[논문]NF-κB와 MAPK억제를 통한 시호소간탕(柴胡疏肝湯)의 항염증효과

진효정; 박상미; 김예림; 변성희; 김상찬

doi:10.14374/hfs.2023.31.2.99

NF-κB와 MAPK억제를 통한 시호소간탕(柴胡疏肝湯)의 항염증효과
Anti-inflammatory effect of Sihosogan-tang via inhibition of NF-κB and MAPK cascade 원문보기

大韓韓醫學方劑學會誌 = Herbal formula science, v.31 no.2, 2023년, pp.99 - 109

진효정 (대구한의대학교 한의과대학 방제학교실) , 박상미 (대구한의대학교 한의과대학 방제학교실) , 김예림 (대구한의대학교 한의과대학 방제학교실) , 변성희 (대구한의대학교 한의과대학 방제학교실) , 김상찬 (대구한의대학교 한의과대학 방제학교실)

Abstract ▼ AI-Helper

Objectives : Sihosogan-tang (SST) is one of the traditional herbal formula and also one of the Korean medical insurance medicines. It commonly used in the treatment of hepatitis, chronic gastritis, intercostal neuralgia, pleurisy, and depression in East Asia. In the present study, we have demonstrated the anti-inflammatory effects of SST in macrophage cell line. Methods : To investigate mechanism of the anti-inflammatory effect of SST, we examined the productions of nitric oxide (NO) and pro-inflammatory cytokines, and the expressions of inducible NO synthase (iNOS), nuclear factor-κ B (NF-κB) and mitogen-activated protein kinase (MAPK) on RAW 264.7 cells activated by LPS. Results : SST significantly inhibited the expression of iNOS increased by LPS, and also significantly inhibited the production of NO. In addition, SST significantly inhibited pro-inflammatory cytokines such as TNF- α and interleukines. SST inhibited the expression of NF-κB and MAPK activation. Conclusions : These results suggest that SST ameliorates inflammatory response in LPS-activated RAW 264.7 cells through the inhibition of the NF-κB and MAPK pathway. Therefore, this study supplies objective evidence for the anti-inflammatory effect of SST.

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표/그림 (5)

그림 Fig. 1. The inhibitory effects of SST on LPS-mediated NO production. RAW 264.7 cells were treated with 30-1000 μg/mL SST for 24 hours, and cell viability was measured to investigate the cytotoxicity of SST (A). Cell viability of SST plus LPS (2 μg/mL) was measured by MTT assay (B). To measure NO production, Griess reagent assay was performed with the culture media of cells treated with SST 30-300 μg/mL, (1 h) and LPS (2 μg/mL, 18 h) (C). Values represent the mean ± S.D. of three independent experiments for each condition (significant compared to the untreated control, **P < 0.01; significant as compared to LPS alone, ##P < 0.01; N.S., not significant).
그림 Fig. 2. The inhibitory effects of SST on LPS-mediated iNOS expressions. To confirm the expression of the iNOS protein, RAW 264.7 cells were pretreated with 100, 300 μ g/mL of SST of 1 h before LPS (2 μg/mL, 18 h) treatment. Equal amounts of protein were separated by SDS-PAGE and β-actin was used as an immunoblot loading control (A). The iNOS intensity was determined by scanning densitometry (B). Values represent the mean ± S.D. of three independent experiments for each condition (significant compared to the untreated control, **P < 0.01; significant as compared to LPS alone, ##P < 0.01).
그림 Fig. 3. The inhibitory effects of SST on LPS-mediated pro-inflammatory cytokines production. Raw 264.7 cells were pretreated with 100, 300 μg/mL of SST for 1 h and further incubated with LPS (2 μg/mL) for 18 h. The concentrations of IL-1β (A), IL-6 (B) and TNF-α (C) in culture media were evaluated by ELISA kits as described in materials and methods part. Values represent the mean ± S.D. of three independent experiments for each condition (significant compared to the untreated control, **P < 0.01; significant as compared to LPS alone, ##P < 0.01).
$Fig. 4. The inhibitory effects of SST on LPS-mediated p-IκBα and NF-κB expressions. RAW 264.7 cells were pretreated with 100, 300 μg/mL of SST of 1 h before LPS (2 μg/mL) treatment. The levels of (A) IκBα (15min in the cytoplasmic fraction), (B) p-IκBα (30min in the cytoplasmic fraction) and (C) NF-κB (30min in the nuclear fraction) were monitored by scanning densitometry. β-actin and lamin A/C were used as an immunoblot loading controls. Values represent the mean ± S.D. of three independent experiments for each condition (significant compared to the untreated control, **P < 0.01; significant as compared to LPS alone, ##P < 0.01, #P < 0.05).$ 그림 Fig. 4. The inhibitory effects of SST on LPS-mediated p-IκBα and NF-κB expressions. RAW 264.7 cells were pretreated with 100, 300 μg/mL of SST of 1 h before LPS (2 μg/mL) treatment. The levels of (A) IκBα (15min in the cytoplasmic fraction), (B) p-IκBα (30min in the cytoplasmic fraction) and (C) NF-κB (30min in the nuclear fraction) were monitored by scanning densitometry. β-actin and lamin A/C were used as an immunoblot loading controls. Values represent the mean ± S.D. of three independent experiments for each condition (significant compared to the untreated control, **P < 0.01; significant as compared to LPS alone, ##P < 0.01, #P < 0.05).
그림 Fig. 5. The inhibitory effects of SST on LPS-mediated phosphorylation of MAPK. The cells were pretreated with 300 μg/mL of SST for 1 h and followed by incubated with LPS (2 μ g/mL) for 1 h. Equal amounts of protein were separated by SDS-PAGE. The levels of MAPK expression were determined using specific p-p44/42, p-p38 and p-JNK antibodies. β-actin was used as an immunoblot loading control (A). The phosphorylation of MAPK intensity was determined by scanning densitometry (B). Values represent the mean ± S.D. of three independent experiments for each condition (significant compared to the untreated control, **P < 0.01; significant as compared to LPS alone, ##P < 0.01, #P < 0.05).

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