[논문]육총용(肉蓯蓉) 열수추출물이 RAW 264.7 대식세포에 미치는 항염증 효과

박선명; 이규영; 홍철희

doi:10.6114/jkood.2022.35.1.011

육총용(肉蓯蓉) 열수추출물이 RAW 264.7 대식세포에 미치는 항염증 효과
Anti-inflammatory Effects of Cistanche deserticola Water Extracts in LPS-stimulated RAW 264.7 Macrophages 원문보기

韓方眼耳鼻咽喉皮膚科學會誌 = The journal of Korean Medicine Ophthalmology & Otolaryngology & Dermatology, v.35 no.1, 2022년, pp.11 - 23

박선명 (상지대학교 한의과대학 안이비인후피부과학교실) , 이규영 (상지대학교 한의과대학 안이비인후피부과학교실) , 홍철희 (상지대학교 한의과대학 안이비인후피부과학교실)

Abstract ▼ AI-Helper

Objectives : The purpose of this study is to confirm the anti-inflammatory effects of Cistanche deserticola Water Extracts(CDWE). Methods : In this study, cell viability was determined by MTT assay. NO production was determined with Griess reagent, and IL-6 production was determined by ELISA. And western blot analysis was performed to confirm the protein expression of NOS2, COX-2, I𝜅B-𝛼, p-I𝜅B-𝛼, and NF-𝜅B. Results : CDWE was not cytotoxic at 15.625-1,000㎍/㎖ in RAW 264.7 cells. CDWE inhibited NO production in a concentration-dependent manner, with inhibitory effect on expression levels of IL-6. Expression level of NOS2, NF-𝜅B p65, and p-I𝜅B-𝛼 decreased at 250-1,000㎍/㎖. mRNA expression of COX-2 suppressed at 250, 1,000㎍/㎖. Conclusions : These results suggest that CDWE has anti-inflammatory effects and can be used as an anti-inflammatory therapeutic material.

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표/그림 (7)

그림 Fig. 1. Effects of CDWE on the Cell Viability in RAW 264.7 Macrophages. RAW 264.7 macrophages were treated with different concentration of CDWE for 24hrs, and cell viability was determined using MTT assay. The values are represented as mean±SD of three independent experiments. CDWE : Cistanche deserticola Water Extracts
그림 Fig. 2. Effects of CDWE on NO Production in LPS-stimulated RAW 264.7 Macrophages. Cells were treated with different concentration of CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 48hrs. NO production was determined with griess reagent. NIL(20μM) was used as a positive control inhibitor. The data shown represent the mean±SD of three independent experiments. ^###p<0.001 vs the control group; ^***p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extracts, NIL : N6-(1-Iminoethyl)lysin
그림 Fig. 3. Effects of CDWE on IL-6 Production in LPS-stimulated RAW 264.7 Macrophages. The production of IL-6 was determined by ELISA. RAW 264.7 macrophages were treated with 250, 500, and 1,000㎍/㎖ CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 24hrs. The data are represented as mean±SD of three independent experiments. ^###p<0.001 vs the control group; ^***p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extract
그림 Fig. 4. Effects of CDWE on NOS2 Protein Expression in LPS-stimulated RAW 264.7 Macrophages. Cells were treated with 250, 500, and 1,000㎍/㎖ CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 24hrs. The protein expression of NOS2 was determined by Western blot analysis using specific antibodies. β–actin was used as internal controls. Relative density was performed using Bio-Rad Quantity One software. The data are represented as mean±SD of three independent experiments. ### p<0.001 vs the control group; ^*p<0.05, ^**p<0.01, ^***p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extract
그림 Fig. 5. Effects of CDWE on COX-2 Protein Expression in LPS-stimulated RAW 264.7 Macrophages. Cells were treated with 250, 500, and 1,000㎍/㎖ CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 24hrs. The protein expression of COX-2 was determined by Western blot analysis using specific antibodies. β–actin was used as internal controls. Relative density was performed using Bio-Rad Quantity One software. The data are represented as mean±SD of three independent experiments. ###p<0.001 vs the control group; ^*p<0.05, ^**p<0.01, ^***p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extract
그림 Fig. 6. Effects of CDWE on the Phosphorylation of IκB-α Protein Expression in LPS-stimulated RAW 264.7 Macrophages. Cells were treated with 250, 500, and 1,000㎍/㎖ CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 20min. The protein expression of IκB-α, p-IκB-α was determined by Western blot analysis using specific antibodies. β–actin was used as internal controls. Relative density was performed using Bio-Rad Quantity One software. The data are represented as mean±SD of three independent experiments. ^###p<0.001 vs the control group; ^***p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extract
$Fig. 7. Effects of CDWE on the NF-κB Activation in LPS-stimulated RAW 264.7 Macrophages. Cells were pre-treated with CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 30min. Nuclear(N) and cytosolic(C) extracts were isolated, and the expression of p65 in each fraction were determined by Western blot analysis. PARP and β–actin were used as internal controls. Relative density was performed using Bio-Rad Quantity One software. The data are represented as mean±SD of three independent experiments. <sup>###</sup>p<0.001 vs the control group; <sup>***</sup>p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extract, PARP : Poly (ADP-ribose) polymerase$ 그림 Fig. 7. Effects of CDWE on the NF-κB Activation in LPS-stimulated RAW 264.7 Macrophages. Cells were pre-treated with CDWE for 1hr prior to the addition of LPS(1㎍/㎖), and the cells were further incubated for 30min. Nuclear(N) and cytosolic(C) extracts were isolated, and the expression of p65 in each fraction were determined by Western blot analysis. PARP and β–actin were used as internal controls. Relative density was performed using Bio-Rad Quantity One software. The data are represented as mean±SD of three independent experiments. ^###p<0.001 vs the control group; ^***p<0.001 vs the LPS-treated group. CDWE : Cistanche deserticola Water Extract, PARP : Poly (ADP-ribose) polymerase

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