Anticancer Effect of Erythronium japonicum Extract on ICR Mouse and L1210 Cells with Alteration of Antioxidant Enzyme Activities

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(1)KOREAN J. FOOD SCI. TECHNOL. Vol. 36, No. 6, pp. 968~973 (2004). ©The Korean Society of Food Science and Technology. â.æ ºÂb~ ICR îÖÊf L1210 z^

(2) ö & z·Ï ö 8 ÖzÎ² Wæz .êÁ;'1Á~*ãÁ;ö* ç«L z, 1] ~ \ö £*Ò **¦. Anticancer Effect of Erythronium japonicum Extract on ICR Mouse and L1210 Cells with Alteration of Antioxidant Enzyme Activities Yoo Jin Shin, Dae Young Jung1, Hye Kyung Ha, and Sie Won Park* Department of Chemistry, Sangmyung University Department of Herbal Pharmaceutical Medicine, Korea Institute of Oriental Medicine. 1. Effects of Erythronium japonicum methanol extract on ICR mouse with induced abdominal cancer and L1210 cells were studied. Administration of methanol extract (10-100 µg/20 g body weight) prolonged life by 47.8% and decreased number of L1210 cells with IC50 of 54.6 µg/mL after 3 days culture, whereas little effect was observed against normal lymphocytes (<6% compared to 83.2% of L1210 cells under the same condition). Increased SOD and GPx enzyme activities, and remarkably augmented generation of O2− ion in L1210 cells by E. japonicum extract, implied that reactive oxygen species including O2− ion, might have participated in L1210 cell death Key words: Erythronium japonicum, anticancer activity, ICR mouse, L1210 cells, superoxide dismutase, glutathione peroxidase. >Ú oncogenes Wz>¾ suppressor genes ÛB>B ; æ² >Ú Vº ©b " > ®º ç ç~ /B¶ Úö F ~ã¾ bîö ~ W >º WÖ²(reactive oxygen species, ROS)& >Ú^ z. (6,7). Ú ©f ªç'b ÒÏ> ®º zB doxorubicin, cisplatin, vincristine, cytosine arabinoside, methotrex, bleomycin ~ ·ÏV·(8-11) WÖ²f ç7 * 7b &~ apoptosis W >Ú z^¢ ÒÊ º ©b ¾æ¾ Ö WÖ²º Bz~ öª" ÿö z~ > B ~º 6 . öBº ÿn ZëW ¾b¾ *ªöò ÒÏ> ÚN â.æ~ zêRºÂbö &~ Sarcoma 180b > zj FBÎ ICR îÖÊ 5 W.÷ê z^" L1210 ^ ö & zÎ"¢ þ~ ¾j& normal lymphocyteö & ëW~ ;ê¢ ¦ï~, ·ÏV·j «~¶ W Ö² O2−N~ ;ï 5 ÖzÎ² SOD, GPX, catalase Î²Wj G;~ ~ ®º Ö"¢ áÚ B~º : .. â.æ(Erythronium japonicum Decne)º W"(Liliaceae)ö ³~º j .bB N*#Ö¶, ÖÖv, &BZ, j ¾b¢ ®ÒÖVê ~, . 2¦V 5úræ pf Ö öB zJ~ jª Ú Ej bÖº bB ¸& 25-30 cm >~ ãf k³ p 30 cm ræ Ú&, ÚÖ ´f " ¾b ÒÏ> ãöBº öj j~ ®b ÒÏ z . â.æ FÒ b~ ãö FB ö ïf >W g/kgræ (1), ö 7öº ;º*¢ ÎÚº osmoprotectant 5 z·Ïê > ®º fructan (2,3) çï >Ú ® . *Ò zBº Vzj j z~ò~ *";ö &¦ª R> ®º ©f zB& *'b ·Ï~V r^ ö ~ò 5 *¢ .O~V *~ F^> ® " > ®. . ¾ zB& z^ö & zÎ" ;ç^ö &BN ; ëW" BÚW(4,5)" ?f ¦·Ïj ¾æ ÚV r^ö zB~ ÒÏf ~¶~ ~òö ÂB& > ®. . V¢B ' f &Ë~ ZëW¾ ëW Ö £~ Bê z·Ï Ö> zB ö .j VÞ ® b¾ jçræê ç' zBBBf Úææ p ® . Þ z¦ genetic changesf epigenetic changes& ö. þÒò 5 O» £ Ferricytochrome c, xanthine, xanthine oxidase, superoxide dismutase, glutathione, glutathione reductase, phorbol 12myistate 13-acetate, nicotinamide adenine dinucleotide reduced, trypan blue, dimethylsulfoxide, ethylenetetraacetatef Sigma(St. *Corresponding author: Sie Won Park, Department of Chemistry, Sangmyung University, Hongji-dong, Seoul 110-743, Korea Tel: 82-2-2287-5147 Fax: 82-2-396-8758 E-mail: [email protected] 968.

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(4) jÞ r ¾² Ú r jN ºÂbj B~& . ò 10 gj 100 mL~ 70% zêRö æ~ 24* N O~ ê çû j ~ ºÒ¢ ÿ¢~² ¾Ò~ çûj . B áf çû" ~ 7ß~ gauze¢ Û"* áf j Watman "æ "* j 6{ ³». r ïÿ ~ Z²¢ ïï~& . Bz ICR îÖÊ~ « ËÎ ICR îÖÊöB ê&V· >Ú N Sarcoma 180 cellsj þ Ï ICR îÖÊ ;ö 1Ü105 cells/10 µL/20 g body weight~ ³ê 7« r 24* ê¦V, â.æ ºÂbj 0.5% Ú~ ²~ DMSOö Ï~ 10-100 g/20 g body weight~ ³ê ¢ ¢; *ö Nf O ãR r ç¢ &V~B Nj êÖ~& . Control~ ãÖº òö Ò Ï ÿ¢ Ïï~ DMSO¢ Ã~>ö &~ ÿ¢ O»b ã R ~& . L1210 ^

(5) " V· öB ÒÏ L1210 cells(murine lymphocytic leukemia; ATCC CCL 219) ^º ^" f¯(BÞ& z bV)öB ª· Aj ÒÏ~& . z^º 37oC, 5% CO2, humidified conditionöB V·~&b V·f 10% FBS¢ F RPMI 1640î, Bº penicillin/streptomycin~ b j ÒÏ~&b ê& V·f 3-4¢î ¯~& . Normal lymphocytes ªÒ ;ç^ö & ZëW~ ;ê¢ ¦ï~V * ICR îÖ Ê¦V normal lymphocytes¢ ªÒ~ ^ö ~º â .æ ºÂb~ ëWj ¦ï~& . Normal lymphocytes~ ªÒ O»f Boyum(12)~ O»j 'Ï~&º *® r" ? . Mouse¢ ether îÎ ê ~&; j .B~ heparin ¾ Ò& B "ÒVö j. Ê 15 mL tubeö ÎjB Vö dextran FêÚ Histopaque R-1077 3 mLj Ê#² &. r 400ÜgöB 30ª* NöB öªÒ ~& . öªÒ ê pasteur pipette çû plasma[j B~ lymphocyte & B ®R« 7*[j ~ phosphated buffered saline (PBS) 10 mL" Ê#² b r 250ÜgöB 10ª* ö ªÒ ~ çûf ªÒ áÚê *bö & 2² > ^¿~& . áÚê normal lymphocytes¢ PBSö ¦F* ^ >¢ êÖ r penicillin/streptomycinÏ" 10% FBS& FB RPMI 1640 Væö &~ V·~& . L1210 ^

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(7) > 6² â.æ ºÂb ¾æÚº L1210 z^ Ã ÛBÎ"f normal lymphocytesö & ëW;ê¢ ^ > æz¢ æ. 969. ¯~& . 24 well plateö 1Ü106 cells/mL ³ê L1210 ^ 6º normal lymphocytes¢ &~ Vö â.æ ºÂbj Î& r ¢;V* V· ê Thayer(13) O»ö &~ hemocytometer¢ ÒÏ trypan blue exclusion»b Új®º jOï ^>f control ~ jOï ^>¢ ^Ú WªN (%) ^ëWj êÖ~& . ¯ control ~ > 6² >¢ ^ëW~ Ö"~ ¢~b *"~ êÖ~& . ^> 6²(%) = Control~ jOï ^>−ò~ jOï ^> Ü100 Control~ jOï ^> L1210 ^

(8) ~ O2− N ;ï L1210 ^ V·ö â.æ ºÂbj Î&~ ¢;V* V· ê L1210 ^& ¾æÚº O2− Wï~ ;ïf Markesbery(14)~ O»ö ~~ ¯~& . þöBº L1210 ^ ¢ 1¢-3¢* V·~ 250ÜgöB öªÒ ê çûf ª Ò >÷~ 24 well plateö 1Ü106 cells/well, 700 µL PBS, 50 µL cytochrome c, 5 µg/mL~ phorphol myristate acetate (PMA) 50 µLf Væ¢ Î&~ 60ª* V· ê öªÒ ~ & . çû 200 µLö PBS &~ 1 mL& >ê C~ 550 nmöB 7ê¢ G;~& . Ferricytochrome c~ ~ö >wf 21.1/mM/cm~ Ö7ê>¢ 'Ï~&b O2−Wïf moles/60 min/108 cells ¾æÚî . ÖzÎ² B L1210 ^¢ 1Ü106 cells/mL ³ê 7« V· 10 mL ¢ 50 mL culture flasköB ¢;V* V· ê 1,000ÜgöB öªÒ ~ çûf B~ *B L1210 ^¢ Ò "> 2² ^¿~& . ÷B ^¢ 5 mM sodium phosphate buffer(pH 7.4) 1.5 mLö *ç~ Elvejehn homogenizer ¢ Ï~ ice bathÚöB 5.O 5² îz r îö &~ .r2 2êV¢ Ï~ âr çöB 30.* 3² .r2 ¾Ò~ mitochondria matrixö Ò~º ©b r Jê MnSOD Î²ræ ÏÂ* îj 4,500ÜgöB ö ªÒ~ çûj ÖzÎ²b B~& . Superoxide dismutase(SOD) W /; SODW~ G;f V'b McCord and Fridovich(15)~ O»ö ~ Úr . 3 mLö 50 mM potassium phosphate buffer(pH 7.4), 100 mM cytochrome c, 50 mM xanthine, 0.1 mM EDTA(pH 7.8), Î² B Ïj 25oCöB 15ª* .~ r xanthine oxidase(XOD)¢ Î&~ >wj B~ &b >wf 550 nmöB 10. * 5ª* 7ê¢ G; ~ Úr . Xanthine oxidase Î&ïf Î²j F~ æ pf >w~ 7ê >& ª ² 0.025& >ê .~& . SOD Wf Î² F>æ pf >w~ cytochrome c~ ~ö³ê¢ 50% ÛB~º ·j 1 unit k; ~ unit/min/107 cells ¾æÚî . Glutathione Peroxidase(GPx) W /; GPx~ Wf Maral (16)~ O»ö ~~ G;~& . 50 mM potassium phophate buffer(5 mM EDTA, pH 7.0), 8.4 mM NADPH, 1 unit glutathione reductase(GSSGR), 150 mM~ glutathionc(GSH), Î²j Î&~ *Ú >w 2 mL >ê.

(9) ®"²æ B 36 ² B 6 ^ (2004). 970. r 37oCöB 5ª* .~Î ê 0.16 mM t-butyl hydroperoxide¢ &~ 340 nmöB 3ª ÿn NADPH~ Öz ö ~ 6²>º 7ê~ æz¢ G;~&b NADPH~ 7ê>º 6.22 cm−1mM−1j ÒÏ~ Î²Wj êÖ~& . CatalaseW /; Catalase W~ G;f 250 nm, 25oCöB 7ê 6²¢ " ~º Maral (16)~ O»j 'Ï~& . 50 mM potassium phosphate buffer(pH 7.4), 12.5 mM H2O2 5 Î²j &~ *Ú >w 2 mL& >ê ê 250 nmöB 10. * 5ª* 7ê æz¢ G;~& . Cê¾Ò. þ Ö"º meanÛS.D.b ~& . Control8" ò 8 Ò~ F~Wf Student’s t-test¢ 'Ï~ p* < 0.05, p** < 0.01 ãÖ¢ F~W ®º ©b 6;~& .. Ö 5 V â.æ ºÂbö ~ Bz ICR îÖÊ~ «ËÎ Sarcoma 180b BzÎ ICR îÖÊö â.æ ºÂbj ã R~ ~º V*j control~ ©" jv~ êÖ «ËÎ"º Table 1ö :f ? . ICR îÖÊö >zj FB~V *~ Sarcoma 180 cellj 7«~ 24 * æÂ ê¦V â.æ ºÂbj 10-200 µg/mL ¢ N fO ãR ~ Ò¢j êÖ Ö" control~ ï V*f 21.3Û3.7 daysîb â.æ ºÂb R~ «ËÎ"º 50 µg/mL~ ³ê¦V *&® Ã&~ 70 µg/mL ³êöBº 143.7% ö ~&b *Ú'bº Table 1. The life prolonging effect of the Erythronium japonicum extract on ICR mice bearing peritoneal cancer Erythronium japonicum (µg/20 g body weight). Survived days. Life prolonging effect (%). 00.0 10.0 20.0 50.0 70.0 100.00. 21.3Û3.7 24.6Û4.3 23.8Û3.9 *28.7Û2.2* *30.6Û1.6* 29.5Û4.7. 1000. 115.5 111.7 134.7 143.7 138.1. Each value represents the meanÛS.D. of six individual experiments. Asterics denote a significant difference compared with the control group (*p<0.05).. 111.7%-143.7%ö ~& . â.æ ºÂbf >zj FBÎ ICR îÖÊ~ «j ç® ËÊº ©b 6 >î . â.æ ºÂb~ L1210^

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(11) ëW â.æ ºÂb zFB ÿb&*öB ç «ËÎ" ¢ ¾æÚîbæ z^" &*öB~ ÛBÎ"¢ ¦ï ~ . W.÷ê z^ L1210 ^ö â.æ ºÂbj 10 µg/ mLöB 200 µg/mL ³êræ Î& r 1-4¢* V·~ ^ >¢ êÖ Ö"& Table 2ö B>Ú ®º V· 1¢ öº ^> 6²& ~ ¾æ¾æ p~b¾ ³ê& Ã&~ V·¢>& Ã&> *& ^> 6²¢ ¾æÚî . f ?f ^> 6²º ^ëWb ~ z^¢ Ò Ê¾ 6º cell cycle arrestf ?f ^ª ÛB r^ © b ÚÆ FöB z^>& 6² º ©f zÎ "~ 7º æ . V· 3¢" V· 4¢öº 50 µg/mL ³ê çöBº £ 50%ç~ ^ëW Î"¢ ¾æÚî, 4¢* V· öº 98.6%~ ^ëW Ò~º :f ? ³êöBº ÚjÎf ^& ~ ìî . ^ëWö & ¢>' þ V& V· V* 3¢j F~ êÖ IC508f 54.6 µg/mLî . Normal lymphocytesö & â.æ ºÂb~ ëW â.æ ºÂb zBB~ Î"& ®bJ z^ö & Bº ^ëW ¸f & ;ç^ö &Bº ZëW {>Ú¢ ~æ L1210^& F¾ ICR îÖÊ¦V normal lymphocytes¢ ªÒ~ ëWFZ¢ þ~& . Table 3ö BB Ö"ö V normal lymphocytesö â.æ ºÂ bj Î& ê V· 1¢öº &¦ª ^Ò º 10% Ú~ ^ÃÎ"& ¾æÒb, V· 2¢öê & ³ê~ ãÖöº ^Ã Î" Ò 100-200 µg/mL~ ³êöB 5% Îù~ £ ^> 6² Î"& ¾æÒ . V· 3¢ öê £ 5% ;ê~ 'f ^> 6² *çî V· 4¢ ö &B¢ 10%& >º ^ëW Î"& ¾æÒ . Ö"¢ L1210 ^~ ãÖ Table 2~ Ö"f jv~ â.æ ºÂbf z^ö &Bº ¸f ^ëWj ¾æÞ > normal lymphocyteö &Bº j" £ ëWòj ¾ æÚî . .¢ Ú â.æ ºÂbj 100 µg/mL³ê &~ 3¢* V·8j r L1210 ^~ Nf 16.8%;êö ® "~&b¾ normal lymphocytesº 84.2%~ ¸f Nj &. . V¢B â.æ ºÂbf BB~Vö V¢Bº ëW ' zWf ¸f Ö> zBB~ &ËW ÒB " > ® .. Table 2. The effect of Erythronium japonicum extract on the cell number changes of L1210 cells Cell number reduction (%) Culture days 1 2 3 4. Extract concentration (µg/mL) 10. 20. 50. 100. 200. 1.7Û0.5 2.1Û1.1 15.5Û2.30 16.7Û3.70. 1.4Û0.8 5.7Û3.8 23.8Û5.70 32.5Û3.9*. 3.0Û1.9 26.0Û1.3* 53.0Û2.1* 072.4Û3.6**. 5.3Û1.60 39.2Û5.1** 83.2Û2.5** 83.7Û3.4**. 7.4Û1.2 46.2Û4.9* 095.7Û3.3** 098.6Û2.3**. Each value represents the meanÛS.D. of four individual experiments. Asterics denote a significant difference compared with the control group (*p<0.05, **p<0.01)..

(12) â.æ ºÂb~ ICR îÖÊf L1210 z^ö & z·Ï" ö V ÖzÎ² Wæz. 971. Table 3. The inhibitory effect of Erythronium japonicum extract on the cell numbers of normal lymphocytes Cell number reduction (%) Culture days. Extract concentration (µg/mL). 1 2 3 4. 10. 20. 50. 100. 200. -9.2Û1.3* -2.5Û1.20 1.4Û1.5. 2.5Û0.3.. -4.6Û2.2 -2.1Û0.7 0-3.2Û0.5* -5.3Û1.4. -7.1Û3.4. 1.5Û0.4 4.2Û1.3 005.9Û0.6**. -4.3Û0.6*. 3.4Û0.80 05.8Û0.4** 15.9Û1.3*0. 3.8Û1.7 5.9Û1.7 6.7Û1.5 17.3Û3.50. Each value represents the meanÛS.D. of four individual experiments. Asterics denote a significant difference compared with the control group (*p<0.05, **p<0.01).. L1210 ^

(13) ~ O2− W â.æ ºÂb z^ L1210 ^>¢ *&~² 6² * ¸f zÎ"~ &ËWj &º f ?f z^> 6 ²Î"& ÚÆ V·ö ~ ¢Ú¾ºæ «~¶ z^ > 6² *ç" WÖ²f~ &ê¢ «~º þj ¯~ & . *Ò B zB~ ·Ï V·f zBö ~ z^ Ò necrosis º apoptosisö ~~ apoptosis /B ¶B~ WÖ²~(8-10)~ 'Ëf Ö æ â.æ ºÂ bö ~ z^> 6²öê f ?f WÖ² &êbî &>ºæ ¦ï~¶ ~& . ÖF â.æ ºÂbö ~ z ^> 6²& *&~² FBF > ®º j J;~ ö V O2−N~ Wï~ æz¢ G;~, 8j ^ë W FB>æ pf öB~ O2−N~ Wï" jv~ ~ . O2−Nf Ö²¦V W>Ú 2>º WÖ²& Òb~ ÑÂBbî Wïf ~& > ®Æ. (11,12). ^> 6²*çf â.æ ºÂb~ ³êf V· V *ö ~~æ â.æ ºÂbj 50 µg/mL" 100 µg/mL~ ³ê Î&~ 3¢* V·j Î ãÖ¢ ^ëW ¸f ãÖ *"~ 10 µg/mL" 20 µg/mL~ jv' 'f ³ê 1¢*¾ 2¢* V· ãÖ¢ ^> 6²& ~ ¾æ¾ æ pf ãÖ þ~& . f ?f öB '' V· ê ÷ ^ö & O2− Wïj G;~ jv Ö" Table 4öBf ? ç® F~W ®º O2− Wï~ æz¢ á² >î . Ö"ö V Table 2öBf ? ¯Vöº ^> 6² & 10% ~ Ö £ V· 1¢¦V â.æ ºÂbj Î& ~ O2− W control ö j~ 6ê'b Ã&~V ·~&b, ^> 6²& 50-80% ç® ¸ Table 4. The effect of Erythronium japonicum extract on the generation of O2− in L1210 cells O2− (nmoles/60 min/107 cells) Concentration (µg/mL) 00.0 10.0 20.0 50.0 100.00. Culture period (days) 1. 2. 3. 0.09Û0.05 0.13Û0.08 0.26Û0.04 00.34Û0.07* 0.45Û0.13. 0.14Û0.03 0.18Û0.06 00.67Û0.05* 1.32Û0.13 01.79Û0.17*. 0.11Û0.05 00.38Û0.07* 0.93Û0.11 01.47Û0.05* 02.35Û0.13*. Each value represents the meanÛS.D. of five individual experiments. Asterics denote a significant difference compared with the control group (*p<0.05, **p<0.01).. ~~ V· 3¢öº ³êö jf~ O2− W /Ã~&º 50 µg/mL~ ãÖöº control 8~ £ 12V, 100 µg/mL~ ³êöBº control 8~ £ 23V ;ê~ Ö ¸f Ã&¢ & . Ö"¢ «~ L1210 ^ö â.æ ºÂbj Î & ãÖ ¯bº ^> 6²& ¾æ¾æ pº V· ÑÆ ¦V O2−N~ W Ã&>V ·~ ^> 6² ~ ;êf jf~ æ³'b Ã&~º ©j " > ®î . ¢>'b O2−N~ >6Vº &Û 10−5 secb Ö ³~ ² &Ò(14)>º ©b rJ^ ®º, &Ò~ Ñ ® ê º O2−& H2O2f OH *~>º >w, v ® êº WB H2O2& ZëW H2O *~>º >w(11,12) . "; 7ö Vº H2O2º b O2−f ¦Öb ÁOHê. Ö F~ z^ö²ê Ú bî æ z^ WÖ² ö & ¶j ^~V * &w >wj ©b *">Ú ¦ïV ~& . ¯ â.æ ºÂbj &~ L1210 ^ Úö Z O2− Z ë H2Oræ *~>º ê>w~ æbî ÖzÎ² (17,18) superoxide dismutase(SOD), glutathione peroxidase (GPx) 6º catalase Wj G;~ â.æ ºÂb~ ^> 6²Î"f ê~ ~ . â.æ ºÂbö ~ L1210^

(14) ~ SOD W æz O2−f SODö ~B H2O2 *~> H2O2º catalase¾ GPxö ~B Zë H2O *~B rJ^ ®¾ Fenton >wö ~ ¶B'b ëW ÁOH *~>º ©b r J^ ®º(11,12), â.æ ºÂb ¢V>º ^ëW "; ö B SODW æz& >>>ºæ 2k~V *~ ^> 6²ö V SODWj '' G; jv~& . Fig. 1ö BB Ö"¢ â.æ ºÂbj Î&~ ¢ VB ^> 6² ;êö jf~ SODW~ æz& © j r > ®î . ÖF ^> 6²& ~ ¢Ú¾æ pº V· 1¢ö â.æ ºÂbj Î&~ V·Î ê L1210 ^~ SODWf 10 µg/mLöBº 1.31 units/107 cells, 50 µg/mL~ ³êöB 0.96 units/107 cells, 100 µg/mLöBº 1.64 unita/107 cellsB control 8 0.57 units/107 cellsö j~ £ 2-3V Ã& 8j & . f ?f *çf ¯bº ^> 6 ²& *&~æ pf V· 1¢öê B 'bº z^& â.æ ºÂbö ~ ëW ç& >Ú O2−¢ WÊ O2−¢ B~V * SOD Wê ²¦V Ã&~V · ©b . ^> 6²& 50-90% ;êræ ¸~~ V· 2¢f 3¢, 4¢~ Ö"¢ control ~ SOD Wê V· V*ö jf~ 2-3V Ã&~&b, â.æ ºÂb Î&~ SOD.

(15) 972. ®"²æ B 36 ² B 6 ^ (2004). Fig. 1. The effect of Erythronium japonicum extract on the SOD activities of L1210 cells. Data represent the mean ÛS.D. of five individual experiments. Asterics denote a significant difference compared with the control group (*p<0.05, **p<0.01).. 3V;ê ræ ² Ã&~& . WÖ² *~Î² SODf GPx ÖzÎ²ö & ç ~ Ö"¢ «~ L1210 ^ö â.æ ºÂbj Î& ã Ö *&~² ^> 6² *ç FB>º r O2−N~ W /Ã~, O2−Nj ²~V * SODW Ã &~, Ö" H2O2& Ã&~ H2O2¢ ²~V * GPx Î²~ W Ú ² Ã&~º ©b 6>î . GPxº " ^îö 6Ò ª>Ú ®º >, H2O2 ¢ ª~º 6 Î² catalaseº ^Ú peroxisomeö Ò>Ú ®b(18,19) GPx ªC þöBê ¦&~ catalase Î²~ Wj G;: Ú¶ öBê W ¾æ¾æ p ~º º catalase& peroxisomeö Ò>Ú ®ÚB >w B'¾ 6º WB H2O2 ªöº &~æ p, ^ > 6²ö >>>Ú &¦ª~ H2O2º ^îö 9² Ò~ º GPxö ~ ª >º &ËW ©b 6>î . ç~ Ö"¢ «~ â.æ ºÂbf L1210 ^¢ Â ]~² ÒÊ normal lymphocyteº ~ 2Z~æ pb, L1210 ^>¢ 6²Êº "; 7ö ÖF L1210 ^Úö O2−j ·Ö~ ¦V WB H2O2 6º ÁOH ~ W Ö²& apoptosis¢ FBb z^& ÒF &ËW Ò>îb, Þb /ÃB WÖ²~ ëWö &~ z^º WÖ² ²Î² SOD¾ GPx¢ FB~ W Ö²~ ëWj ïjJ ~¾ Öf ¦b Ò~² > º ©b *">î .. º Fig. 2. Effect of Erythronium japonicum extract on the GPx activities of L1210 cells. Data represent the mean ÛS.D. of five individual experiments. Asterics denote a significant difference compared with the control group (*p<0.05, **p<0.01).. Wf ''~ control 8~ 5V ;êræ~ ¸f Ã& 8j ©b j â.æ ºÂb ~ ¢V>º ^> 6 ²öº O2− *~Î² SOD Î²W ² >>>º ©j r > ®î . L1210 ^

(16) ~ GPx(glutathion peroxidase)f catalase W æz ç~ Ö"öB SODW Ã& º ©f Wb H2O2 Z º ©j ~~æ Ú H2O2~ *zÎ² glutathione peroxidase(GPx)¾ catalaseW~ æz& >>F >ê ® jê . Fig. 2~ Ö"ö ~~ SODf ÿ¢ öB GPx Wæz¢ G; : SOD~ ãÖf îR&æ ^> 6²*ç Â] öB GPx Î²Wf ^> 6² ;êö jf~ Â]~² Ã&~º ©j r > ®î . ÖF ^> 6²& ~ ¾æ¾æ pº V· 1¢ö â. æ ºÂbj 10 µg/mL¦V 100 µg/mL ³êræ Î&~&j ã Ö GPx Wf 0.7-1.2 unit/107 cellB control~ 8" ~ FÒ~&b¾ ^> 6²& *&®~ V· 2¢, 3¢ Ò 4¢~ GPxWf control ö ®ÚBê £ 2V ;ê, Ò ''~ â.æ ºÂb Î& ö ®ÚBê V· Vf ³êö j.~ GPx W B V·V*~ control 8~. £. ¾b¾ *ªöò ÒÏ>Ú N â.æ(Erythronium japonicum)~ z·Ïj ¦ï~& . â.æ ºÂbj Sarcoma 180 b >zj FBÎ ICR îÖÊö &~ ãR Ö " 143.7%ö º «ËÎ"¢ áîb, W.÷ê z^ L1210 ^ö &Bº 98.6%~ ^> 6²Î" ¢ áî . jÞ â.æ ºÂb~ ëWFZ¢ ¦ï~¶ normal lymphocyte¢ ªÒ~ ;ç^ö â.æ ºÂbj Î&®j r ^> 6²º ~ ¢Ú¾æ p~b, ³ê~ 3¢*~ Z V·V*ö ~Bê L1210 ^~ ãÖ 83.2%ö j 5.8% ;ê Ö 'î . f ?f Ö"¦V â.æ ºÂbf ZëW 6º &ëW zBB~ BB &~& ®j ©b Òò>î . Þ â.æ ºÂb~ z^ö & ·Ï V·b ¢~b WÖ² O2−~ Wïj G; Ö" control 8~ & £ 3.6Vræ ² Ã&~&b, ÿö O2− *~Î² SODf SOD~ «Öb H2O2 ªÎ² GPxê '' 5Vf 3Vræ ² Ã&~& V¢B â.æ ºÂbf z^öB O2−¢ j WÖ²¢ FBÊ Ã&B WÖ²& z^~ apoptosis¢ ¢V~º ©b *">î .. ^. ò. 1. Mullin WJ, Peacock S, Loewen DC, Turner NJ. Macronutrients content of yellow glacierlily and balsamroot; root vegetables used by indigenous peoples of north western north America. Food Res. Intl. 30: 769-775 (1997) 2. Ritsena T, Smeekens M. Fructans: beneficial for plants and humans. Curr. Opin. Plant Biol. 6: 223-229 (2003) 3. Cooper PD, Carter M. The anti-melanoma activity of inulin in mice. Mol. Immunol. 23: 903-911 (1986) 4. Astrow AB. Rethinking cancer. Lancet 343: 494-503 (1994).

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