J Nutr Health.  2019 Apr;52(2):129-138. 10.4163/jnh.2019.52.2.129.

Anti-inflammatory effects of fruit and leaf extracts of Lycium barbarum in lipopolysaccharide-stimulated RAW264.7 cells and animal model

• Bae SM
• Kim JE
• Bae EY
• Kim KA
• Ly SY

• KMID: 2444936
• DOI: http://doi.org/10.4163/jnh.2019.52.2.129

Abstract

PURPOSE
Medicinal herbs have recently attracted attention as health beneficial foods and source materials for drug development. Recent studies have demonstrated that extracts of Lycium's fruits and roots have a range of physiologically active substances. The extract of Lycium's leaves has been reported to have excellent anti-oxidant and anti-microbial activity, but its anti-inflammatory efficacy is not known. The chlorophyll present in the leaves can act as an anti-oxidant or pro-oxidant depending on the presence of light. Therefore, this study analyzed the anti-inflammatory effects of Lycium's fruit extract (LFE), leaf extract (LLE), and leaf extract with chlorophyll removal (LLE with CR).
METHODS
This study examined the inhibitory effects of LFE, LLE, and LLE with CR on pro-inflammatory mediator production as well as on the expression of iNOS and COX-2 in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and BALB/c mice.
RESULTS
LFE, LLE, and LLE with CR inhibited the production of pro-inflammatory mediators (NO, TNF-α, IL-6, and IL-1β) and the expression of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells in a dose-dependent manner. Furthermore, the administration of LLE and LLE with CR inhibited the serum pro-inflammatory cytokine levels and suppressed DNA damage in BALB/c mice. In particular, LLE with CR exhibited the highest anti-inflammatory activity.
CONCLUSION
These results suggest that the fruit and leaves of Lycium are potential therapeutic agents against inflammation.

Keyword

Lycium barbarum; chlorophyll removal; anti-inflammation

MeSH Terms

Animals*
Chlorophyll
DNA Damage
Fruit*
Inflammation
Interleukin-6
Lycium*
Mice
Models, Animal*
Plants, Medicinal
RAW 264.7 Cells
Chlorophyll
Interleukin-6

Figure

• 

  Fig. 1 Effect of Lycium's fruit extracts (LFE) and leaf extracts (LLE), leaf extracts chlorophyll removal (LLE with CR) on cell viability of lipopolysaccharide (LPS)-induced RAW264.7 cells. Cells were treated with LFE, LLE, LLE with CR (31.25, 62.5, 125, 250, 500, 1,000 μg/mL), then with or without LPS (1 μg/mL) for 24 h. The cell proliferation was estimated by the MTT assay with WST system. Each bar represents the mean ± SD. NS: not significantly different by Duncan's multiple range test (p < 0.05).

• 

  Fig. 2 Effect of Lycium's fruit extracts (LFE) and leaf extracts (LLE), leaf extracts chlorophyll removal (LLE with CR) on the production of nitric oxide (NO) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Cells were treated with LFE, LLE, LLE with CR (31.25, 62.5, 125, 250, 500, 1,000 μg/mL), then with or without LPS (1 μg/mL) for 24 h. The culture supernatant of the treated cells were used to measure NO level. Levels of nitric oxide were determined by Griess reagent. Each bar represents the mean ± SD. Significant values are represented by an asterisk (∗) (p < 0.05 compared to the group treated with LPS alone).

• 

  Fig. 3 Effect of Lycium's fruit extracts (LFE) and leaf extracts (LLE), leaf extracts chlorophyll removal (LLE with CR) on the production of TNF-α (A), IL-6 (B), IL-1β (C) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Cells were treated with LFE, LLE, LLE with CR (31.25, 62.5, 125, 250, 500, 1,000 μg/mL), then with or without LPS (1 μg/mL) for 24 h. The levels of pro-inflammatory cytokines in the cell culture supernatant were determined by ELISA. Each bar represents the mean ± SD. Significant values are represented by an asterisk (∗) (p < 0.05 compared to the group treated with LPS alone).

• 

  Fig. 4 Effect of Lycium's fruit extracts (LFE) and leaf extracts (LLE), leaf extracts chlorophyll removal (LLE with CR) on iNOS and COX-2 protein expression in lipopolysaccharide (LPS)-induced RAW264.7 cells. Cells were treated with LFE, LLE, LLE with CR (125, 250, 500 and 1,000 μg/mL), then with or without LPS (1 μg/mL) for 24 h. Cell lysates were used for western blot analysis. The levels of protein expression in iNOS and COX-2 were normalized to the β-actin signals. The relative band intensities are indicated above each band.

• 

  Fig. 5 Effect of Lycium's leaf extracts (LLE), leaf extracts chlorophyll removal (LLE with CR) on the production of serum TNF-α (A), IL-6 (B), IL-1β (C) in LPS-induced BALB/c mice. LLE and LLE with CR (200 mg/kg body weight) was administered orally in mice for seven days, and then challenged intraperitoneally with LPS (5 mg/kg body weight). The levels of pro-inflammatory cytokines in the serum were determined by ELISA. Serum TNF-α, IL-6, IL-1β levels were analyzed 8 h after the last LPS challenge. Each bar represents the mean ± SD. Significant values are represented by an asterisk (∗) (p < 0.05 compared to the group treated with LPS alone).

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