1.Results of the Study on the Effect of Glycyrrhiza uralensis and Inula helenium L. Extracts on Nitric Oxide Production in a Lipopolysaccharide-Induced Inflammation Model
Anand A ; Ariunzaya Lkh ; Ariunzaya M ; Enkhsaikhan Lkh ; Zolzaya B ; Sarnai Ts ; Shiirevnyamba A ; Ariunzaya B
Mongolian Journal of Health Sciences 2025;86(2):70-76
Background:
Nitric oxide (NO) is a biological messenger molecule that plays a significant role in the pathogenesis of
inflammation. It has anti-inflammatory effects under physiological conditions but can act as a pro-inflammatory mediator
when produced excessively under abnormal conditions. NO is involved in the pathogenesis of inflammatory diseases affecting
the joints, intestines, and lungs. Therefore, compounds that inhibit NO production are considered important for the
treatment of inflammatory diseases and are used clinically. The RAW 264.7 mouse macrophage-like cell line is a widely
used model for inflammation studies. Lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative
bacteria, is used to activate RAW 264.7 cells and create an inflammation model. Glycyrrhiza uralensis, also known as
licorice, is a perennial herbaceous plant in the Fabaceae family. It has been widely used in traditional medicine due to its
anti-inflammatory, antiviral, and hepatoprotective properties. Recent studies have shown that licorice contains bioactive
compounds such as glycyrrhizin, liquiritigenin, and isoliquiritigenin, which play an important role in inhibiting the synthesis
of pro-inflammatory cytokines in macrophages induced by LPS. Inula helenium L., also known as elecampane, is a
perennial herbaceous plant used as an expectorant, anti-infective, anti-inflammatory, and anti-helminthic agent in various
respiratory diseases. Licorice and Inula helenium are included in Mongolian traditional medicine prescriptions, but their
anti-inflammatory effects have not been fully determined, which forms the basis for this research.
Aim:
To study the effect of Glycyrrhiza uralensis and Inula helenium extracts on the production of NO, the end product
of inflammation, in RAW 264.7 macrophage cell lines stimulated with lipopolysaccharide.
Materials and Methods:
The non-toxic dose of the plant extracts was determined in RAW 264.7 mouse macrophage-like
cell line cultures using the MTT assay. Nitric oxide production in RAW 264.7 cell line cultures stimulated with lipopolysaccharide
was assessed using the Griess method. Statistical analysis of the results was performed using SPSS 25.0
software, with the p-value calculated by one-way ANOVA, and the differences between groups were evaluated.
Results:
In RAW 264.7 cell cultures, Glycyrrhiza uralensis and Inula helenium extracts were non-toxic and promoted
cell growth at doses ranging from 1 to 25 μg/ml, while a dose of 50 μg/ml was toxic and inhibited cell growth (p<0.01).
When the combined plant extracts were applied to cells at doses ranging from 1 to 100 μg/ml, they were also non-toxic
and enhanced cell growth, while a dose of 500 μg/ml was toxic and inhibited growth (p<0.001). In terms of nitric oxide
production, Glycyrrhiza uralensis extract increased NO production in a dose- and time-dependent manner compared to
the control or PBS-treated group. However, Inula helenium extract did not show a dose- or time-dependent effect on NO
production. In the lipopolysaccharide-induced inflammation model, licorice extract inhibited NO production at a dose of
30 μg/ml after 12 hours, and further reduced NO production in a dose- and time-dependent manner after 48 hours. Conversely,
no significant changes were observed in the Inula helenium extract group at a dose of 25 μg/ml after 48 hours, but
a reduction in LPS-induced NO production was observed at a dose of 25 μg/ml after 48 hours.
Conclusion
Glycyrrhiza uralensis extract alone increased NO production in a dose- and time-dependent manner. It also
reduced LPS-induced NO production in a dose- and time-dependent manner. In contrast, Inula helenium extract inhibited
LPS-induced NO production at a dose of 25 μg/ml after 48 hours.