[논문]LPS로 유도된 마우스 대식세포주인 RAW264.7에서 MAPK 조절에 의한 백미 물추출물의 항염증 활성

이상호; 유지현; 길기정

doi:10.6116/kjh.2022.37.6.19.

[국내논문] LPS로 유도된 마우스 대식세포주인 RAW264.7에서 MAPK 조절에 의한 백미 물추출물의 항염증 활성
Anti-inflammatory Activity of Cynanchi Atrati Radix Et Rhizoma Water Extracts via Regulation of MAPK in LPS-induced Murine Macrophage Cell Line, RAW 264.7 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.37 no.6, 2022년, pp.19 - 28

이상호 ((주)뉴트렉스테크놀러지) , 유지현 (중부대학교 한방보건제약학과) , 길기정 (중부대학교 한방보건제약학과)

Abstract ▼ AI-Helper

Objectives : To develop natural ingredients that help prevent or treat anti-inflammatory-related diseases and use themas basic data, we investigated anti-inflammatory activity of Cynanchi Atrati Radix Et Rhizoma water extracts(CWE) in lipopolysaccharide(LPS)-induced murine macrophage cell line, RAW 264.7 cells. Methods : The cell viabilities were evaluated with RAW 264.7 cells. The production of nitric oxide(NO), prostaglandin E2(PGE2), pro-inflammatory cytokines such tumor necrotic factor(TNF)-α and interleukin(IL)-6 were assessed in LPS-induced RAW 264.7 cell treated with CWE. Furthermore, the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) were assessed by western blotting. Results : In RAW 264.7 cell, the cell viability by CWE treatment was more than 98.4% at a concentration of 100-400 ㎍/mL. At a concentration of 800 ug/ml of CWE, the cell viability was as low as 86%. At doses of 100, 200 and 400 ㎍/mL, CWE inhibited the production of NO, PGE2, TNF-𝛼 and IL-6 in a dose-dependent manner and also decreased the expression of iNOS and COX-2 from LPS-induced RAW 264.7 cells. In addition, CWE significantly inhibited the MAPK pathway including decreased the phosphorylation of the p38, c-Jun N-terminal kinase(JNK) and extracellular signal-regulated kinase(ERK1/2). Conclusions : Our study provides evidence that CWE inhibits the production of main pro-inflammatory molecules in LPS-induced RAW 264.7 cells via expression of p38, JNK, and ERK1/2 MAPK signaling pathways. Therefore, CWE is expected to be widely used as a natural ingredient for anti-inflammatory functional foods or pharmaceuticals in the future.

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표/그림 (6)

그림 Fig. 1. Effect of CWE on cell viability in murine macrophage cell line, RAW 264.7 cell. Cell viabilities were measured with CCK-8 assay. RAW 264.7 cells were pre-treated with various concentrations of CWE for 1 h, and then LPS(1 ㎍/mL) was treated for 24 h. The concentration of CWE 100, 200, 400 and 800 ㎍ /mL. All values are means±standard deviations of triplicate determinations. Means with different letters(a-b) indicate significant difference(p<0.05) by Duncan's multiple range test.
그림 Fig. 2. Effect of CWE on NO in LPS-induced RAW 264.7 macrophage cells. RAW 264.7 cells were pre-treated with various concentrations of CWE for 1 h, and then LPS(1 ㎍/mL) was treated for 24 h. The concentration of CWE was 100, 200 and 400 ㎍/mL. All values are means±standard deviations of triplicate determinations. Means with different letters(a-e) indicate significant difference(p<0.05) by Duncan's multiple range test.
그림 Fig. 3. Effect of CWE on NO in LPS-induced murine macrophage cell line, RAW 264.7 cell. RAW 264.7 cells were pre-treated with various concentrations of CWE for 1 h, and then LPS(1 ㎍/mL) was treated for 24 h. The concentration of CWE was 100, 200 and 400 ㎍/mL. All values are means±standard deviations of triplicate determinations. Means with different letters(a-e) indicate significant difference(p<0.05) by Duncan's multiple range test.
그림 Fig. 4. Effects of CWE on TNF-α and IL-6 production in LPS-induced murine macrophage cell line, RAW 264.7 cell. RAW 264.7 cells were pre-treated with various concentrations of CWE for 1 h, and then LPS(1 ㎍/mL) was treated for 24 h. The concentration of CWE 100, 200 and 400 ㎍/mL. All values are means±standard deviations of triplicate determinations. Means with different letters(a-e) indicate significant difference(p<0.05) by Duncan's multiple range test.
그림 Fig. 5. Effect of CWE on iNOS and COX-2 protein expression in LPS-induced murine macrophage cell line, RAW 264.7 cell. RAW 264.7 cells were pre-treated with various concentrations of CWE for 1 h, and then LPS(1 ㎍/mL) was treated for 24 h. The concentration of CWE was 100, 200 and 400 ㎍/mL. β-actin was used as an internal control. All values are means±standard deviations of triplicate determinations. Means with different letters(a-d) indicate significant difference(p<0.05) by Duncan's multiple range test.
그림 Fig. 6. Effect of CWE on MAPK singnal expression in LPS-induced murine macrophage cell line, RAW 264.7 cell. RAW 264.7 cells were co-incubated with the indicated concentrations(100, 200 and 400 ㎍/mL) of CWE and LPS for 30 min and whole-cell lysates were analysed by western blotting using antibodies against p38, JNK, and ERK(1/2), and the respective phosphorylated forms. All values are means±standard deviations of triplicate determinations. Means with different letters(a-e) indicate significant difference(p<0.05) by Duncan's multiple range test.

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