Anti-inflammatory and Anti-tumor Effects of Tetragonia tetragonoides Extracts

번행초 추출물의 항염증 및 종양억제 효과

최혜정¹, 이성태², 권기석³, 주우홍¹*

1창원대학교생물학화학융합학부

2순천대학교약학대학

3안동대학교생약자원학과

Received: September 1, 2015 / Revised: December 4, 2015 / Accepted: December 4, 2015

염증반응은병원체와같은외부항원이체내로침입하거 나세포조직이손상받았을때활성화된면역세포에의해일 어나는생체방어를위한필수적인반응이다

.

cytokine, chemokine, NO

prostaglandin E2(PGE2)

^{[9, 14],}

^,

^,

질병이유발된다

^{[1, 5].}

조절할수있는물질은만성염증및자가면역질환에대한치 료제로사용될수있다

^.

한처방으로세포의

^DNA

^,

을억제함으로써효과를나타낸다

^.

^,

^,

고

^,

료를어렵게한다

^[7].

리학적연구는새로운약물개발을위한기초연구로서잠재 적으로가능성이있는소재를발굴하며나아가그기능을확 인함으로써신약개발에필수적으로중요한과정이다

^.

^,

^.

번행초는서양에서

“New Zealand spinach”

^,

^,

^{[8, 15].}

또한수용성다당성분은항염증활성이있는것으로보고되

어있으며

^{[8, 13],}

항균

^,

^{[2, 3,}

10].

환으로염증이유도된

^RAW264.7

분획물을처리하여항염증효능을검증하였으며

^{, Sarcoma-} Anti-inflammatory and Anti-tumor Effects of Tetragonia tetragonoides Extracts

Hye Jung Choi

, Sung-Tae Yee

, Gi-Seok Kwon

, and Woo Hong Joo

*

¹Department of Biology and Chemistry, Changwon National University, Changwon 641-773, Republic of Korea

2

College of Pharmacy, Sunchon National University, Sunchon 540-742, Republic of Korea

3

Department of Bioresource Science, Andong National University, Andong 760-749, Republic of Korea

We examined the anti-inflammatory effect and anti-tumor activity of Tetragonia tetragonioides crude extracts and fractions. The anti-inflammatory activity of T. tetragonioides was exuded through the inhibition of lipopolysaccharide (LPS, 1 µg/ml), induced nitric oxide (NO) and interleukin (IL)-1 β production. The production of IL-6 and tumor necrosis factor (TNF)-α also decreased in LPS induced RAW264.7 cells after treatment with polysaccharide (PS) fraction. Furthermore, the hexane (HX) fraction strongly inhibited the granulocytes macrophage-colony stimulating factor (GM-CSF) production. In ICR mice previously inoculated with Sarcoma 180, the life prolongation effects were 16.67% with an intraperitoneal injection of methanol (MeOH) extract and poly- saccharide fraction at a dose of 100 mg/kg/day. The results are an important preliminary step toward the development of effec- tive anti-inflammatory and anti-tumor agents using T. tetragonioides.

Keywords: Anti-inflammatory, anti-tumor, RAW264.7 cells, ICR mice, Tetragonia tetragonioides

*Corresponding author

Tel: +82-55-213-3453, Fax: +82-55-213-3459 E-mail: [email protected]

© 2015, The Korean Society for Microbiology and Biotechnology

180

^.

번행초건초

^{1 kg}

methanol(MeOH)

^1:3

⁶

³

^,

MeOH(70.0 g)

^.

hexane(HX, 14.35 g), dichloromethane (DCM, 8.97 g), ethylacetate(EA, 6.99 g),

^butanol

(BuOH, 5.15 g)

얻었다

^.

^MeOH

^{(58.0 g)}

^,

^DMSO

0.22

^m

⁴

^C

^.

실험동물은

⁽

⁾

⁽

^,

^,

⁾

서생후

⁶

^ICR

이트사육상자

⁽¹⁸

^{20 cm)}

⁵

²¹

²

^C,

⁵⁵

^10%

¹²

^.

(SCNU IACUC permit Number: 2013-1)

^.

RAW264.7

^,

^HL-60,

^A-549

Sarcoma-180

⁽

⁾

^.

sodium bicarbonate(NaHCO

, 2 g/l), 10,000 units/ml penicillin G sodium, 10,000 units/ml streptomycin sulfate, 25

g/ml amphotericin B, 50

M 2-mercaptoethanol(2- ME), 10% fetal bovine serum(FBS)

^{RPMI 1640}

. Nitric oxide(NO)

^Griess

^NO

. RAW264.7

⁵

¹⁰

³⁰

¹

^g/ml

^LPS

³⁷

^C,

5% CO

⁴⁸

^.

량의

^Greiss

¹⁰

^{, 550 nm}

^{, nitrite}

sodium nitrite

^.

또한 사이토카인 생성량 측정을 위해 동일한 조건에서

RAW264.7

^,

카인

interleukin(IL)-1

, IL-6, tumor necrosis factor(TNF)-

granulocytes macrophage-colony stimulating factor (GM-CSF)

(enzyme-linked immunosorbent assay, ELISA)

^.

^{, 1}

^goat anti-human cytokine

⁵⁰

^l/well

⁴

^C

overnight

^{, 10% FBS}

^PBS

^blocking

다

^.

^biotin

착된

²

¹⁰⁰

^l/well

응시킨 뒤

avidin-peroxidase 100

^l/well

2,2’'-azino-bis

^{405 nm}

^.

^cytokine

계치는

^{10 pg/ml}

^.

로

¹⁰

^{, MeOH}

군으로나누고이들마우스복강에

Sarcoma-180(3

¹⁰

⁾

^,

100 mg/kg/day

⁴⁰⁰

^l

PBS

^{. 3}

³⁰

^,

(median survival time, MST)

(increase in life span, % ILS)

^.

% ILS = (T/C

¹⁾

¹⁰⁰

[T:

^{MST, C:}

^MST]

번행초추출물및분획물의항염증효능측정을위해

^0.1

100

^g/ml

^,

¹⁰

^g/ml

. RAW264.7

^LPS

^NO

^{, 1}

^g/ml

^LPS

^NO

는

^MeOH

^,

히

^HX

(Fig. 1).

^inducible nitric oxide synthase

(iNOS)

^NO

화시켜조직손상및유전자변이및신경손상등을유발한 다

^[12].

⁴⁰

^g/

ml, 20

^g/ml

^NO

^40%

^{[6, 16],}

^HX

¹⁰

^g/ml

^60%

Fig. 1. Effects of solvent extracts or PS fraction on LPS-

induced NO production in RAW264.7 macrophages. Cells

were pretreated with the indicated concentrations of solvent

extracts or PS fraction for 30 min before 1 μg/ml LPS treatment

for 48 h. NO in culture medium was determined using the Griess

reagent.

자로서중요한작용을하는

^NO

^.

^IL-1

^, IL-6, TNF-

^GM-CSF

출물이

^LPS

^IL-6

였으며

^{, TNF-}

생성억제효능을보였다

^{(Fig. 2).}

^HX

포증식을조절하는

^GM-CSF

과적으로작용하였으며

^{, TNF-}

^DCM

^.

^.

^HX

-iso-cubebene

^{, lignans}

활성성분의함량도높은것으로보고되고있는바

^[4],

구에서도

^HX

^.

편번행초의수용성다당류분획물을

^{25 mg/kg}

스에처리했을때염증반응의하나인부종을약

^35%

^[8],

^{(1, 6)}

^3-O- actyl-

-glucose

^-

-arabinofuranose

^-

- rhamnopyranose

^.

cerebrosides

steryl glucoside

가있는것으로보고되고있다

^[15].

부종억제효과를보이는항염증활성를입증하기위하여구 체적인연구를진행하였으며이러한결과는번행초의

^HX

^,

^.

한편예로부터위암등항암효과가있는것으로알려진 번행초의항암효과를조사하기위해혈액암세포주

^,

^,

(unpublished data).

효능은마우스를

³

Sarcoma-180

주사하여복수암을유발시킨뒤

^,

^{(Fig. 3).}

^,

¹⁸

²⁰

으로나타났다

^{. MeOH}

속도가느렸고평균수명은가장긴

²³

^,

^. 18

²⁵

^,

^21.7

^.

Table 1

^.

^MeOH

지속적인

^DNA

Fig. 2. Effects of solvent extracts or PS fraction on LPS-induced production IL-1 β, IL-6, TNF-α and GM-CSF in RAW264.7 mac-

rophages. Cells were pretreated with the indicated concentrations of solvent extracts or PS fraction for 30 min before 1 μg/ml LPS treat-

ment for 24 h. The amounts of IL-1 β, IL-6, TNF-α and GM-CSF were next determined using the ELISA test.

추는기작으로수명을연장시키는것으로사료된다

^.

^,

써부작용이거의없는것으로알려져있다

^[11].

당성분은

in vitro

보고되어있으나

^,

^.

최근연구개발되는항암제는대부분복약순응도를높이 기위해경구제로투여되고있는추세이므로

^,

in vivo

^,

특히번행초는오래전부터식용으로사용되고있으므로경 구제제로도개발가능하다

^.

^.

요 약

본연구에서는번행초의추출물과분획물의항염증효과 및항암활성에대해연구하였다

^.

1

^{g/ml LPS}

^NO

^IL-1

통해확인되었다

^{. IL-6}

^TNF-

¹

^{g/ml LPS}

RAW264.7

소되었다

^.

^HX

^GM-CSF

게억제하는것으로확인되었다

^.

Sarcoma 180

^ICR

^MeOH

100 mg/kg/

day

^16.67%

^.

^.

Acknowledgments

This research is financially supported by Changwon National University in 2015-2016.

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Median survival (days)

Life span (%)

Increase of life span (%)

Control 18 100 -

MeOH

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PS

21 116.67 16.67

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MeOH extract

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Polysaccharide fraction

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