Spectrofluorometric studies of the lipid probe, nile red.

Greenspan, Phillip; Fowler, S

doi:10.1016/s0022-2275(20)34307-8

Cited by 840 publications

(396 citation statements)

References 23 publications

Supporting

Mentioning

352

Contrasting

Order By: Relevance

“…Thus, to explore the role of HBA in lipid-lowering and anti-oxidation in different disease progressions of NAFLD, larval zebrafish were induced by HCD for 7 or 14 days in vivo. The level of whole-body lipid in larval zebrafish was measured by Nile red staining, which is a fluorochrome staining neutral lipid (Greenspan and Fowler, 1985). Lipid accumulation in the liver was demonstrated by HE staining.…”

Section: Discussionmentioning

confidence: 99%

“…To explore the role of HBA in isolated steatosis and mild nonalcoholic steatohepatitis, larval zebrafish models were studied at 7 and 14 days, respectively, by the inducement of HCD, as in a previous study (Ji et al, 2019) (Figure 2A). The effect of HAB in regulating lipid metabolism was measured by Nile Red staining, which is a red phenoxazine lipid fluorescent dye that can label the neutral lipid properties (Greenspan and Fowler, 1985). As the Nile Red results have shown, the intensity of Nile Red fluorescence within the model group increased gradually in the whole trunk and liver over time by the inducement of HCD compared with the Control.…”

Section: Effect Of P-hydroxybenzyl Alcohol In Regulating Lipid Metabolism On High-cholesterol Diet-induced Larval Zebrafish Modelmentioning

confidence: 99%

See 1 more Smart Citation

P- Hydroxybenzyl Alcohol Alleviates Oxidative Stress in a Nonalcoholic Fatty Liver Disease Larval Zebrafish Model and a BRL-3A Hepatocyte Via the Nrf2 Pathway

Cheng

Yang

et al. 2021

Front. Pharmacol.

View full text Add to dashboard Cite

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease, and it has gradually become the main disease burden in the world. However, the pathogenesis of NAFLD is complex, involving such things as dyslipidemia, oxidative stress, inflammation, etc. This brings to the table a significant challenge for drug development, and there is still no drug approved by the FDA on the market to treat the disease. GAS and HBA are active ingredients of the orchidaceae plant gastrodia elata and have exhibit effects in ameliorating nervous system diseases caused by oxidative stress. HBA is a metabolite of GAS that could perform lipid regulation and improve oxidative stress on HCD-induced NAFLD larval zebrafish, as reported by previous studies; we therefore explored the role of HBA in lipid regulation and oxidative stress on HCD-induced NAFLD larval zebrafish in vivo and FFA-induced BRL-3A hepatocyte in vitro. The gene expression of lipogenesis, inflammation, and oxidative stress were measured to investigate the underlying mechanism of HBA, and the potential protein target of HBA was explored by immunofluorescence. Altogether, our data highlight the role of HBA in improving NAFLD by use of its lipid-lowering and anti-oxidative properties via the Nrf2/HO-1 signaling pathway, providing a potential therapeutic compound for NAFLD treatment.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Effect Of P-hydroxybenzyl Alcohol In Regulating Lipid Metabolism On High-cholesterol Diet-induced Larval Zebrafish Modelmentioning

confidence: 99%

P- Hydroxybenzyl Alcohol Alleviates Oxidative Stress in a Nonalcoholic Fatty Liver Disease Larval Zebrafish Model and a BRL-3A Hepatocyte Via the Nrf2 Pathway

Cheng

Yang

et al. 2021

Front. Pharmacol.

View full text Add to dashboard Cite

show abstract

“…Even though is a well-established and studied model efflux substrate, to counter the possibility that EtBr would give abnormal results which are not representative of other efflux substrates, the same accumulation pattern in Salmonella WT and Δ tolC was also shown using the lipophilic dye Nile Red ( Figure S6 ). Unlike EtBr, Nile Red does not fluoresce on intercalation with DNA, but rather when bound to phospholipids or triglycerides 31 showing this is not an artefact of the dye initially used. Together this data shows that the accumulation pattern described in the absence of efflux is consistent regardless of Gram-negative species, media type or efflux substrate used.…”

Section: Resultsmentioning

confidence: 99%

“…In these experiments SYTO 9 (10 μ M; Thermo Fisher Scientific) was used to differentiate cells from acellular particles using the BL2-H channel. Nile red has an excitation of 549 nm and emission of 628 nm in the presence of phospholipids, and in a neutral lipid environment (tryglycerides), the fluorescence shifts to ex/em of 510/580 nm 31 . Nile red fluorescence was excited using the yellow laser and detected using the YL1-H channel for orange fluorescence 17 .…”

Section: Flow Cytometry Assaymentioning

confidence: 99%

Efflux only impacts drug accumulation in actively growing cells

Trampari

et al. 2021

Preprint

View full text Add to dashboard Cite

For antibiotics with intracellular targets, effective treatment of bacterial infections requires the drug to accumulate to a high concentration inside cells. Bacteria produce a complex cell envelope and possess drug-export efflux pumps to limit drug accumulation inside cells. Decreasing cell envelope permeability and increasing efflux pump activity can reduce intracellular accumulation of antibiotics, and are commonly seen in antibiotic resistant strains. Here, we show that the balance between influx and efflux differs depending on bacterial growth phase in Gramnegative bacteria. Accumulation of the model fluorescent drug, ethidium bromide (EtBr) was measured in S. Typhimurium SL1344 (wild-type) and efflux deficient (ΔacrB) strains during growth. In SL1344, EtBr accumulation remained low, regardless of growth phase and did not correlate with acrAB transcription. EtBr accumulation in ΔacrB was high in exponential phase but dropped sharply later in growth, with no significant difference to SL1344 in stationary phase. Low EtBr accumulation in stationary phase was not due to the upregulation of other efflux pumps, but instead, due to decreased permeability of the envelope in stationary phase. RNAseq identified changes in expression of several pathways that remodel the envelope in stationary phase, leading to lower permeability. This study shows that efflux is only important for maintaining low drug accumulation in actively growing cells, and that envelope permeability is the predominant factor dictating the rate of drug entry in stationary phase cells. This conclusion means that (i) antibiotics with intracellular targets may be less effective in complex non-growing or slow-growing bacterial infections where intracellular accumulation may be low, (ii) efflux inhibitors may be successful in potentiating the activity of existing antibiotics, but potentially only for bacterial infections where cells are actively growing and (iii) the remodelling of the cell envelope prior to stationary phase could provide novel drug targets.

show abstract

“…Another important effect is the sensitivity of the emission spectra of Nile red to the chemical and physical properties of the solvents (Lakowicz, 1983), Nile red concentration (Greenspan and Fowler, 1985), and yeast species (Sitepu et al, 2019), thus excitation and emission wavelengths should also be carefully evaluated to accurately quantify neutral lipids for biotechnological purposes. The wavelengths used in the present study were previously adjusted to allow comparison of the effect of the tested solvents in Nile red.…”

Section: Discussionmentioning

confidence: 99%

Nile Red Incubation Time Before Reading Fluorescence Greatly Influences the Yeast Neutral Lipids Quantification

et al. 2021

View full text Add to dashboard Cite

High-throughput screening methodologies to estimate lipid content in oleaginous yeasts use Nile red fluorescence in a given solvent and optimized excitation/emission wavelengths. However, Nile red fluorescence stabilization has been poorly analyzed, and high variability occurs when relative fluorescence is measured immediately or a few minutes after dye addition. The aim of this work was to analyze the fluorescence of Nile red at different incubation times using a variety of solvents and oleaginous/non-oleaginous yeast strains. We showed that fluorescence stabilization occurs between 20 and 30 min, depending on the strain and solvent. Therefore, we suggest that fluorescence measurements should be followed until stabilization, where Relative Fluorescence Units should be considered after stabilization for lipid content estimation.

show abstract

Spectrofluorometric studies of the lipid probe, nile red.

Cited by 840 publications

References 23 publications

P- Hydroxybenzyl Alcohol Alleviates Oxidative Stress in a Nonalcoholic Fatty Liver Disease Larval Zebrafish Model and a BRL-3A Hepatocyte Via the Nrf2 Pathway

P- Hydroxybenzyl Alcohol Alleviates Oxidative Stress in a Nonalcoholic Fatty Liver Disease Larval Zebrafish Model and a BRL-3A Hepatocyte Via the Nrf2 Pathway

Efflux only impacts drug accumulation in actively growing cells

Nile Red Incubation Time Before Reading Fluorescence Greatly Influences the Yeast Neutral Lipids Quantification

Contact Info

Product

Resources

About