Baicalin Inhibits Coxsackievirus B3 Replication by Reducing Cellular Lipid Synthesis

2020 ◽  
Vol 48 (01) ◽  
pp. 143-160 ◽  
Author(s):  
Meng-Jie Wang ◽  
Chun-Hua Yang ◽  
Yue Jin ◽  
Chang-Biao Wan ◽  
Wei-He Qian ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Viral Replication ◽  
Lipid Content ◽  
Hela Cells ◽  
Lipid Synthesis ◽  
Heart Weight ◽  
Mrna Levels ◽  
Cellular Lipid ◽  
Autophagosome Formation ◽  
Cvb3 Infection

Baicalin is a flavonoid extracted from Scutellariae Radix and shows a variety of biological activities as reducing lipids, diminishing inflammation, and inhibiting bacterial infection. However, there is no report of baicalin against CVB3 infection. In this study, we found that baicalin can reduce viral titer in a dose-dependent manner in vitro at a dose with no direct virucidal effect. Moreover, we revealed that baicalin can also improve survival rate, reduce heart weight/body weight ratio, prevent virus replication, and relieve myocardial inflammation in the acute viral myocarditis mouse model induced by CVB3. Then, in order to explore the mechanism of baicalin inhibiting CVB3 replication, we respectively examined the expression of autophagosome marker LC3-II by Western blot, tested the concentration of free fatty acid (FFA) and cholesterol (CHO) by commercial kits, detected the mRNA levels of fatty acid synthase (Fasn) and acetyl coenzyme a carboxylase (ACC) by RT-PCR, and observed the lipid content of cells by fluorescence staining. The results showed that CVB3 infection increased autophagosome formation and lipid content in HeLa cells, but these changes were significantly blocked by baicalin. Finally, in order to confirm that baicalin inhibits viral replication and reduces autophagosome formation by reducing cellular lipids, we added exogenous palmitate to cell culture supernatants to promote intracellular lipid synthesis and found that palmitate did not alter LC3-II and CVB3/VP1 expression in HeLa cells with or without CVB3 infection. Interestingly, palmitate can reverse the inhibitory effect of baicalin on autophagosome formation and viral replication. In conclusion, our results indicated that lipids play an important role in CVB3 replication, and the effect of baicalin against CVB3 was associated with its ability to reduce cellular lipid synthesis to limit autophagosome formation.

Tanshinone IIA Promotes Macrophage Cholesterol Efflux and Attenuates Atherosclerosis of apoE-/- Mice by Omentin-1/ABCA1 Pathway

2019 ◽  
Vol 20 (5) ◽  
pp. 422-432 ◽  
Author(s):  
Yu-lin Tan ◽  
Han-xiao Ou ◽  
Min Zhang ◽  
Duo Gong ◽  
Zhen-wang Zhao ◽  
...  
Keyword(s):  
Atherosclerotic Plaque ◽  
Lipid Content ◽  
Plasma Levels ◽  
Cholesterol Efflux ◽  
Density Lipoprotein ◽  
Tanshinone Iia ◽  
Lipoprotein Cholesterol ◽  
Mrna Levels ◽  
Cellular Lipid ◽  
Oil Red O

Background: Tanshinone IIA (Tan IIA) and Omentin-1 have a protective role in the cardiovascular system. However, if and how Tan IIA and Omentin-1 regulate cholesterol metabolism in macrophages has not been fully elucidated. Objective: To investigate the possible mechanisms of Tan IIA and Omentin-1 on preventing macrophage cholesterol accumulation and atherosclerosis development. Methods: The effect of Tan IIA on the protein and mRNA levels of Omentin-1 and ATP-binding cassette transporter A1 (ABCA1) in macrophages was examined by Western blot and qRT-PCR assay, respectively. Cholesterol efflux was assessed by liquid scintillation counting (LSC). Cellular lipid droplet was measured by Oil Red O staining, and intracellular lipid content was detected by high performance liquid chromatography (HPLC). In addition, the serum lipid profile of apoE−/− mice was measured by enzymatic method. The size of atherosclerotic lesion areas and content of lipids and collagen in the aortic of apoE−/− mice were examined by Sudan IV, Oil-red O, and Masson staining, respectively. Results: Tan IIA up-regulated expression of Omentin-1 and ABCA1 in THP-1 macrophages, promoting ABCA1-mediated cholesterol efflux and consequently decreasing cellular lipid content. Consistently, Tan IIA increased reverse cholesterol transport in apoE−/− mice. Plasma levels of high-density lipoprotein cholesterol (HDL-C), ABCA1 expression and atherosclerotic plaque collagen content were increased while plasma levels of low-density lipoprotein cholesterol (LDL-C) and atherosclerotic plaque sizes were reduced in Tan IIA-treated apoE−/− mice. These beneficial effects were, however, essentially blocked by knockdown of Omentin-1. Conclusion: Our results revealed that Tan IIA promotes cholesterol efflux and ameliorates lipid accumulation in macrophages most likely via the Omentin-1/ABCA1 pathway, reducing the development of aortic atherosclerosis.

scholarly journals miR-21-5p regulates mitochondrial respiration and lipid content in H9C2 cells

2019 ◽  
Vol 316 (3) ◽  
pp. H710-H721 ◽  
Author(s):  
Victoria L. Nasci ◽  
Sandra Chuppa ◽  
Lindsey Griswold ◽  
Kathryn A. Goodreau ◽  
Ranjan K. Dash ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Fatty Acid ◽  
Palmitic Acid ◽  
Fatty Acid Oxidation ◽  
Lipid Content ◽  
Mitochondrial Respiration ◽  
Acid Oxidation ◽  
H9c2 Cells ◽  
Cellular Lipid ◽  
Transfected Cells

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty acid oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate lipid content, lipid peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before lipid treatment in culture or as part of the Agilent Seahorse XF fatty acid oxidation assay. Overexpression of miR-21-5p attenuated the lipid-induced increase in cellular lipid content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of lipid peroxidation, was significantly increased with lipid treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and palmitic acid. Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and palmitic acid, and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both lipid content and lipid peroxidation in H9C2 cells. This likely occurs by reducing cellular lipid uptake and utilization, shifting cellular metabolism toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty acid oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

scholarly journals Lipid Dynamics, Identification, and Expression Patterns of Fatty Acid Synthase Genes in an Endoparasitoid, Meteorus pulchricornis (Hymenoptera: Braconidae)

2020 ◽  
Vol 21 (17) ◽  
pp. 6228
Author(s):  
Jiao Wang ◽  
Li-Wei Shen ◽  
Xiao-Rong Xing ◽  
Yu-Qi Xie ◽  
Yi-Jiangcheng Li ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipid Content ◽  
Fatty Acid Synthase ◽  
Developmental Stages ◽  
Adult Stage ◽  
Expression Patterns ◽  
Lipid Synthesis ◽  
Open Reading Frames ◽  
Lipid Dynamics ◽  
Meteorus Pulchricornis

In insect parasitoids, fatty acid synthases (FASs) have received less attention and their roles associated with lipogenesis loss are far from clear. Meteorus pulchricornis is a solitary endoparasitoid wasp of many larvae of lepidopteran pests. The lipid content during developmental stages of M. pulchricornis was measured; it was higher in the larval and pupal stages but declined from six-day-old pupae. Lipid accumulation constantly decreased in the adult stage, even after feeding on honey solutions. To investigate the roles of FASs in lipid synthesis in M. pulchricornis, four FAS genes (MpulFAS1~4) were identified from the transcriptome database of M. pulchricornis. All FAS genes included full-length open reading frames and shared 72–79% similarity with the sequences of Microplitis demolitor. qRT-PCR validation showed that all four FASs had the highest expression after the adult wasps were fed on honey diets. MpulFAS1 and MpulFAS2 reached their expression peaks at the adult stage but MpulFAS3 and MpulFAS4 peaked at the larval stage. To further study the function of FASs, dsRNA injection knocked down the expression of four MpulFASs and resulted in a significant decline of lipid content at the adult stage in M. pulchricornis. Results from this study suggest that M. pulchricornis adults cannot accumulate lipid content effectively and FASs may still contribute to lipid synthesis in the adult stage. This broadens the knowledge on the ability of lipid synthesis in parasitoid wasps and provides insight into the roles of FASs in insects with parasitic life-history traits.

P–208 Oleic acid rescues altered autophagy induced by palmitic acid during mouse preimplantation development

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Z Leung ◽  
M Calder ◽  
D Betts ◽  
B Ab. Rafea ◽  
A Watson
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Preimplantation Development ◽  
Preimplantation Embryos ◽  
Mrna Levels ◽  
Autophagosome Formation ◽  
Fatty Acid Treatment ◽  
Autophagic Activity ◽  
Lc3 Puncta

Abstract Study question The aim of the study is to identify the autophagic profile and the effects of fatty acid treatments on autophagic activity in preimplantation mouse embryos. Summary answer Autophagic activity varies significantly in early stages of mouse preimplantation development; exposure to fatty acids alters the embryonic autophagy profile. What is known already Obesity is one of the top comorbidities for infertility, and obese individuals have elevated fatty acid levels. In serum, palmitic acid (PA) and oleic acid (OA) are the most abundant saturated and unsaturated fatty acids, respectively. We recently reported that PA impairs blastocyst development, affects mitochondrial reactive oxygen species, triacylglycerol levels, and endoplasmic reticulum stress pathways during mouse preimplantation development. Interestingly, the addition of OA counteracts those effects. Autophagy plays an essential role in embryo development, as knock-out of a key autophagy protein is embryonic lethal. Little is known about the autophagic profile in fatty acid treated mouse preimplantation embryos. Study design, size, duration Pools of 20 – 25 mouse embryos were collected from gonadotrophin super-ovulated and mated CD1 female mice. Two-cell stage embryos were treated with 100 µM PA and 250 µM OA, alone and in combination, and 1.5% bovine serum albumin media (control) within KSOMaa media for 18, 24, and 48 hours in vitro. The detection of various autophagic markers were evaluated by immunofluorescence microscopy and RT-qPCR. Participants/materials, setting, methods mRNA levels of autophagic markers were measured using RT-qPCR with the Taqman primers and Universal PCR Mix. Immunofluorescence staining of LC3 puncta (marker for autophagosome formation) was performed using LC3A/B polyclonal antibody (Invitrogen PA1–16931) and DAPI (4′,6-Diamidino–2-phenylindole dihydrochloride) was used to stain for cell nuclei. Analysis of LC3 puncta was performed using ImageJ software. Images were acquired using an LSM 800 laser scanning confocal microscope. Data analysis was completed by GraphPad Prism software. Main results and the role of chance Mouse preimplantation embryos showed no change in mRNA levels of autophagic markers (Bcln1, ATG3, ATG5, and LC3) relative to the control group after 48-hours exposure of 100 µM PA and 250 µM OA treatments, alone and in combination. The number of LC3 puncta was measured and analyzed as a reflection of autophagic activity in mouse preimplantation embryos. Under the fatty acid-free condition, the average number of LC3 puncta per blastomere was significantly decreased after 18 hours of development (p < 0.005). However, the average number of LC3 puncta per blastomere at 18, 24, and 48 hours were not significantly different from each other (p = 0.2724). Following 100 µM PA and 250 µM OA treatments, alone and in combination, autophagic activity was impacted by the presence of fatty acids. Mouse preimplantation embryos exposed to control and fatty acid treatment groups demonstrated no significant differences in LC3 puncta per blastomere at 18- and 24-hours treatment time (p = 0.5381; p = 0.7829). However, embryos exposed to 48 hours of PA treatment had a significantly greater number of LC3 puncta per blastomere than embryos exposed to 48 hours of OA and PA and OA combination treatments (p < 0.05). Limitations, reasons for caution Although LC3 puncta count (autophagosome formation) is impacted by fatty acid treatment, autophagic flux must be measured to fully investigate autophagic activity during mouse preimplantation development. These processes need to be measured in human embryos cultured in vitro. Wider implications of the findings: Profiling autophagic activity in fatty acid treated mouse preimplantation embryos would guide future investigations on pharmacological modulation of autophagy as a therapeutic intervention for developmentally delayed embryos. With the information gained, we aim to develop strategies to assist overweight and obese patients with their fertility needs. Trial registration number Not applicable

scholarly journals 25-Hydroxycholesterol-3-sulfate regulates macrophage lipid metabolism via the LXR/SREBP-1 signaling pathway

2008 ◽  
Vol 295 (6) ◽  
pp. E1369-E1379 ◽  
Author(s):  
Yongjie Ma ◽  
Leyuan Xu ◽  
Daniel Rodriguez-Agudo ◽  
Xiaobo Li ◽  
Douglas M. Heuman ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipid Metabolism ◽  
Fatty Acid Synthase ◽  
Lipid Synthesis ◽  
Mrna Levels ◽  
Intracellular Lipids ◽  
Protein Levels ◽  
Regulatory Molecule ◽  
Fas Mrna

The oxysterol receptor LXR is a key transcriptional regulator of lipid metabolism. LXR increases expression of SREBP-1, which in turn regulates at least 32 genes involved in lipid synthesis and transport. We recently identified 25-hydroxycholesterol-3-sulfate (25HC3S) as an important regulatory molecule in the liver. We have now studied the effects of 25HC3S and its precursor, 25-hydroxycholesterol (25HC), on lipid metabolism as mediated by the LXR/SREBP-1 signaling in macrophages. Addition of 25HC3S to human THP-1-derived macrophages markedly decreased nuclear LXR protein levels. 25HC3S administration was followed by dose- and time-dependent decreases in SREBP-1 mature protein and mRNA levels. 25HC3S decreased the expression of SREBP-1-responsive genes, acetyl-CoA carboxylase-1, and fatty acid synthase (FAS) as well as HMGR and LDLR, which are key proteins involved in lipid metabolism. Subsequently, 25HC3S decreased intracellular lipids and increased cell proliferation. In contrast to 25HC3S, 25HC acted as an LXR ligand, increasing ABCA1, ABCG1, SREBP-1, and FAS mRNA levels. In the presence of 25HC3S, 25HC, and LXR agonist T0901317, stimulation of LXR targeting gene expression was repressed. We conclude that 25HC3S acts in macrophages as a cholesterol satiety signal, downregulating cholesterol and fatty acid synthetic pathways via inhibition of LXR/SREBP signaling. A possible role of oxysterol sulfation is proposed.

scholarly journals Depot-Specific Modulation of Rat Intraabdominal Adipose Tissue Lipid Metabolism by Pharmacological Inhibition of 11β-Hydroxysteroid Dehydrogenase Type 1

Endocrinology ◽  
2007 ◽  
Vol 148 (5) ◽  
pp. 2391-2397 ◽  
Author(s):  
Magalie Berthiaume ◽  
Mathieu Laplante ◽  
William Festuccia ◽  
Yves Gélinas ◽  
Sébastien Poulin ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Fatty Acid ◽  
Lipid Metabolism ◽  
Food Intake ◽  
Lipid Synthesis ◽  
Hydroxysteroid Dehydrogenase ◽  
Mrna Levels ◽  
Compound A ◽  
Carnitine Palmitoyltransferase 1

The metabolic consequences of visceral obesity have been associated with amplification of glucocorticoid action by 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in adipose tissue. This study aimed to assess in a rat model of diet-induced obesity the effects of pharmacological 11β-HSD1 inhibition on the morphology and expression of key genes of lipid metabolism in intraabdominal adipose depots. Rats fed a high-sucrose, high-fat diet were treated or not with a specific 11β-HSD1 inhibitor (compound A, 3 mg/kg·d) for 3 wk. Compound A did not alter food intake or body weight gain but specifically reduced mesenteric adipose weight (−18%) and adipocyte size, without significantly affecting those of epididymal or retroperitoneal depots. In mesenteric fat, the inhibitor decreased (to 25–50% of control) mRNA levels of genes involved in lipid synthesis (FAS, SCD1, DGAT1) and fatty acid cycling (lipolysis/reesterification, ATGL and PEPCK) and increased (30%) the activity of the fatty acid oxidation-promoting enzyme carnitine palmitoyltransferase 1. In striking contrast, in the epididymal depot, 11β-HSD1 inhibition increased (1.5–5-fold) mRNA levels of those genes related to lipid synthesis/cycling and slightly decreased carnitine palmitoyltransferase 1 activity, whereas gene expression remained unaffected in the retroperitoneal depot. Compound A robustly reduced liver triacylglycerol content and plasma lipids. The study demonstrates that pharmacological inhibition of 11β-HSD1, at a dose that does not alter food intake, reduces fat accretion specifically in the mesenterical adipose depot, exerts divergent intraabdominal depot-specific effects on genes of lipid metabolism, and reduces steatosis and lipemia.

scholarly journals Autophagosome Supports Coxsackievirus B3 Replication in Host Cells

2008 ◽  
Vol 82 (18) ◽  
pp. 9143-9153 ◽  
Author(s):  
Jerry Wong ◽  
Jingchun Zhang ◽  
Xiaoning Si ◽  
Guang Gao ◽  
Ivy Mao ◽  
...  
Keyword(s):  
Viral Replication ◽  
Protein Degradation ◽  
Viral Myocarditis ◽  
Membrane Vesicles ◽  
Coxsackievirus B3 ◽  
Host Cells ◽  
Small Interfering Rnas ◽  
Autophagosome Formation ◽  
Cvb3 Infection

ABSTRACT Recent studies suggest a possible takeover of host antimicrobial autophagy machinery by positive-stranded RNA viruses to facilitate their own replication. In the present study, we investigated the role of autophagy in coxsackievirus replication. Coxsackievirus B3 (CVB3), a picornavirus associated with viral myocarditis, causes pronounced intracellular membrane reorganization after infection. We demonstrate that CVB3 infection induces an increased number of double-membrane vesicles, accompanied by an increase of the LC3-II/LC3-I ratio and an accumulation of punctate GFP-LC3-expressing cells, two hallmarks of cellular autophagosome formation. However, protein expression analysis of p62, a marker for autophagy-mediated protein degradation, showed no apparent changes after CVB3 infection. These results suggest that CVB3 infection triggers autophagosome formation without promoting protein degradation by the lysosome. We further examined the role of the autophagosome in CVB3 replication. We demonstrated that inhibition of autophagosome formation by 3-methyladenine or small interfering RNAs targeting the genes critical for autophagosome formation (ATG7, Beclin-1, and VPS34 genes) significantly reduced viral replication. Conversely, induction of autophagy by rapamycin or nutrient deprivation resulted in increased viral replication. Finally, we examined the role of autophagosome-lysosome fusion in viral replication. We showed that blockage of the fusion by gene silencing of the lysosomal protein LAMP2 significantly promoted viral replication. Taken together, our results suggest that the host's autophagy machinery is activated during CVB3 infection to enhance the efficiency of viral replication.

scholarly journals Liver X Receptors Suppress Activity of Cholesterol and Fatty Acid Synthesis Pathways To Oppose Gammaherpesvirus Replication

mBio ◽  
2018 ◽  
Vol 9 (4) ◽  
Author(s):  
P. T. Lange ◽  
C. Schorl ◽  
D. Sahoo ◽  
V. L. Tarakanova
Keyword(s):  
Fatty Acid ◽  
Viral Replication ◽  
Metabolic Pathways ◽  
Cholesterol Synthesis ◽  
Target Genes ◽  
Adult Population ◽  
Lipid Synthesis ◽  
Type I ◽  
Liver X Receptors ◽  
X Receptors

ABSTRACTGammaherpesviruses are oncogenic pathogens that persist in ~95% of the adult population. Cellular metabolic pathways have emerged as important regulators of many viral infections, including infections by gammaherpesviruses that require several lipid synthetic pathways for optimal replication. Liver X receptors (LXRs) are transcription factors that are critical regulators of cellular fatty acid and cholesterol synthesis pathways. Not surprisingly, LXRs are attractive therapeutic targets in cardiovascular disease. Here we describe an antiviral role for LXRs in the context of gammaherpesvirus infection of primary macrophages. We show that type I interferon increased LXR expression following infection. Surprisingly, there was not a corresponding induction of LXR target genes. Rather, LXRs suppressed the expression of target genes, leading to decreased fatty acid and cholesterol synthesis, two metabolic pathways that support gammaherpesvirus replication. This report defines LXR-mediated restriction of cholesterol and lipid synthesis as an intrinsic metabolic mechanism to restrict viral replication in innate immune cells.IMPORTANCEFatty acid and cholesterol synthesis pathways of the host play important roles in diverse biological systems. Importantly, these two metabolic pathways are also usurped by a number of viruses to facilitate viral replication. In this report, we show that suppression of these pathways by liver X receptors in primary macrophages creates an intrinsic antiviral state that attenuates gammaherpesvirus replication by limiting viral access to the two metabolic pathways.

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