The Antioxidative Effects of Borago Officinalis in Lipopolysaccharide and Hydrogen Peroxide-Activated RAW 264.7 Macrophages

Abstract Objectives Borage (Borago officinalis) is a plant herb found widely in Asian and Mediterranean regions which has been used for the treatment of chronic conditions and diseases. Gamma-linolenic acid (GLA) and phenolic acids are important constituents of borage, which are known for their antioxidative properties. However, little is known regarding the mechanisms in which borage elicits its antioxidant effects. Therefore, it is important to further examine the antioxidative properties of Borago officinalis extract (BOE) on levels of biomarkers of oxidative stress in lipopolysaccharide (LPS) and hydrogen peroxide (H2O2) stimulated RAW 264.7 macrophages. Methods High-performance liquid chromatography (HPLC) was used to determine the total polyphenolic content of BOE. RAW264.7 murine macrophages were incubated with BOE (0, 50, 100, 200 and 300 µg/ml) followed by treatments with LPS (50 ng/ml) or H2O2 (50 ng/ml) for 24 hours. Media was collected for assessment of nitric oxide (NO), and the cell lysates were collected for determining levels of catalase. BOE treated cells induced with LPS and H2O2 were further examined to assess levels of reactive oxygen species (ROS). Results Cells treated with LPS, H2O2, as well as BOE did not show any decreases in cell viability. The total polyphenolic content of BOE was 102.4 mg/g, with rosmarinic acid the most abundant polyphenol. BOE decreased (P < 0.05) levels of NO when induced with LPS at 300 µg/ml and at dosages of 100, 200, and 300 µg/ml when cells were stimulated with H2O2. The level of catalase was increased (P < 0.05) in H2O2-stimulated macrophages treated with 300 µg/ml BOE. Conclusions This is the first study to our knowledge to mechanistically examine the antioxidative properties of crude BOE in H2O2 and LPS stimulated RAW 264.7 macrophages. These findings indicate that BOE is efficacious as an antioxidative agent which can be used as an alternative or adjuvant therapy. Further research is needed to determine the benefits of BOE's polyphenolic profile and GLA to isolate constituents of interest. Funding Sources None.

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Taurine chloramine protects RAW 264.7 macrophages against hydrogen peroxide-induced apoptosis by increasing antioxidants

Journal of Clinical Biochemistry and Nutrition ◽

10.3164/jcbn.10-120 ◽

2011 ◽

Vol 49 (1) ◽

pp. 50-56 ◽

Cited By ~ 18

Author(s):

Shuyu Piao ◽

Young-Nam Cha ◽

Chaekyun Kim

Keyword(s):

Hydrogen Peroxide ◽

Raw 264.7 ◽

Taurine Chloramine ◽

Raw 264.7 Macrophages ◽

Induced Apoptosis

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Cornus Officinalis Polyphenol Extract Decrease Pro-inflammatory Markers in Lipopolysaccharide (LPS)-induced RAW 264.7 Macrophages (P06-087-19)

Current Developments in Nutrition ◽

10.1093/cdn/nzz031.p06-087-19 ◽

2019 ◽

Vol 3 (Supplement_1) ◽

Author(s):

Rami Najjar ◽

Neda Akhavan ◽

Shirin Pourafshar ◽

Yun-Hwa Hsieh ◽

Bahram Arjmandi ◽

...

Keyword(s):

Inflammatory Response ◽

Cell Protein ◽

Raw 264.7 ◽

Raw 264.7 Macrophages ◽

Akt Phosphorylation ◽

Downstream Signaling ◽

Funding Sources ◽

Cornus Officinalis ◽

Grant Program ◽

Pre Treatment

Abstract Objectives To investigate whether cornus officinalis (CO) polyphenol extract attenuate the inflammatory response induced by lipopolysaccharide (LPS) in RAW 264.7 macrophages. Methods CO polyphenol extract was prepared using methanolic extraction, followed by solvent evaporation and freeze-drying. RAW 264.7 macrophages were treated with 0, 50, 100, 200 and 400 μg/ml of CO polyphenol extract. After 2 h, cells were then treated with 100 ng/ml of LPS for 6 h. Cells were then collected for whole cell protein expression analysis of signaling and inflammatory molecules via western blot. Results were analyzed using ANOVA followed by Tukey-Kramer post-hoc test. Results LPS treatment significantly increased Akt phosphorylation compared to control (1.00 ± 0.00 vs 0.16 ± 0.02 fold, P < 0.0001). However, pre-treatment with 100, 200 and 400 µg/ml of CO polyphenol extract significantly reduced Akt phosphorylation (0.57 ± 0.08 fold, P = 0.0030; 0.49 ± 0.08 fold, P = 0.0003 and 0.44 ± 0.09 fold, P < 0.0001, respectively) in LPS stimulated macrophages compared to LPS alone. Control cells did not express inducible nitric oxide synthase (iNOS); however, LPS induced iNOS expression, which was significantly decreased by treatment with 400 µg/ml CO polyphenol extract (1.00 ± 0.00 vs 0.36 ± 0.1 fold, P = 0.0098). Similarly, inflammatory cytokines such as interleukin (IL)-1β and 6 were not expressed in control macrophages; however, their expression was induced by LPS. CO polyphenol extract dose-dependently decreased LPS-induced expression of IL-1β (P < 0.0001) and IL-6 (P < 0.0001). Conclusions CO polyphenol extract attenuated the inflammatory response induced by LPS in RAW 264.7 macrophages. This effect is likely due to Akt downstream signaling inhibition. Funding Sources Lewis Foundation Grant Program.

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Wheatgrass extract increases proliferation of RAW 264.7 macrophages induced by hydrogen peroxide (H2O2) or lipopolysaccharide (LPS)

Planta Medica ◽

10.1055/s-0029-1234396 ◽

2009 ◽

Vol 75 (09) ◽

Author(s):

T Özkan ◽

ZA Karabay ◽

A Koç ◽

A Karadag ◽

S Aydos ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Raw 264.7 ◽

Raw 264.7 Macrophages

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Raspberry Polyphenol Extract Decreases NF-kB and IL-6 Expression in Lipopolysaccharide (LPS)-Induced RAW 264.7 Macrophages

Current Developments in Nutrition ◽

10.1093/cdn/nzaa045_055 ◽

2020 ◽

Vol 4 (Supplement_2) ◽

pp. 422-422

Author(s):

Lena Lear ◽

Rami Najjar ◽

Rafaela Feresin ◽

Jessica Danh

Keyword(s):

Alternative Therapy ◽

Control Group ◽

Raw 264.7 ◽

Raw 264.7 Macrophages ◽

Complementary And Alternative Therapy ◽

Funding Sources ◽

Pre Treatment ◽

Post Hoc ◽

Lps Treatment

Abstract Objectives To investigate whether raspberry polyphenol extract attenuates the inflammatory response induced by lipopolysaccharide (LPS) in RAW 264.7 macrophages. Methods Raspberry polyphenol extract was prepared using methanolic extraction, followed by solvent evaporation and freeze-drying. RAW 264.7 macrophages were treated with 0, 50, 100, 200 and 400 μg/ml of raspberry polyphenol extract in six-well plates. After 2 h, cells were then treated with 100 ng/ml of LPS for 6 h. Cells were collected for protein expression analysis of the transcription factor nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and inflammatory cytokines, i.e., interleukin (IL)-6 and IL-1β, via western blot. Results were analyzed using ANOVA followed by Tukey-Kramer post-hoc test. Results As expected, LPS significantly increased NF-kB compared to control (P < 0.0001). Pre-treatment with 400 μg/ml raspberry polyphenol extract significantly decreased phosphorylation of NF-kB in LPS stimulated cells (0.71 ± 0.03 vs. 1.00 ± 0.00-fold, P = 0.005) compared to LPS alone. LPS treatment significantly increased the expression of IL-6 compared to the control group (P < 0.0001). IL-6 expression was significantly reduced in LPS stimulated macrophages by pre-treatment with 200 and 400 µg/ml of raspberry polyphenol extract (0.49 ± 0.03-fold, P < 0.0001 and 0.33 ± 0.04-fold, P < 0.0001 respectively) compared to LPS alone (1.00 ± 0.00-fold). The expression of the inflammatory cytokine IL-1β was significantly greater than control in LPS-only treated cells (P < 0.0001). However, treatment with 400 μg/ml raspberry polyphenol was able to significantly prevent this effect (0.59 ± 0.1 vs. 1.00 ± 0.00-fold, P = 0.007). Conclusions Results indicate that raspberry polyphenols possess anti-inflammatory properties suggesting a possible role as a complementary and alternative therapy to prevent inflammation. However, in vivo and human studies are needed to confirm this. Funding Sources None.

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