[논문]Anti-inflammatory Effect of LFR on LPS-stimulated THP-1 Cells

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Anti-inflammatory Effect of LFR on LPS-stimulated THP-1 Cells 원문보기

대한한방내과학회지 = The journal of internal Korean medicine, v.30 no.2, 2009년, pp.388 - 398

, , , , 정승기 (경희의료원 부속한방병원 한방5내과 의사실)

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Background and Objective: Luffae Fructus Retinervus (LFR) is used for investigating symptoms of inflammation. We have evaluated the anti-inflammatory effect of LFR by analyzing the expression of pro-inflammatory cytokines. Materials and Methods : We differentiated THP-l cells into macrophage-like cells by treatment with PMA. Inflammation was induced by treatment with LPS and PMA. We determined the safe concentration of LFR by using the MTS and MTT assays and using PD 98059 as a negative control for comparison of the anti-inflammatory effect of LFR. Results : The MTS and MTT analysis showed that the cell survival rate was >80% within the LFR concentration range of 10-100 ng/ml and began to decrease to >80% at 1 ${\mu}g/ml$. By RT-PCR analysis, the gene expression of TNF-${\alpha}$, IL-8, TGF-${\beta}$, IL-6, IL-${\beta}$1, and IL-10 levels were down-regulated when monocyte-derived macrophages were treated with concentrations of LFR between 10 ng/mL and 100 ng/mL. Conclusion : We conclude that LFR exerts an anti-inflammatory effect by inhibiting the expression of pro-inflammatory activity. The results suggest a promising way to treat general inflammatory diseases.

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문제 정의

According to the specific needs, tissue macrophages proliferate, further differentiate into more specialized macrophage populations, or become activated¹⁶. In this study, we demonstrated the possibility of herbal medicine as an alternative treatment for inflammatory-related diseases and our research is focused on treatment for the inflammation to investigate LFR extract anti-inflammatory properties.
This experiment studied the expression of cytokines by RT-PCR methods for the analysis of the level of mRNA. For analys is ofthe anti-inflammatory effect of the LFR extract, we determined the expression of inflammation-related cytokine mRNA by RT-PCR analysis.
This study examined the expression of proinflammatory cytokines to control the inflammation. For the analysis of anti-inflammatory effect in terms of the concentration of the LFR extract, the transcriptional activities of IL-1β, IL-6, IL-8, TNF-α, TGF-β, and IL-10 were down-regulated more than the LPS-treated group (Figs.

제안 방법

For the analysis of anti-inflammatory effect in terms of the concentration of the LFR extract, the transcriptional activities of IL-1β, IL-6, IL-8, TNF-α, TGF-β, and IL-10 were down-regulated more than the LPS-treated group (Figs. 3-8).
Given the above background, we administered an extract of LFR to experimentally demonstrate the anti-inflammatory effectiveness of LFR in macrophages in which inflammation was induced by LPS. Then, we measured and analyzed changes in representative pro-inflammatory cytokines, such as IL-1β, IL-6, IL-8, and tumor necros is factor (TNF)-α, and pro-fibrotic cytokines, such as transforming growth factor (TGF)-β, and anti-inflammatory cytokines, such as IL-10, using a reverse transcriptase polymerase chain reaction (RT-PCR).
In this study, we demonstrated the close relationship between LPS-induced inflammation in THP-1 cells and production of inflammatory cytokines by PMA-treated THP-1 cells in LFR extract treatments. The endotoxin (lipopolysaccharide[LPS]) of gram-negative bacteria is believed to be responsible for initiating host responses leading to inflammation.
This study had an emphasison THP-1 cells because the cells are a well-characterized human monocytic leukemic cell line; the cells resemble monocytes with respect to several criteria and can be differentiated into macrophage-like cells by treatment with PMA. PMA-induced differentiation is associated with alterations in cellmorphology, adherence, and expression of severalgenes¹².
To examine the effect of LFR on cell survival, cell viability was analyzed.

대상 데이터

The human monocytic leukemia cell line, THP-1, was obtained from the Korean Cell Line Bank (KCLB; Seoul, Korea). Cells were prepared and resuspended in RPMI 1640 medium (Gibco, USA) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS; Gibco), and 1% antibacterial antifungal solution (Gibco).

데이터처리

Data were presented as the mean ±SEM. Statistical analysis was performed using one-way ANOVA. ***p<0.
The values are expressed as the mean ±S.E.M. The differences between the groups were determined by one-way ANOVA, followed by Tukey’s HSD test for all pairwise multiple comparisons.

이론/모형

Cellviability was measured using the MTS and MTT assays. As a result, macrophages, which were differentiated into white blood cells had a cell viability of >80% with LFR (10 ng/ml).

성능/효과

Finally, the transcriptional activity of IL-10 was down-regulated to 25% and 43% against the LPS treated group. In conclusion, all the cytokine levels were down-regulated by the LFR extract (concentrations up to 100 ng/ml) on monocyte-derived macrophages (Figs. 3-8), which indicates the LFR extract is involved in the inflammatory mechanism(s).

참고문헌 (18)

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Anti-inflammatory Effect of LFR on LPS-stimulated THP-1 Cells 원문보기

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Anti-inflammatory Effect of LFR on LPS-stimulated THP-1 Cells 원문보기

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