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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 739 records · Page 41
All sourcesNASA NTRSDOE OSTI
DOE OSTI2018

The gene expression of two endoplasmic reticulum aminopeptidase 1 isoforms is regulated by distinct posttranscriptional mechanisms

Highlights: • ERAP1 contains two isoforms that exchange 3′ untranslated regions of mRNA. • One of ERAP1 isoform is suppressed the protein expression posttranscriptionally. • We identified cis-elements that repressed the alternative splicing within ERAP1. • IFN-γ stimulation affects transcription and the alternative splicing of ERAP1. Endoplasmic Reticulum Aminopeptidase 1 (ERAP1) is a multifunctional enzyme belonging to the M1 family of aminopeptidases and shown to be associated with various autoimmune diseases. Human ERAP1 protein has two isoforms produced by alternative splicing of the 3′ terminal exon, although their functional differences have not yet been fully clarified. In this study, we showed that the isoforms undergo different posttranscriptional regulation mechanisms via their respective 3′ untranslated regions. Using a reporter system, we identified several cis-elements that are important for the regulation of alternative splicing. Finally, we revealed a close relationship between the transcriptional induction of the ERAP1 gene by interferon-gamma and the alternative splicing. These results suggest that the two ERAP1 isoforms function under different pathophysiological conditions.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

MiR-590-5p alleviates intracerebral hemorrhage-induced brain injury through targeting Peli1 gene expression

Highlights: • Down-regulation of miR-590-5p in ICH mice and LPS-induced microglia cells. • MiR-590-5p attenuates ICH-induced brain injure. • Peli1-1 is a direct target of miR-590-5p. • Peli1 overexpression abrogates the effect of miR-590-5p mimic on ICH-induced brain injure. Intracerebral haemorrhage (ICH) is a common and devastating cerebrovascular disease with high morbidity and mortality, and its pathophysiological mechanisms were complex and still unclear. Increasing researches reported that microRNAs (miRNAs) played an important role in ICH-induced brain injury and microglial activation. In this study, we investigated the biological function of miR-590-5p and explored its molecular mechanism in ICH mice. The results of qRT-PCR showed that miR-590-5p expression level was down-regulated in perihematomal brain samples of ICH mice compared with that of sham group. In LPS-induced microglia cells, miR-590-5p level was also down-regulated at 24 h post-LPS compared with that of control group. Moreover, miR-590-5p overexpression remarkably increased the cerebral water content and neurological severity scores compared with that of scramble group in ICH mice. The production of inflammatory cytokine including IL-6, interleukin (IL)-1β, and tumor necrosis factor (TNF)-α in ICH mice was notably inhibited by miR-590-5p overexpression. Furthermore, the results of dual-luciferase reporter assay indicated that Pellino-1 (Peli1) was a direct target of miR-590-5p. MiR-590-5p overexpression remarkably inhibited the Peli1 gene expression both mRNA and protein levels. In addition, Peli1 overexpression partly abrogated the inhibitory effect of miR-590-5p mimic. Taken together, these datas suggested that miR-590-5p attenuated brain injury in ICH mice through inhibiting Peli1 gene expression, indicating that miR-590-5p may be a promising molecular target for ICH treatment.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

E3317 promotes cholesterol efflux in macrophage cells via enhancing ABCA1 expression

Highlights: • E3317 was first found as an ABCA1 upregulator in an ABCA1 transcriptional activity assay. • E3317 increased ABCA1 mRNA and protein expression in both L02 and RAW264.7 cells. • E3317 promoted cholesterol efflux and reduced cellular lipid accumulation in RAW264.7 macrophages dependent on ABCA1. • E3317 upregulated ABCA1 expression through activating PPARγ. • Y327 and F363 are the key PPARγ binding epitopes of E3317. Reverse cholesterol transport (RCT) plays an important role in cholesterol and lipid metabolism. Regulating the activities of key transporters and receptors in RCT, such as ATP-binding cassette transporter A1 (ABCA1), helps to prevent atherosclerotic cardiovascular disease. In this study, we used an ABCA1 promoter luciferase reporter assay to screen 20,000 compounds for ABCA1 upregulators. Compound E3317 (N-(6-butylbenzo[d]thiazol-2(3H)-ylidene)-3-(N-(2-cyanoethyl)sulfamoyl)benzamide)) was identified as a positive hit with an EC{sub 50} value of 0.2 μM in ABCA1p-LUC HepG2 cells. Thus, we hypothesized that E3317 might have cholesterol- and lipid metabolism-regulating effects through ABCA1 upregulation. E3317 significantly increased ABCA1 mRNA and protein expression in hepatic L02 cells and RAW264.7 macrophages. E3317 promoted cholesterol efflux to apolipoprotein A-I in RAW264.7 macrophages and significantly decreased lipid accumulation in oxidized low-density lipoprotein-induced murine RAW264.7 macrophages. Further studies using ABCA1 siRNA showed that the promotion of cholesterol efflux and decrease of lipid accumulation by E3317 depended on ABCA1 expression. Mechanistic studies indicated that E3317 regulated ABCA1 expression via activating nuclear receptor peroxisome proliferator-activated receptor gamma (PPARγ), which plays an important role in the regulation of glucose homeostasis and lipid metabolism. The structure of E3317 was docked in the ligand-binding domain of PPARγ (PBD code: 4EMA) to find the key binding amino acids. Site mutation assays confirmed that Y327 and F363 were the key PPARγ binding epitopes of E3317. Our results revealed that E3317 upregulates ABCA1 expression and thereby promotes cholesterol efflux. E3317 may regulate ABCA1 expression through PPARγ. Our findings provide a new compound, E3317, which may have beneficial cardiovascular effects.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Androgen receptor suppresses prostate cancer cell invasion via altering the miR-4496/β-catenin signals

Previous study found that AR in prostate may act as both a proliferator and a suppressor to promote or suppress the metastasis of prostate cancer (PCa). In current work, we demonstrated that AR could suppress PCa cell invasion through altering the miR-4496/β-catenin signals. And mechanisms dissection found that AR could negatively regulate the expression of β-catenin through enhancing the miR-4496 expression via directly binding to the AR-response-elements (AREs) of miR-4496 promoter, subsequently, the miRNA could directly target the 3’ UTR of the β-catenin-mRNA to reduce its expression. To conclude, our work suggests that AR might play an important role to suppress PCa cell invasion, targeting the newly identified AR/miR-4496/β-catenin signaling with small molecules may help us to build up new therapeutic approaches to better suppress the metastasis of PCa.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Downregulation of FOXP1 correlates with tendon stem/progenitor cells aging

Aging is known as a major risk factor for tendon disorders whereas the molecular mechanisms of age-related tendon disorders still remains unclear. Since tendon-derived stem/progenitor cells (TSPCs) play a vital role in tendon maintenance and healing, in this study we aimed to investigate the role of Forkhead box P1 (FOXP1) in aged TSPCs, we found that FOXP1 mRNA and protein levels were markedly decreased in the aged TSPCs. Moreover, overexpression of FOXP1 attenuates TSPCs aging, as confirmed by decreased of senescence-associated β-gal staining, as well as the senescence marker, p16{sup INK4A}. Conversely, FOXP1 depletion by siRNA promoted senescence in young TSPCs. Meanwhile, FOXP1 overexpression also restores the age-associated reduction of self-renewal, migration and differentiation of TSPCs. In addition, FOXP1 overexpression rescued decreased levels of E2F1, pRb and cyclin D1 in aged TSPCs, which suggested that FOXP1 regulates TSPCs aging through cellular senescence. These results indicate that FOXP1 plays a crucial role in TSPCs aging.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Norepinephrine-induced downregulation of GLT- 1 mRNA in rat astrocytes

Highlights: • GLT-1 decreased bilaterally in rat spinal cord following unilateral nerve injury. • GLT-1 in cultured astrocytes was downregulated by norepinephrine. • Downregulation of GLT-1 is mediated by the α{sub 1}-adrenoceptor. Glutamate transporter-1 (GLT-1; also known as excitatory amino acid transporter 2) plays an important role in the maintenance of glutamate homeostasis in the synaptic cleft. Downregulation of GLT-1 in the spinal cord has been reported in chronic pain models, which suggests that GLT-1 is involved in the development of chronic pain. However, the mechanism by which GLT-1 is downregulated in the spinal cord is still unknown. We hypothesized that norepinephrine is involved in the regulation of GLT-1. The aim of this study was to investigate the effect of norepinephrine on GLT-1 expression in cultured astrocytes.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

CCN3 secreted by prostaglandin E2 inhibits intimal cushion formation in the rat ductus arteriosus

Highlights: • CCN3 is secreted from smooth muscle cells of the ductus arteriosus by prostaglandin E{sub 2}. • Nano-flow LC-MS/MS analysis is a powerful tool for detecting a novel secreted protein. • CCN3 inhibits intimal cushion formation in the ductus arteriosus. The ductus arteriosus (DA), an essential fetal shunt between the pulmonary trunk and the descending aorta, changes its structure during development. Our previous studies have demonstrated that prostaglandin E{sub 2} (PGE{sub 2})-EP4 signaling promotes intimal cushion formation (ICF) by activating the migration of DA smooth muscle cells via the secretion of hyaluronan. We hypothesized that, in addition to hyaluronan, PGE{sub 2} may secrete other proteins that also regulate vascular remodeling in the DA. In order to detect PGE{sub 2} stimulation-secreted proteins, we found that CCN3 protein was increased in the culture supernatant in the presence of PGE{sub 2} in a dose-dependent manner by nano-flow liquid chromatography coupled with tandem mass spectrometry analysis and enzyme-linked immunosorbent assay. Quantitative RT-PCR analysis revealed that PGE{sub 2} stimulation tended to increase the expression levels of CCN3 mRNA in DA smooth muscle cells. Immunohistochemical analysis revealed that CCN3 was highly localized in the entire smooth muscle layers and the endothelium of the DA. Furthermore, exogenous CCN3 inhibited PGE{sub 2}-induced ICF in the ex vivo DA tissues. These results suggest that CCN3 is a secreted protein of the DA smooth muscle cells induced by PGE{sub 2} to suppress ICF of the DA. The present study indicates that CCN3 could be a novel negative regulator of ICF in the DA to fine-tune the PGE{sub 2}-mediated DA remodeling.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

As an independent prognostic factor, USP6 promotes the invasion and metastasis of colon cancer

Highlights: • USP6 is highly expressed in colon cancer tissues at both mRNA and protein levels. • High USP6 expression was closely associated with the invasion and metastasis of colon cancer. • USP6 overexpression promotes colon cancer cell invasion and metastasis both in vitro and in vivo. • USP6 is considered as a novel gene in promoting the invasion and metastasis of colon cancer. The lack of efficient tumor invasion and metastatic biomarkers led to high mortality rates in colon cancer patients. Aberrant expression of ubiquitin-specific protease 6 (USP6) was involved in several diseases including cancer, while its role in the progression of colon cancer was still unclear. In this study, USP6 was evaluated at both mRNA and protein levels by using RT-PCR, western blot and immunohistochemistry staining analyses. The results revealed that high USP6 expression predicted poor disease-specific survival and overall survival through Kaplan-Meier analyses with log-rank tests, univariate and multivariate Cox analyses. Furthermore, cell function assay demonstrated that USP6 could promote colon cancer cells’ invasion in vitro and liver metastasis in vivo. These findings indicated that high USP6 expression contributed to the progression of colon cancer and USP6 may be a valuable prognostic factor in patients with colon cancer.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

The ADP-ribose reactive NUDIX hydrolase isoforms can modulate HIF-1α in cancer cells

Highlights: • Knockdown of some ADP-ribose reactive NUDIX hydrolases enhances HIF-1α accumulation. • Neither HIF-1 α mRNA expression nor protein degradation is affected by NUDT9 knockdown. • Knockdown of NUDT9 enhances HIF-1α translation via TRPM2-CaMKKβ-AMPK pathway. The human nucleoside-diphosphate linked moiety–X (NUDIX) hydrolases that utilize ADP-ribose and NADH/NAD{sup +} are overexpressed in cancer cells, but their roles in hypoxia inducible factor-1α (HIF-1α) regulation have not yet been revealed. Here, we showed that these NUDIX hydrolases negatively regulated HIF-1α accumulation by modulating the Ca{sup 2+} dependent AMP-activated protein kinase (AMPK) signaling pathway. In specific, knockdown of NUDT9 resulted in accumulation of free ADP-ribose that triggered Ca{sup 2+} influx mediated by transient receptor potential cation channel subfamily M member 2 and subsequent activation of Ca{sup 2+}/calmodulin-dependent protein kinase kinase β (CaMKKβ). In addition, AMPK activation by CaMKKβ was shown to enhance HIF-1α accumulation. Our findings provide insights into the action of NUDIX hydrolases as an additional, discrete modulator of HIF-1α accumulation.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

MicroRNA-98 regulates hepatic cholesterol metabolism via targeting sterol regulatory element-binding protein 2

Highlights: • The expression of miR-98 decreased while the expression of SREBP-2 increased in hypercholesterolemic patients. • SREBP-2 is a direct target gene of miR-98. • The effect of miR-98 on cholesterol metabolism is mediated by SREBP-2. • Overexpression of miR-98 reduced cholesterol level in vitro and in vivo. Hypercholesterolemia is an important risk factor for coronary heart disease. Although a lot of research has been conducted, the regulation of cholesterol metabolism is still largely unknown. Some miRNAs have been found to play critical role in the cholesterol metabolism. MiR-98 is a miRNA whose function has been reported mainly in tumorigenesis. In this study, we elucidate a novel role of miR-98 in cholesterol metabolism. We found that the expression of miR-98 was decreased significantly in hypercholesterolemic patients compared with healthy control subjects. Furthermore, we identified that SREBP-2, an important transcriptional factor in cholesterol metabolism, was a direct target of miR-98. Overexpression of miR-98 significantly repressed the 3′-UTR reporter activities of SREBP-2 in a dose-dependent manner in HepG2 cells, while the effect of miR-98 was blocked when the binding site of miR-98 within the SREBP-2 3′-UTR was mutated. And overexpression of miR-98 reduced both the mRNA and protein levels of HMGCR and LDLR significantly in vitro, which are two target genes of SREBP-2. Furthermore, MiR-98 overexpression reduced the intracellular total cholesterol levels dramatically. Moreover, we overexpressed the miR-98 by lentiviral tail vein injection in vivo. Compared with the control mice, the miR-98 overexpression mice showed lower serum cholesterol level and decreased SREBP-2, HMGCR as well as LDLR expression. Our data confirmed that reduced expression of miR-98 potentially contributes to disturbance of cholesterol metabolism. MiR-98 might be a novel therapeutic target to hypercholesterolemia.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Alleviation of A disintegrin and metalloprotease 10 (ADAM10) on thromboangiitis obliterans involves the HMGB1/RAGE/ NF-κB pathway

Highlights: • TAO rats displayed higher whole blood viscosity and blood platelet count. • TAO rats had structural damages of vascular smooth muscle and endothelial cells. • ADAM treatment alleviated the symptoms of TAO rats in a dose-dependent pattern. • HMGB1, RAGE and NF-κB increased in TAO rats in both mRNA and protein level. • HMGB1, RAGE and NF-κB decreased by ADAM10 treatment in both mRNA and protein level. Thromboangiitis obliterans (TAO), also known as Buerger's disease, is a nonatherosclerotic inflammatory disease that influences medium- and small-sized blood vessels of extremities. However, mechanisms underlying TAO are still unclear. As a mediator associated with inflammation, A disintegrin and metalloprotease 10 (ADAM10) was hypothesized to play inhibitory roles in the development of TAO. Thus, the objective of this study is to investigate the effects of ADAM10 in a sodium laurate-induced TAO rat model and elucidate underlying mechanisms. Male Wistar rats were randomly divided into four groups (n = 6) for treatment: sham-operated (SHAM), TAO model (TAO), ADAM10 low dose injection (3 mg/kg; ADAM10-LD) and ADAM10 high dose injection (6 mg/kg; ADAM10-HD). After 14-day treatment, color Doppler ultrasound and hematology analysis indicated TAO rats displayed higher whole blood viscosity and blood platelet count compared with those in the SHAM group. Histologic evaluation and transmission electron microscopy revealed that the ultrastructural damages of vascular smooth muscle and endothelial cells were observed in TAO rats, such as fractured endoplasmic reticulum, decreased cell counts, and fibrillation. On the other hand, the typical signs and symptoms of TAO rats were significantly alleviated via ADAM10 treatment with a dose-dependent pattern. Real-time PCR and western blot results revealed that the expression of high-mobility-group box 1 (HMGB1), receptor for advanced glycation end-products (RAGE) and nuclear factor-kappa B (NF-κB) increased in TAO rats whereas decreased by ADAM10 treatment in both mRNA and protein levels. In conclusion, the results suggest ADAM10 alleviates symptoms of sodium laurate-induced TAO in rats via the RAGE/NF-κB signaling pathway and provides insight into the molecular basis and a potential therapeutic strategy for TAO.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

ERα is a negative regulator of PD-L1 gene transcription in breast cancer

Highlights: • PD-L1 mRNA level is much lower in ERα-positive breast cancer cell lines. • E2 treatment inhibits PD-L1 gene transcription in ERα-positive MCF7 cells. • Ectopic expression of ERα down-regulates PD-L1 gene expression in ERα-negative MDA-MB-231 cells. • ERα expression is negatively correlated with PD-L1 expression in ERα-positive breast cancer. The programmed death-ligand 1 (PD-L1) expression by tumors results in potent antitumor immune suppression through binding to programmed death-1 (PD-1) on T cells and subsequent inhibition of T cells activity. Although recent pathological studies have shown that PD-L1 is actively expressed in certain ERα-negative breast cancer, little is known about whether ER signaling regulates PD-L1 gene expression. Here, we investigated the relationship between ERα and PD-L1 in breast cancer. Analysis of Comprehensive Cell Line Encyclopedia (CCLE) data showed that the average mRNA level of PD-L1 in ERα-positive breast cancer cell lines was significantly lower than that in ERα-negative breast cancer cell lines. E2 treatment inhibited PD-L1 mRNA expression in hormone-depleted ERα-positive MCF7 cells. Moreover, ectopic expression of ERα in triple-negative MDA-MB-231 cells reduced PD-L1 mRNA and protein expression. Consistently, analysis of The Cancer Genome Atlas (TCGA) data revealed an inverse correlation between ERα and PD-L1 expression in ERα-positive breast cancer. Taken together, our results identify ERα as a negative regulator of PD-L1 gene transcription in breast cancer cells, suggesting that ERα loss-of-function may facilitate the immune evasion of breast cancer cells via up-regulation of PD-L1.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Leonurine hydrochloride promotes osteogenic differentiation and increases osteoblastic bone formation in ovariectomized mice by Wnt/β-catenin pathway

Highlights: • Leonurine hydrochloride promotes osteogenic differentiation of osteoblasts in vitro. • Wnt/β-catenin pathway acts as an important role in the osteogenic differentiation affected by Leonurine hydrochloride. • Leonurine hydrochloride alleviate the effect of osteoporosis by acclerating the bone formation. • Bone mineral density and micro-structures are improved by the treatment of Leonurine hydrochloride. Leonurine hydrochloride (LH) is a synthetic chemical compound derived from leonurine that can be extracted from Leonurus sibiricus and possesses antioxidant, anti-apoptosis, and neuroprotective activities. In previous studies, LH has been demonstrated to attenuate osteoclast activity and prevent bone loss. However, it is unknown whether LH accelerates bone formation and promotes osteogenic differentiation. We systematically examined the effects of LH on ovariectomized-induced osteoporotic mice and the MC3T3-E1 osteoblastic cell line. The results revealed that LH enhanced differentiation of MC3T3-E1 cells, with a dose-dependent increase in alkaline phosphatase (ALP) activity. Moreover, LH upregulated osteogenesis-related gene expression, including osterix, alpha 1 type 1 collagen, runt-related transcription factor 2 (Runx2) and ALP, as shown by quantitative reverse transcription-polymerase chain reaction analysis. At the same time, elevated expression of low-density lipoprotein receptor-related protein 5 and β-catenin mRNA was detected in the Wnt/β-catenin pathway. A western blot analysis revealed that LH dose-dependently increased the expression of Runx2 and β-catenin, and promoted phosphorylation of glycogen synthase kinase-3β in vitro. The in vivo results showed that administering LH (15 mg/kg/d) for 8 weeks alleviated destruction of the trabecular microstructure caused by osteoporosis. LH increased the bone mineral density and trabecular number, decreased trabecular separation according to a micro-computed tomography scan. In addition, LH enhanced the expression of β-catenin and Runx2 in vivo. In conclusion, LH promoted osteogenic differentiation and bone formation in vivo and in vitro, which alleviated osteoporosis through activation of the Wnt/β-catenin pathway.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Indoxyl glucuronide, a protein-bound uremic toxin, inhibits hypoxia-inducible factor‒dependent erythropoietin expression through activation of aryl hydrocarbon receptor

Highlights: • Indoxyl glucuronide (IG) is a protein-bound uremic toxin. • IG inhibited hypoxia-induced HIF transcriptional activation and EPO expression. • IG activated aryl hydrocarbon receptor (AHR) signaling pathway. • The AHR antagonist abolished the inhibitory effect of IG on HIF activation. • IG-induced activation of AHR may contribute to the progression of renal anemia. Renal anemia is common among chronic kidney disease (CKD) patients, and is mainly caused by inadequate erythropoietin (EPO) production from kidneys due to dysfunction of intracellular hypoxia-inducible factor (HIF) signaling in renal EPO-producing cells. We have previously shown that indoxyl sulfate (IS), a representative protein-bound uremic toxin accumulated in the blood of CKD patients, inhibits hypoxia-induced HIF activation and subsequent EPO production through activation of aryl hydrocarbon receptor (AHR). In this study, we further investigated the effects of other protein-bound uremic toxins on HIF-dependent EPO expression using EPO-producing HepG2 cells. We found that indoxyl glucuronide (IG) and IS, but not p-cresyl sulfate, phenyl sulfate, 3-indoleacetic acid or hippuric acid, inhibited hypoxia mimetic cobalt chloride-induced EPO mRNA expression. Furthermore, IG at concentrations similar to the blood levels in CKD patients inhibited the transcriptional activation of HIF induced by both cobalt chloride treatment and hypoxic culture. IG also induced CYP1A1 mRNA expression and nuclear translocation of AHR protein, indicating that IG activates AHR signaling. Blockade of AHR by a pharmacological antagonist CH-223191 abolished the IG-induced inhibition of HIF activation. Collectively, this study is the first to elucidate the biological effects of IG to inhibit HIF-dependent EPO production through activation of AHR. Our data suggests that not only IS but also IG contributes to the impairment of HIF signaling in renal anemia.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Prevention of acute liver injury by suppressing plasma kallikrein-dependent activation of latent TGF-β

Highlights: • TGF-β contributes to the pathogenesis of acute liver injury (ALI). • PLK-dependent TGF-β activation occurs in/around macrophages in ALI mice. • R58 latency associated protein-degradation products serve as an indicator for ALI. • Inhibiting PLK suppresses apoptosis and improves survival rate of ALI mice. Acute liver injury (ALI) is highly lethal acute liver failure caused by different etiologies. Transforming growth factor β (TGF-β) is a multifunctional cytokine and a well-recognized inducer of apoptotic and necrotic cell death in hepatocytes. Latent TGF-β is activated partly through proteolytic cleavage by a serine protease plasma kallikrein (PLK) between the R58 and L59 residues of its propeptide region. Recently, we developed a specific monoclonal antibody to detect the N-terminal side LAP degradation products ending at residue R58 (R58 LAP-DPs) that reflect PLK-dependent TGF-β activation. This study aimed to explore the potential roles of PLK-dependent TGF-β activation in the pathogenesis of ALI. We established a mouse ALI model via the injection of anti-Fas antibodies (Jo2) and observed increases in the TGF-β1 mRNA level, Smad3 phosphorylation, TUNEL-positive apoptotic hepatocytes and R58-positive cells in the liver tissues of Jo2-treated mice. The R58 LAP-DPs were observed in/around F4/80-positive macrophages, while macrophage depletion with clodronate liposomes partly alleviated the Jo2-induced liver injury. Blocking PLK-dependent TGF-β activation using either the serine proteinase inhibitor FOY305 or the selective PLK inhibitor PKSI-527 or blocking the TGF-β receptor-mediated signaling pathway using SB431542 significantly prevented Jo2-induced hepatic apoptosis and mortality. Furthermore, similar phenomena were observed in the mouse model of ALI with the administration of acetaminophen (APAP). In summary, R58 LAP-DPs reflecting PLK-dependent TGF-β activation may serve as a biomarker for ALI, and targeting PLK-dependent TGF-β activation has potential as a therapeutic strategy for ALI.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Hepatic conditional knockout of ATF6 exacerbates liver metabolic damage by repressing autophage through MTOR pathway

Highlights: • Conditional knockout of ATF6 exacerbates liver metabolic damage. • Conditional knockout of ATF6 represses autophage. • Conditional knockout of ATF6 activates MTOR pathway. The liver is a central metabolic organ. Activating transcription factor 6 (ATF6) acts as an ER stress responsive gene and is reported to attenuate hepatic steatosis. Over expressing a dominant-negative form of ATF6 exacerbates glucose intolerance and insulin resistance. In the present study, we used the conditional knockout technique to specifically knockout ATF6 in the mouse liver. We used qPCR to detect the mRNA levels of related genes. Western blot analysis was used to evaluate protein levels. Flow cytometry assay showed the apoptosis status. Glucose tolerance tests and insulin tolerance tests were used to determine glucose and insulin sensitivity. The results showed that liver specific knockout of ATF6 exacerbated HFD-induced hepatic steatosis and glucose tolerance. Abolished ATF6 exacerbated gluconeogenic metabolism by MTOR mediated down regulation of autophage. In conclusion, these findings suggest that therapeutic strategies by supplementing ATF6 may be beneficial for the treatment of glucose intolerance as well as insulin resistance in the high fat induced liver metabolic damage condition.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

MiR-326 antagomir delays the progression of age-related cataract by upregulating FGF1-mediated expression of betaB2-crystallin

Highlights: • FGF-1 upregulates the expression of an anti-cataract protein βB2. • miR-326 inhibits FGF-1 expression and thereby decreases βB2 level. • miR-326 antagomir delay cataract progression via the function of miR-326. Age-related cataract, the most common cause of blindness worldwide, has been found closely associated with β-crystallin B2 (βB2 or CRYBB2). MicroRNAs (miRNAs) are the primary epigenetic regulators important for various biological processes. However, the role of miRNAs in the progression of lens cataract remains to be elucidated. In this study, we found a novel signal cascade miR-326–fibroblast growth factor 1 (FGF1)–βB2 modulating the progression of lens cataract. In brief, miR-326 exacerbated but its antagomirs attenuated H{sub 2}O{sub 2}-induced apoptosis of HLEC-B3 human lens epithelial cells. Dual-luciferase reporter assay and Western blot showed that miR-326 inhibited FGF1 expression by directly targeting its mRNA 3′-UTR. Consistent with this result, miR-326 antagomir enhanced FGF1 protein level. In addition to FGF1, miR-326 antagomir also enhanced βB2 expression and this enhancement was abolished by transfection of HLEC-B3 cells with FGF1 shRNA. These data demonstrated that miR-326 antagomir increased βB2 expression via upregulating FGF1, which was further confirmed by the studies in a rat model of selenite-induced cataract. This work suggests that miR-326 antagomir might be a promising candidate to prevent progression of age-related cataract.

60 APPLIED LIFE SCIENCES↗