Anti-inflammatory Activity of Ethanol Extract of Undaria pinnatifida Root in RAW 264.7 Cells

Kor J Fish Aquat Sci 47(6),751-756,2014

한수지 47(6), 751-756, 2014

Original Article

751

Copyright © 2014 The Korean Society of Fisheries and Aquatic Science pISSN:0374-8111, eISSN:2287-8815

서 론

미역

(Undaria pinnatifida)

은우리나라에서이용되고있는대 표적인갈조류로무기질과비타민등의영양성분을풍부하게 함유하고있으며기능성지방산함량및광물질함량이풍부하 다

(Choi et al., 1992; Kim and Choi, 1994).

^{이러한미역은유} 용한영양성분을많이함유하고있지만

,

^{뿌리와같은부산물들} 은활용되지못하고폐기되는부분이많아바다에버려져환경 오염을유발한다

.

^{바다로폐기되는미역의양은전체미역중}

40-60%

^{로매년남해안에}

14-21

^{만톤이폐기되고있다}

(Ahn et al., 2004).

^{미역을채취할때주로미역의밑부분이바다에버} 려지게되며

,

^{육상에서가공중에도많은부분이폐기물로버} 려지고있는실정이다

.

미역폐기물들은좁쌀병발병원인이되 고양식장의심각한오염과해양오염의가속화를촉진시키나

(Ahn et al., 2004)

^사료

,

^{유기질비료로사용될수있으며}

(Jung,

2013),

^{중금속이온의흡착효과가있는것으로보고되고있다}

(Cho et al., 2004).

미역의생리활성은항염증효과

(Jeong et al., 2012; Choi and Kim, 2013)

^{등다양하게보고되고있지만부} 산물인뿌리의생리활성에대한연구는아직보고된바가없다

.

한편염증반응은외부로부터생체내로박테리아나바이러스 와같은외부항원이침입하여물리화학적손상을입었을때그 손상에대해수복재생하려는조직반응을말한다

(Olefsky and Glass, 2010).

^{면역세포중대식세포는인체면역체계에서중} 요한역할을하는백혈구로써

,

^{모든조직에존재하며외부의자} 극으로인하여활성화되어염증매개물질을분비하여염증반응 을유발한다

(Albina and Reichner, 1995).

^{대식세포는그람음} 성균의세포외막에존재하는

lipopolysaccharide (LPS)

와같은 외부의자극에의해염증반응을조절하는핵심적인전사인자를

RAW 264.7 세포에서의 미역(Undaria pinnatifida) 뿌리 에탄올 추출물의 항염증활성

강보경·안나경·최연욱·김민지

¹

·박시우·박원민·김보람·김꽃봉우리

¹

·안동현*

부경대학교 식품공학과/식품연구소, ¹부경대학교 수산과학연구소

Anti-inflammatory Activity of Ethanol Extract of Undaria pinnatifida Root in RAW 264.7 Cells

Bo-Kyeong Kang, Na-Kyung Ahn, Yeon-Uk Choi, Min-Ji Kim

¹

, Si-Woo Bark, Won-Min Pak, Bo-Ram Kim, Koth-Bong-Woo-Ri Kim

¹

and Dong-Hyun Ahn*

Department of Food Science and Technology/Institute of Food Science, Pukyong National University, Busan 608-737, Korea

1

Institute of Fisheries Sciences, Pukyong National University, Busan 619-911, Korea

The anti-inflammatory effects of the ethanol extract of Undaria pinnatifida root (UPREE) were investigated using the lipopolysaccharide (LPS)-induced inflammatory response in RAW 264.7 cells by measuring the production of nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β, and cell proliferation. We found that NO levels were reduced by 34% at 100 μg/mL. Moreover, the production of IL-6 and TNF-α was suppressed by the UPREE treatment. In particular, the IL-6 production was inhibited by more than 30% at 100 μg/mL UPREE. The pro- liferation of RAW 264.7 cells was measured by MTT assay, and we found no cytotoxicity in those cells treated with UPREE compared to the control. Our results suggest that UPREE shows promise as a therapeutic anti-inflammatory treatment.

Key words: Undaria pinnatifida , Anti-inflammatory Activity, IL-6, TNF-α

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens (http://creativecommons.org/licenses/by-nc/3.0/)which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

http://dx.doi.org/10.5657/KFAS.2014.0751 Kor J Fish Aquat Sci 47(6) 751-756, December 2014

Received 14 August 2014; Revised 22 October 2014; Accepted 17 November 2014

*Corresponding author: Tel: +82. 51. 629. 5831 Fax: +82. 51. 629. 5824

E-mail address: [email protected]

강보경

ㆍ

안나경

ㆍ

최연욱

ㆍ

김민지

ㆍ

박시우

ㆍ

박원민

ㆍ

김보람

ㆍ

김꽃봉우리

ㆍ

안동현

752

활성화시켜

, nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6

등여러가지염증매개인자들이생성 되게된다

(Wu et al., 2004; Valledor et al., 2010).

^{염증매개인} 자들은조직손상의복구와보호에필요한것이지만

(Dinarello,

1998),

^{이러한염증반응이과도하게일어나게되면점막손상을}

촉진하고

,

^암

,

^천식

,

^기관지염

,

^관절염

,

^{파킨스병과같은퇴행성} 뇌질환등의각종질환을유도하기때문에생명현상유지를위 해염증반응조절은매우중요하다

(Willoughby, 1975).

^현재까 지개발되어이용되고있는일부합성염증억제제는간손상

,

위 장관출혈

,

^{성장억제와같은부작용등을초래하여사용시많은} 주의와제한성이있다

.

^{이에많은연구자들은보다안전하고효} 과적인천연물유래의항염증물질을찾는연구를계속하고있 다

(Kwak and Lee, 2014).

^{그러므로폐기되고있는미역뿌리의} 고부가가치기능성소재에대한가능성을알아보고

,

새로운자 원으로활용하기위한연구가필요하다

.

따라서본연구는대량폐기되고있는미역부산물의재활용 방안으로서

,

미역뿌리에탄올추출물이

LPS

로염증을일으켜

활성화된

RAW 264.7 cells

^의

NO, IL-6, TNF-α

^및

IL-1β

^분비 에미치는영향에대해조사하여항염증효과를알아보고천연 항염증제로서이용가능성에대해구명하고자하였다

.

재료 및 방법

실험재료

미역뿌리는부산

(

^주

)

^{석하에서제공 받았으며}

,

^{담수로깨끗} 이수세하고동결건조

(EyelaFDU-2100, Tokyo Rikakikai Co., Tokyo, Japan)

후에이를분말화하고진공포장하여

-20℃

에서 저장하여두고실험에사용하였다

.

추출

분쇄한미역뿌리분말에

10

^배의

95%

^{에탄올을가하여추출} 한후원심분리기

(UNION 32R, Hanil Co., Incheon, Korea)

^로

3,000 rpm

에서

10

분간원심분리한후상층액을취하였다

.

얻어 진잔사는이와동일한방법으로

2

^{회반복하여추출하였다}

. 3

회추출하여얻어진상층액을

filter paper (Advantec5A, Toyo Roshi Kaisha, Tokyo, Japan)

^{를이용하여여과하였다}

.

^그상층 액을

35℃ water bath

^에서

rotary evaporator (RE200, Yamato Co., Tokyo, Japan)

를이용하여감압하여농축하였다

.

농축물 은

37℃

^{에서건조시키고}

-20℃

^{에보관하여두고실험에사용} 하였다

.

세포배양

마우스의대식세포주인

RAW 264.7

^세포

(KCLB40071)

^는한 국세포주은행에서분양받아사용하였으며

, DMEM (GIBCO, Grand Island, NY, USA)

^에

10% inactivated fetal bovine se- rum (FBS)

와

1% penicillin-streptomycin

을첨가한배지를배

양액으로

37℃, 5% CO

₂

incubator (MCO-15AC, Sanyo, Osa- ka, Japan)

에서배양하였다

.

실험과정의모든세포는

80-90%

정 도의밀도로자랐을때계대배양하였고

, 20 passages

^를넘기지 않은세포만사용하였다

.

세포 독성 측정

시료의 세포독성을 평가하기 위해

3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay

를실시하 였다

. RAW 264.7 cell (1×10

⁶

cells/mL)

^를

96-well plate

^에분 주하고

20

시간전배양후

, 2 μg/mL

의

LPS (Sigma, St. Louis, MO, USA)

^와추출물

(0.1, 1, 10, 50, 100 μg/mL)

^{로첨가하여}

37℃, 5% CO

₂^에서

22

^{시간배양하였다}

.

^배양후

, 5 mg/mL

^농 도의

MTT

시약을첨가하여

2

시간재배양하고이를

4℃, 2,000 rpm

^에서

10

^{분간원심분리하여상층액을제거하였다}

.

^그후

,

^각

well

에

dimethyl sulfoxide (DMSO)

를첨가하고이를

micro- plate reader (Mod el 550, Bio-Rad, Richmond, CA, USA)

^를 이용하여

540 nm

^{에서흡광도를측정하였다}

.

^{세포증식능은다} 음식에의해계산하였다

.

Proliferation Index (%)=sample

^흡광도

/control

^흡광도

×100 NO 분비량 측정

NO

^{의농도는배양액내의}

nitrite

^농도를

griess

^{반응을이용} 하여측정하였다

. RAW 264.7

^세포는

DMEM

^{배지를이용하} 여

2.5×10

⁵

cells/mL

로 조절한 후

24-well plate

에 접종하고

37℃, 5% CO

₂^농도의

incubator

^에서

20

^{시간전배양하였다}

.

^배 지를교환후

, RAW 264.7

세포에

2 μg/mL

의

LPS

와추출물을

(0.1, 1, 10, 50, 100 μg/mL)

^{로처리하여}

24

^{시간재배양하였다}

.

배양액의 상층액을얻은후

,

^동량의

griess

^시약

(1% sulfanil- amide+0.1% naphthylen diamine dihydrochloride, 1：1)

을첨 가하여실온에서

10

^{분간반응시킨후}

, microplate reader

^를이 용하여

540 nm

에서흡광도를측정하였다

.

세포배양액내

NO

의농도는

sodium nitrite (NaNO

₂

)

^{의농도별표준곡선과비교} 하여산출하였다

.

Pro-inflammatory cytokine 분비량 측정

세포배양액내의

IL-6, TNF-α

^및

IL-1β cytokine

^의분비량 을

ELISA-kit (BD Bioscience, San Jose, CA, USA)

를이용하 여측정하였다

. RAW 264.7

^세포는

DMEM

^{배지를이용하여}

2.5×10

⁵

cells/mL

^{로조절한후}

24-well plate

^{에접종하고}

37℃,

5% CO

₂

incubator

에서

18

시간전배양하였다

.

세포에

2 μg/mL

의

LPS

^{와미역뿌리}

ethanol

^{추출물을처리하여}

12

^{시간재배양} 하였다

.

먼저

, ELISA microplate

에

anti-mouse IL-6, TNF-α

및

IL-1β mouse antibody (mAb)

^{를분주하여하룻밤동안}

coat-

ing

^시켰다

.

^이를

0.05% tween 20

^이포함된

phosphate buffered

saline (PBST)

로 세척하고

, 10% FBS

용액으로

blocking

하 였다

. PBST

^{로세척한뒤}

,

^각

microplate

^{에배양상층액을넣} 고실온에서

2

시간반응시켰다

.

다시

PBST

로세척한뒤희석

미역뿌리 에탄올 추출물의 항염증 활성

753

한

biotinylated anti-mouse IL-6, TNF-α mAb

^와

streptavidin- horseradish peroxidase conjugate

^{를첨가하여실온에서}

1

^시간 반응시켰다

. IL-1β

^의경우

, biotinylated anti-mouse IL-1β de- tection antibody

^{를첨가하고}

1

^{시간반응시킨후}

, streptavidin- horseradish peroxidase conjugate

를첨가하여

30

분반응시켰 다

.

^그후

,

^이를다시

PBST

^{로세척한다음}

, ο-phenylenediamine (OPD)

^{용액을첨가하여암반응시키고}

microplate reader

^를이 용하여

490 nm

^{에서흡광도를측정하였다}

.

통계 처리

모든실험결과에대한통계처리는

SAS program (Statistical analytical system V8.2, SAS Institute Inc., Cary, NC, USA)

^을

이용하여

ANOVA

^{법으로분산분석을실시하였으며}

,

^조사항목

들간의유의성검정은

Duncan

^{의다중검정법으로}

P<0.05

수준 에서실시하였다

.

결과 및 고찰

미역뿌리 에탄올 추출물에 의한 세포독성 평가

MTT assay

^{를이용하여미역뿌리에탄올추출물의처리농도}

에따른

RAW 264.7

세포의증식능을측정하였다

.

그결과

,

미 역뿌리에탄올추출물을

0.1, 1, 10, 50, 100 μg/mL

^{의농도로처} 리시모든농도에서무처리구에비해세포증식률이증가함을 보여세포독성이없음을확인하였다

(Fig. 1).

^{따라서미역뿌리} 에탄올추출물의

NO

^및

cytokine

^{분비억제효과가세포독성} 에의한효과가아님을확인하였다

.

미역뿌리 에탄올 추출물에 의한 NO 생성 억제효과 NO

^{는생체내에서}

NO synthase (NOS)

^{라는효소에의해}

L-

arginine

^{으로부터생성되는반응성이강한자유라디칼로써신}

경전달

,

^{면역반응및혈관확장등의생리적인기능조절에중} 요한역할을하는신호전달물질이다

(Weisz et al., 1996; Kim et al., 2012).

^하지만

LPS

^및

pro-inflammatory cytokine

^에의 해다량의

NO

가생성되고

,

과발현된

NO

는혈관투과성을증 가시켜부종을일으키고염증반응을촉진하여조직손상및종 양발생등에관여하게된다

(Stuehr et al., 1991).

^{따라서미역뿌} 리에탄올추출물이

NO

생성량에미치는영향을알아보기위 하여대식세포인

RAW 264.7

^세포에

LPS

^{로염증반응을유도} 시키고미역뿌리에탄올추출물을농도별로

(0.1, 1, 10, 50, 100 μ g/mL)

^{처리하여 세포배양상층액내의}

NO

^{양을측정하였} 다

.

^그결과

, LPS

^에의해약

21

^배증가된

NO

^{생성량이미역뿌} 리에탄올추출물에의해

LPS

처리대조군보다농도별로

2%, 4%, 22%, 32%, 34%

^감소하여

NO

^{생성량이유의적으로감} 소하는것으로나타났으며

(P<0.05), IC

₅₀값은

147 μg/mL

이였 다

(Fig. 2). Choi et al. (2008)

^{은다시마가}

200 μg/mL

^농도에서

NO

^분비량이

21%

^{감소되었다고보고하여미역뿌리가다시마}

보다

NO

분비량억제에더효과가있음을알수있다

.

따라서 미역뿌리에탄올추출물이세포독성없이

NO

^{의생성을감소} 시켜항염증효과에기여함을확인하였다

.

미역뿌리 에탄올 추출물에 의한 RAW 264.7 세포의 IL-6 분비 감소 효과

대식세포인

RAW 264.7

은그람음성세균의세포외막에존 재하는

LPS

^{자극에의해}

IL-6, TNF-α

^및

IL-1β

^와같은

pro- inflammatory cytokine

들의분비가촉진된다

(Guha and Mack- man, 2001). IL-6

^는

T

^세포

,

^{단핵구및대식세포등에의해생성} 되고림프구의기능을활성화시키며생체내의조혈작용을조 절하는급성면역반응의유도물질이다

(Akira et al.,1990; Shan

Fig. 2. Inhibitory effect of ethanol extract of Undaria pinnatifida root on the production of NO in RAW 264.7 cells.

a-eMeans with different superscripts are significantly different (P<0.05).

0 100 200 300

Sample - 0.1 1 10 50 100 (µµg/mL)

a a a a a

b

0 10 20 30 40 50 60

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1 e

a ab b

c

d d

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1 e

a

b bc c

d d

0 500 1,000 1,500 2,000 2,500

c

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

d d

f

a b

e 0

100 200 300

Proliferation index Nitric oxide (µM) IL-6 (pg/mL) TNF-α (pg/mL) IL-1β (pg/mL)

0 20 40 60 80

b

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1

a a

a a a

a

Fig. 1. Inhibitory effect of ethanol extract of Undaria pinnatifida root on the production of proliferation in RAW 264.7 cells.

a-bMeans with different superscripts are significantly different (P<0.05).

0 100 200 300

Sample - 0.1 1 10 50 100 (µµg/mL)

a a a a a

b

0 10 20 30 40 50 60

Sample -LPS +

- + + +

1 10 100 (µµg/mL)

+ 50

- +

0.1 e

a ab b

c

d d

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1 e

a

b bc c

d d

0 500 1,000 1,500 2,000 2,500

c

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

d d

f

a b

e 0

100 200 300

Proliferation index Nitric oxide (µM) IL-6 (pg/mL) TNF-α (pg/mL) IL-1β (pg/mL)

0 20 40 60 80

b

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

a a

a a a

a

강보경

ㆍ

안나경

ㆍ

최연욱

ㆍ

김민지

ㆍ

박시우

ㆍ

박원민

ㆍ

김보람

ㆍ

김꽃봉우리

ㆍ

안동현

754

et al., 2009).

^또한

IL-6

^{는염증을유발하여대식세포를 포함} 한식세포의탐식작용과보체생산을증진시키는기능을한다

(Hur et al., 2013). LPS

^로유도된

RAW 264.7

^{세포에미역뿌}

리에탄올추출물을 처리한후배양액내에서의

IL-6

의 분비

량을확인한결과

, LPS

^{만처리한대조군에서는}

259.50 pg/mL

의분비량을보였으나

,

^{미역뿌리에탄올추출물처리시농도별} 로각각

222.86, 209.77, 200.61, 167.90, 161.35 pg/mL

의분비 량을보여농도의존적으로

IL-6

^{의분비가억제됨을확인하였} 으며

(Fig. 3),

특히

50

및

100 μg/mL

의농도에서는약

35%

와

38%

^{의감소효과를확인하여미역뿌리에탄올추출물이}

RAW

264.7

^세포에서

IL-6

^{발현을억제하여염증억제효과를가짐을} 확인하였다

. Choi et al. (2013)

은미역에탄올추출물로부터분 리한

ethyl acetate

^{분획물의항염증효과에서}

50 μg/mL

^의농도 에서는감소효과가없었으나

100 μg/mL

농도에서는

63%

의염 증억제효과를나타내었다고보고하여

,

^{미역뿌리에탄올추출} 물이저농도에서항염증효과가더큰것으로사료된다

. 미역뿌리 에탄올 추출물에 의한 RAW 264.7 세포의 TNF-α 분비 감소 효과

TNF-α

는염증성매개물질중전구염증

cytokine

중하나로 많은감염성혹은자가면역성질환에서강력한면역매개물 질로여겨지고 있는데

,

^{이는대식세포}

,

^활성화

T

^세포

,

^비만세 포에서생산되며중성구

,

림프구

,

단핵세포를활성화시키는작 용을한다

(Kim et al., 2009).

^{미역뿌리에탄올추출물이염증} 성

cytokine

인

TNF-α

의분비에미치는영향을알아보기위해

LPS

^{로염증반응을유도한후}

,

^{미역뿌리에탄올추출물을처리} 한배양상층액을이용하여

TNF-α

^분비량을

ELISA

^방법으로 측정하였다

. RAW 264.7

세포에서분비된

TNF-α

의양은

LPS

로세포를자극시켰을때에

2,301 pg/mL

^{으로현저히증가하였} 다

.

그러나미역뿌리에탄올추출물을

0.1-100 μg/mL

농도로 처리한세포에서는

LPS

^{로세포를자극시켰을때와비교시}

,

^미

역뿌리에탄올추출물의농도에의존하여유의적으로감소하였 다

(Fig. 4).

특히

100 μg/mL

농도로처리한세포에서는

22%

의 감소를보였다

.

^이는미역

ethyl acetate

^추출물이

100 μg/mL

^농 도에서

TNF-α

분비량을

21%

정도감소시킨것과유사한경향 을보였다

(Choi et al., 2013).

^{이러한결과는}

NO

^및

IL-6

^의분 비감소효과보다는적었지만미역뿌리에탄올추출물이

RAW 264.7

세포에서

TNF-α

의발현을억제하여항염증기능에관여 하고있음을알수있다

.

미역뿌리 에탄올 추출물에 의한 RAW 264.7 세포의 IL-1β 분비 감소 효과

IL-6, TNF-α

^{와 함께 대표적인 염증성}

cytokine

^인

IL-1β

^는

NO

생성의매개물질로대식세포

,

단핵구

, B

세포등에서분비 된다

.

^{또한국소염증을발생시키며}

, T

^{세포의활성화}

, B

^세포의 성숙및

NK cell

을활성화시키는

cytokine

이다

(Lee, 2011).

또

Fig. 3. Inhibitory effect of ethanol extract of Undaria pinnatifida root on the production of IL-6 in RAW 264.7 cells.

a-eMeans with different superscripts are significantly different (P<0.05).

0 100 200 300

Sample - 0.1 1 10 50 100 (µµg/mL)

a a a a a

b

0 10 20 30 40 50 60

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1 e

a ab b

c

d d

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1 e

a

b bc c

d d

0 500 1,000 1,500 2,000 2,500

c

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL) - + 50

0.1

d d

f

a b

e 0

100 200 300

Proliferation indexNitric oxide (µM)IL-6 (pg/mL)TNF-α (pg/mL)IL-1β (pg/mL)

0 20 40 60 80

b

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

a a

a a a

a

Fig. 4. Inhibitory effect of ethanol extract of Undaria pinnatifida root on the production of TNF-α in RAW 264.7 cells.

a-fMeans with different superscripts are significantly different (P<0.05).

0 100 200 300

Sample - 0.1 1 10 50 100 (µµg/mL)

a a a a a

b

0 10 20 30 40 50 60

SampleLPS - +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1 e

a ab b

c

d d

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1 e

a

b bc c

d d

0 500 1,000 1,500 2,000 2,500

c

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

d d

f

a b

e 0

100 200 300

Proliferation index Nitric oxide (µM) IL-6 (pg/mL) TNF-α (pg/mL) IL-1β (pg/mL)

0 20 40 60 80

b

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

a a

a a a

a

Fig. 5. Inhibitory effect of ethanol extract of Undaria pinnatifida root on the production of IL-1β in RAW 264.7 cells.

a-bMeans with different superscripts are significantly different (P<0.05).

0 100 200 300

Sample - 0.1 1 10 50 100 (µµg/mL)

a a a a a

b

0 10 20 30 40 50 60

SampleLPS - +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1 e

a ab b

c

d d

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

50

- +

0.1 e

a

b bc c

d d

0 500 1,000 1,500 2,000 2,500

c

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

d d

f

a b

e 0

100 200 300

Proliferation index Nitric oxide (µM) IL-6 (pg/mL) TNF-α (pg/mL) IL-1β (pg/mL)

0 20 40 60 80

b

Sample -LPS +

- + + +

1 10 + 100 (µµg/mL)

- + 50 0.1

a a

a a a

a

미역뿌리 에탄올 추출물의 항염증 활성

755

한

IL-1β

^와

TNF-α

^{는여러형태의실험에서상호상가효과를} 나타낸다는것이밝혀졌으며

,

급성기염증반응에서급격히증 가한다고알려져있다

(Lee, 2009). LPS

^{로염증반응을유도한}

RAW 264.7

세포에미역뿌리에탄올추출물을첨가하여

IL-1β

분비량을측정한결과

, LPS

^{처리구에서는}

62.60 pg/mL

^로증 가하였으나

,

^{미역뿌리에탄올추출물을농도별}

(0.1, 1, 10, 50, 100 μg/mL)

로처리했을때유의적인차이가없어큰효과를보 이지않았다

(Fig. 5).

따라서미역뿌리에탄올추출물은

LPS

에의한

RAW 264.7

세포의염증매개물질인

NO, IL-6

^및

TNF-α

^{분비를감소시켜} 항염증효과를가짐을확인하여천연항염증제로서이용가능 성이있는것으로확인되었다

.

사 사

위논문은

2014

^년도정부

(

^{해양수산부}

)

^{재원으로영남씨그랜} 트센터의지원을받아수행된연구임

.

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