Journal List > J Nutr Health > v.52(2) > 1122024

J Nutr Health. 2019 Apr;52(2):129-138. Korean.
Published online Apr 30, 2019.  https://doi.org/10.4163/jnh.2019.52.2.129
© 2019 The Korean Nutrition Society
Anti-inflammatory effects of fruit and leaf extracts of Lycium barbarum in lipopolysaccharide-stimulated RAW264.7 cells and animal model
Su-Mi Bae,1 Ji-Eun Kim,1 Eun-Young Bae,1,2 Kyung-Ah Kim,1 and Sun Yung Ly1,2
1Department of Food and Nutrition, Chungnam National University, Daejeon 34134, Korea.
2Convergence Research Center for Natural Products, Chungnam National University, Daejeon 34134, Korea.

To whom correspondence should be addressed. tel: +82-42-821-6838, Email: sunly@cnu.ac.kr
Received Nov 26, 2018; Revised Jan 31, 2019; Accepted Feb 12, 2019.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.


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.

Keywords: Lycium barbarum; chlorophyll removal; anti-inflammation

Figures


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).
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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).
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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).
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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.
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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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Tables


Table 1
Body weight of mice before and after oral administration
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Table 2
Levels of DNA damage expressed as tail DNA, tail length and tail moment in BALB/c mice lymphocyte
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Notes

This study was supported by grants from the Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology, Republic of Korea (NRF-2017R1D1A3B03028628).

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