[논문]당화된 레스베라트롤의 대식세포 RAW 264.7세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가

라메스 프라시드 판데이; 이지선; 박용일; 송재경

doi:10.4014/mbl.1611.11001

당화된 레스베라트롤의 대식세포 RAW 264.7세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가
Glucosylation of Resveratrol Improves its Immunomodulating Activity and the Viability of Murine Macrophage RAW 264.7 Cells 원문보기

Microbiology and biotechnology letters = 한국미생물·생명공학회지, v.45 no.1, 2017년, pp.19 - 26

라메스 프라시드 판데이 (선문대학교 BT융합 제약공학과) , 이지선 (가톨릭대학교 생명공학과) , 박용일 (가톨릭대학교 생명공학과) , 송재경 (선문대학교 BT융합 제약공학과)

초록
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레스베라트롤의 면역제어 성질과 대식세포의 생존능력과 관련하여 당화된 레스베라트롤의 효과를 확인하기 위해 대식세포 RAW 264.7에서 연구하였다. 인비토로에서 대식세포에서 총 4개의 레스베라트롤 및 당화된 유도체 (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), 및 (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G)를 여러 가지 농도로 처리한 후 일산화질소 (NO)와 인터루킨 6 (IL-6) 발현을 연구하였다. 앞서 언급한 물질로 처리한 후 인비토로에서 RAW 264.7 세포의 생존능력도 연구하였다. 대식세포 생존능력 평가분석 결과를 보면, 두 개의 레스베라트롤 모노글루코사이드인 R-3-G와 R-4'-G은 (E)-resveratrol와 비교하여 A549 and HepG2 세포에서 50-80% 감소된 독성을 보여준다. 당이 없는 레스베라트롤과 비교하면, 당화된 레스베라트 유도체는 긍정적으로 전사적으로 IL-6 및 iNOS 발현이 높아지는 방향으로 NO 및 대식세포에서 IL-6의 생산이 조절된다. 레스베라트롤의 당의 역할은 RAW 264.7 세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가시켜 주는 것으로 보여주고 있다.

Abstract ▼ AI-Helper

Effects of resveratrol glucosylation on the immunomodulation properties of resveratrol and on the viability of macrophage cells have been studied by using murine macrophage RAW 264.7 cells. Nitric oxide (NO) and interleukin 6 (IL-6) expression in macrophages in vitro were studied after treatment with different concentrations of (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), or (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G). In vitro viability of RAW 264.7 cells after treatment with the aforementioned three compounds was also studied. As demonstrated by macrophage cell viability assays, two different resveratrol monoglucosides, R-3-G and R-4'-G, exhibited 50-80% reduced cytotoxicity in comparison to (E)-resveratrol in A549 and HepG2 cells. Compared to the resveratrol aglycon, both glucosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophages positively via transcriptionally up-regulating IL-6 and iNOS expression. Conjugation of a glucose moiety on resveratrol was found to enhance the immunomodulating activity of resveratrol and the viability of RAW 264.7 cells.

주제어

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가설 설정

2D). Considering the effect of resveratrol on cell viability (Fig. 2A), we hypothesize that the dose-dependent decrease in IL-6 and NO levels is due to RAW 264.7 cell cytotoxicity. For the glucosylated derivatives, these were more effective at expressing iNOS and producing NO at relatively higher concentrations [e.

제안 방법

In this study, glucosylation of resveratrol reduced the cytotoxicity of resveratrol, while glycosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophage cells compared to aglycon resveratrol via transcriptionally upregulatiing IL-6 and iNOS expression.
In this study, we studied and compared the effects of (E)-resveratrol and its glucosylated derivatives, R-3-G and R-4'-G, on cell viability and immunomodulating activities using RAW 264.7 cells.
RT-PCR was performed using the ONESTEP RT-PCR PreMix kit (Intron Biotechnology, Korea) with sense and antisense primers for β-Actin (β-Actin, 5'-TGGAATCCTGTGGCATCCATGAAAC-3' (sense) and 5'-TAAAACGCAGCTCAGTAACAGTCCG-3') (antisense));iNOS (5'-CTGCAGCACTTGGATCAGGAACCTG-3' (sense) and 5'-GGGAGTAGCCTGTGTGCACCTGGA-3' (antisense)); and IL-6 (5'-CTGGAGTCACAGAAGGAGTGGC-3' (sense) and 5'-GGCATAACGCACTAGGTTTGCCG-3' (antisense)), under incubation conditions of 95℃ predenaturation for 5 min and 35 cycles of ‘95℃ denaturation for 30 s, 55℃ annealing for 30 s, 72℃ extension for 40 s,’ and a final elongation period of 10 min at 72℃.

대상 데이터

The murine macrophage cell line (RAW 264.7, KCLB # 40071) and several cell lines such as A549 (human lung carcinoma, KCLB # 10185), PC-3 (human prostate carcinoma, KCLB # 21435), HepG2 (human hepatoma carcinoma, KCLB # 88065) HeLa (human cervical carcinoma, KCLB # 10002) and HT-29 (human colon carcinoma, KCLB # 30038) were purchased from the Korea Cell Line Bank (Seoul, Korea) and grown in RPMI-1640 medium (Gibco-Invitrogen, USA) supplemented with 10% (v/v) fetal bovine serum (FBS), 2 mM L-glutamine and 100 U/ml penicillin/streptomycin (PEST). Unless stated otherwise, the cells were incubated with (E)-resveratrol, R-3-G or R-4'-G (50−150 μg/ml) for 24 h.

데이터처리

Comparisons between two groups were made using unpaired Student’s t-tests.
Comparisons between two groups were performed with unpaired Student’s t-tests.

성능/효과

The results demonstrated that glucosylation of resveratrol at either 3 or 4'-position significantly abolished the cytotoxicity of resveratrol on macrophage cells and that glucosylation at 4'-position was shown to be more effective than 3-position in reducing the cytotoxicity of resveratrol.

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