AD intervention. Here, we hypothesized that biologically‐inspired nanostructure_ApoE3‐reconstituted high density lipoprotein (HDL) (ApoE3‐rHDL) which presents high binding affinity to Aβ might serve as a novel nanomedicine for disease modification in AD by accelerating its clearance. Surface plasmon resonance, transmission electron microscopy and co‐immunoprecipitation analysis showed that ApoE3‐rHDL demonstrated high binding affinity to both Aβ monomer and oligomer. ApoE3‐rHDL also accelerated the microglial, astroglial and liver cell degradation of Aβ by facilitating the lysosome transport pathway. One hour after intravenous administration, about 0.4% ID/g of ApoE3‐rHDL got access to the central nervous system. Four‐week dosing of ApoE3‐rHDL decreased amyloid deposition, attenuated microgliosis, ameliorated neurologic changes and rescued memory deficits in an AD animal model. The findings here indicated that biologically‐inspired nanostructure ApoE3‐rHDL could serve as a novel nanomedicine for disease modification in AD by accelerating its clearance, which also justified the concept that nanostructures with Aβ‐binding affinity might provide a novel nanoplatform for the treatment of AD.Grant Funding Source: This study was supported by National Natural Science Foundation of China (No. 81373351), National K
The selective BACE1 inhibitor VIa reduces Aβ production in a mouse model of Alzheimer’s disease (846.4)
Authors: Cheng Xiao‐rui , Zhou Yu , Gu Wei , Nie Ai‐hua , Cheng Jun‐ping , Zhou Wen‐xia , Zhang Yong‐xiang 
Published: June 2021

We are pursuing potentβ‐site amyloid precursor protein cleaving enzyme 1 (BACE1) inhibitors in an effort to identify suitable Alzheimer’s disease (AD) drug candidates. Employing a homogeneous, time‐resolved fluorescence technique, we determined the selectivity of compound VIa. We then conducted a cell‐based assay using the huAPPswe/huBACE1 transgenic CHO cell to investigate the effect on the β‐cleavage of APP. The effect of VIa anti‐AD was valued using both acute and subchronic dosing paradigms in PrPhAPPswe/PS1ΔE9 transgenic mice. Our results have shown that the novel compound VIa exhibits potent inhibitory effects with and IC50=5.9 nM and displays 30.8‐fold, 7500‐fold and 17533‐fold selectivity against the other aspartic proteases BACE2, cathepsin D and renin, respectively. In cellular assays, VIa moderately reduces Aβ production: Aβ1‐40 with an IC50 =143 nM and 1 nM VIa reduced Aβ1‐42 by 40.17%. Concomitant with VIa inhibiting the β‐cleavage of amyloid precursor protein (APP), VIa increases the production of sAPPα with an approximate EC50 of 16.5 nM. In testing this compound’s efficacy in vivo, the oral administration of VIa resulted in a significant decrease in Aβ1‐40 and Aβ1‐42 in the blood of a mouse model of AD by 17.5‐72.44% and 14.5‐80.32%, respectively. This result indicates that the novel compound VIa is a small, potent, selective, and nonpeptidic BACE1 inhibitor.Grant Funding Source: the National Science and Technology Major Project (2012ZX09301003‐002‐001, 2013ZX09508104)
Liver X receptors activation decreases neuroinflammation in Parkinson’s disease (846.5)
Authors: Campolo Michela , Paterniti Irene , Siracusa Rosalba , Bruschetta Giuseppe , Cuzzocrea Salvatore , Esposito Emanuela 
Published: June 2021

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the degeneration of dopaminergic nigrostriatal neurons, progressive loss of substantia nigra and inflammation. Liver X receptors (LXRs) are considered as potent anti‐inflammatory molecules in macrophages and neuronal cells. The aim of this study was to investigate the role of LXRs in neurodegenerative disorders like PD using in vivo, in vitro and ex vivo models; for this purpose, we studied the effect of the synthetic LXR agonist, TO901317, in neuroinflammatory pathway related to PD.Our results showed a reduction in neurotoxin 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP)‐induced cell death, iNOS and COX2 expression, IκB‐α degradation, and NF‐κB translocation after treatment with TO901317. Moreover, in in vitro study, through SH‐SY5Y cells, we demonstrated an important restoration of neuthrophic factors (GDNF) and nNOS expression.Moreover, in in vivo study, mice exhibited a significantly recovery of motor function after TO901317 treatment.In the light of the results obtained, TO901317, a synthetic LXR agonist, is able to modulate the neuroinflammatory pathway involved in PD increasing the locomotors function. Therefore, LXRs could be considered as a possible therapeutic target in a neurodegenerative disorders like PD.
Properties of the high‐conductance Ca2+‐activated K+ (BK) channel in pulmonary arterial smooth muscle cells (847.3)
Authors: Detweiler Neil , Song Li , Versluis Rachel , Nelson Piper , Rhee Sung , Rusch Nancy 
Published: June 2021

We reported an upregulation of an oxygen‐insensitive splice variant of the high‐conductance Ca2+‐activated K+ (BK) channel in pulmonary arteries (PA) from rats with chronic hypoxia (CH)‐induced pulmonary hypertension (PH). This finding infers that the BK channel may represent a vasodilator target for the treatment of PH. However, the BK channel in pulmonary arterial smooth muscle cells (PASMCs) reportedly shows low Ca2+‐sensitivity due to uncoupling of the α‐subunit pore from the requisite accessory β1 subunit. The goal of this study was to: i) define the Ca2+‐sensitivity of the BK channel in rat PASMC, and ii) evaluate if a BK channel opener that relies on the β1 subunit activates BK channels in PASMCs. Whole‐cell K+ current in patch‐clamped PASMCs from normoxic (N) and CH rats failed to reveal BK current, although it was observed under the same conditions in cerebral ASMCs. However, the Ca2+‐sensitivity of BK channels in inside‐out patches from PASMCs and cerebral ASMCs exposed to a range of free Ca2+ concentrations (10‐7 to 10‐4 mol/L) was not significantly different. Additionally, BK channel open probability (NPo) was increased 19‐fold by lithocholate (45 μmol/L), a BK channel activator that requires the β1 subunit. Thus, the BK channel in PASMCs may be unique in some aspects, but it appears to show a normal level of Ca2+‐sensitivity and be coupled to a functional β1 subunit.Grant Funding Source: Supported by AHA 13PRE17240055 (NDD), NIH UL1RR029884 (NDD), gift from the Dean’s Society (UAMS COM)