The properties of microparticles from RAW 264.7 macrophage cells undergoing in vitro activation or apoptosis

Recent studies continue to demonstrate the potential of edible insects as a protein base to obtain bioactive peptides applicable for functional food development. This study aimed at identifying antihypertensive, anti-glycemic, and anti-inflammatory peptides derived from the in vitro gastrointestinal digests of cricket protein hydrolysates. After sequential fractionation, the protein digest subfraction containing the lowest molecular weight (<0.5 kDa), hydrophobic (C18) and cationic peptides (IEX) was found responsible for the most bioactivity. The cationic peptide fraction significantly reduced (p < 0.05) α-amylase, α-glucosidase, and angiotensin converting enzyme (ACE) activity in vitro, and also inhibited the expression of NF-κB in RAW 264.7 macrophage cells. A total of 28 peptides were identified with mass spectrometry (LC–MS/MS) and de novo sequencing from the potent fraction. Three novel peptides YKPRP, PHGAP, and VGPPQ were chosen for the molecular docking studies. PHGAP and VGPPQ exhibited a higher degree of non-covalent interactions with the enzyme active site residues and binding energies comparable to captopril. Results from this study demonstrate the bioactive potential of edible cricket peptides, especially as ACE inhibitors.

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Thymol and Piperine-Loaded Poly(D,L-lactic-co-glycolic acid) Nanoparticles Modulate Inflammatory Mediators and Apoptosis in Murine Macrophages

Planta Medica International Open ◽

10.1055/a-1543-1190 ◽

2021 ◽

Vol 8 (03) ◽

pp. e122-e130

Author(s):

Jean Paul Dzoyem ◽

Nathalie Boulens ◽

Eric Allémann ◽

Florence Delie

Keyword(s):

Inflammatory Mediators ◽

Glycolic Acid ◽

In Vitro Assays ◽

Raw 264.7 ◽

Griess Reagent ◽

Macrophage Cells ◽

Raw 264.7 Macrophage ◽

Raw 264.7 Macrophage Cells ◽

Free Molecules

AbstractThis study aimed at preparing and characterizing thymol, eugenol, and piperine-loaded poly(D,L-lactic-co-glycolic acid) nanoparticles and evaluating the effect on inflammatory mediators secretion and apoptosis in Raw 264.7 macrophage cells. Nanoparticles were produced by the solvent evaporation technique. Dynamic light scattering and scanning electron microscopy were used to study the physicochemical characteristics. Raw 264.7 macrophage cells were used as a model for in vitro assays. The 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium assay was used to determine the cytotoxicity of the formulated nanoparticles. An annexin V apoptosis detection kit was used to assess apoptosis. Nitric oxide production was determined using the Griess reagent, and the inflammatory mediators level was evaluated with Th1/Th2 cytokine and fluorometric cyclooxygenase kits. The loaded nanoparticles showed a particle size around 190 nm with a low polydispersity between 0.069 and 0.104 and a zeta potential between–1.2 and–9.5 mV. Reduced cytotoxicity of nanoparticles compared to free molecules against Raw 264.7 macrophage cells was observed and seemed to occur through a mechanism associated with apoptosis. A decrease in cyclooxygenase enzyme activity with an increasing concentration was observed. Both free molecules and nanoparticles showed their capacity to modulate the inflammatory process mostly by inhibiting the investigated inflammatory cytokines. The data presented in this study indicate that thymol and piperine-loaded poly(D,L-lactic-co-glycolic acid nanoparticles could serve as a novel anti-inflammatory colloidal drug delivery system with reduced toxicity. However, further study should be considered to optimize the formulation’s loading capacity and thereby probably enhance their bioactivity in treating inflammatory diseases.

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Açaí (Euterpe oleracea Mart.) reduces the inflammatory response triggered in vitro by the antipsychotic drug olanzapine in RAW 264.7 macrophage cells

Acta Scientiarum Polonorum Technologia Alimentaria ◽

10.17306/j.afs.0857 ◽

2021 ◽

Vol 20 (2) ◽

pp. 149-163

Author(s):

Marcelo S. Fernandes ◽

◽

Alencar K. Machado ◽

Charles E. Assmann ◽

Eduardo N. Andrade ◽

...

Keyword(s):

Inflammatory Response ◽

Antipsychotic Drug ◽

Raw 264.7 ◽

Euterpe Oleracea ◽

Macrophage Cells ◽

Raw 264.7 Macrophage ◽

Raw 264.7 Macrophage Cells ◽

Euterpe Oleracea Mart

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Optimization of an anti-inflammatory screening model on the RAW 264.7 macrophage cell

Science and Technology Development Journal - Natural Sciences ◽

10.32508/stdjns.v1it5.531 ◽

2018 ◽

Vol 1 (T5) ◽

pp. 18-26

Author(s):

To Dinh Le ◽

Kien Trung Nguyen ◽

Thuoc Linh Tran ◽

Thao Thi Phuong Dang

Keyword(s):

New Drugs ◽

No Production ◽

Raw 264.7 ◽

Macrophage Cells ◽

Raw 264.7 Macrophage ◽

Model Cell ◽

Raw 264.7 Macrophage Cells ◽

Anti Inflammatory ◽

Living Tissues

Inflammation is the response of living tissues to the injury. Prolonged or inappropriate inflammation has been involved with many diseases such as, cancer, diabetes, heart disease… These days, inflammation has been treated by nonsteroidal and steroidal anti-inflammatory drugs, which have a lot of side effects. It opens the need and interest of new drugs discovery. Particularly, scientific and pharmaceutical communities show great interest in finding new anti-inflammatory compounds in traditional medicinal plants. This study aimed to optimize an in vitro anti-inflammatory model. Cell heterogeneity, cell density, LPS concentration and LPS incubation time were chosen to optimize the production of nitric oxide (NO) in RAW 264.7 macrophage cells. Our results show that 104 cells/well, FBS 1 %, starvation for 6 h, LPS 0.5 µg/mL in 24 h were optimized parameters in the model. Then, extracts from Hedyotis capitellata, a traditional medicinal plant used by K’ho minority, Bidoup Nuiba national park, Lam Dong province, Vietnam, was chosen to evaluate the in vitro antiinflammatory model. The anti-inflammatory activity was tested by measuring the production of NO in lipopolysaccharide-activated RAW 264.7 macrophage cells. The experimental data indicate that the extracts of this plant decreased NO production in LPS-stimulated RAW 264.7, particularly, the petroleum ether fraction at the concentration of 23.8 µg/mL inhibited NO production by 128.20 %; whereas dexamethasone 50 µM inhibited NO accumulation to 112 %.

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