Antioxidant Effect of Rosa davurica Pall Extract on Oxidation of Human Low Density Lipoprotein (LDL)

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(1)KOREAN J. FOOD SCI. TECHNOL. Vol. 36, No. 2, pp. 311~316 (2004). ©The Korean Society of Food Science and Technology. Ò²~ &ê æOWî~ Özö & æ¾Z ºÂb~ Öz Î" ÒÒî*Á jÁÆÁ;ãêÁ

(2) ÁJ+ÁB'êÁ;~^ÁB%81ÁR91 ;öê~ã\ö, 1;.`³ë8F>8. Antioxidant Effect of Rosa davurica Pall Extract on Oxidation of Human Low Density Lipoprotein (LDL) Jae-Hoon Sa*, Woan Lee, In-Cheol Shin, Kyung-Jin Jeong, Tae-Heum Shim, Heung-Seok Oh, Yong-Jin Kim, Eui-Ho Cheung, Gwang Gee Kim1, and Dae-Sung Choi1 Gangwon Research Institute of Health and Environment 1 Jungsun Agriculture Technology Center Antioxidant effects of Rosa davurica Pall extract on copper-mediated LDL oxidative modification were investigated. Oxidation products of LDL were determined based on TBA value, formation of conjugate diene, and apolipoprotein carbonyl value. As revealed through TBA values, ethyl acetate and butanol fractions of R. davurica Pall root showed strong antioxidant effect, with 85.3 and 93.2% inhibitions at 30 µg/ml each, respectively. Ethyl acetate and butanol fractions at 30 µg/mL inhibited LDL oxidation up to 8 hr. Conjugate diene formation by lipid oxidation with Cu2+ addition in ethyl acetate and butanol fractions decreased 2.2- and 5.6-fold, respectively, compared to control. Carbonyl value decreased in the presence of butanol and ethyl acetate fractions. Methanol and ethyl acetate extracts of R. davurica Pall root showed higher absorbancy at 285 nm. Ethanol extract of R. davurica Pall root and stem contained 10.6 g/100 g total phenolic compounds. Results reveal phenolic compound as major biological component in R. davurica Pall extracts. Ethyl acetate and butanol fraction showed strongest antioxidant effect on LDL oxidation. Key words: low-density lipoprotein(LDL), antioxidants, Rosa davurica Pall, phenolic compounds. *. . & . apo B-100f LDL-receptorö Ö> ®º ligand ·Ï . ;ç' LDLf oxygen free radicalö ~ £² Öz>Ú LDL>ÏÚf Ö~æ á . LDLÖzº . V ÿ ãzW ÷æ~ ;W" ê*ö "º j ~, Ö zB LDL ;ç' LDL &^ö ~ £² ³> Ú ®^(foam cell)¢ ;W~ ÿ ãz¢ FB (2-5). f? æî~ "Öz Oæf LDL~ Öz¢ Oæ~V * ~, BHTf BHA ~ WÖzB¢ ô ÒÏ~ ®b ¾, zj FB º & ®b, Úö ëWj ¾ æÚÚ ÒÏB¢ A ® . >ö ÂÖzB Ï> ®º α-tocopherol 5 jæ"-Cf Öz Î"& Ô &Ï ç&'b j" Wî ® . ö β-carotene, αcarotene, lycopene, retinodes, 5 ubiquinol ®º, f &¦ª bö Ò~º ©b rJ^ ®b polyphenol, flavonoid 5 FêÚ& ; Öz ·Ï ®º ©b r J^ ® (6-11). æ ÖzË Ö>~ Úö Z ÂÖzB¢ b¶öb¦V BB~Jº æçª B~. . æ.ræ B ÖzBö & º f¯´ ºÂb ~ LDLö & ÖzÎ"(12), ºÂb~ ÖzÎ"(13), ."(14), N~ Öz W(15), " .(16) ® .. " >&~ Ëçö ~ æOî~ " Sº Ë 5 .&ê î~~ /Ï Ã&¢ &^J² >î, ¢ V>º æ.Ãf &&êæOWî(low density lipoprotein, LDL)~ ç&Ò ¢Ú¾º #çæzf &N ¸, ß® & çÿ Ã& Ëî~(coronary disease)~ Bö 7º *þ¶ ÖÒ¾¢ W÷ Ò~ >*¢ Næ~ ® (1). LDLf plasma protein7 density 1.025öB 1.058ö ~º lipoproteinb hydrophobic triglyceride cholesteryl ester Ú^ ®b, 7¦º W phospholipidf free cholesterol ~ ® . ¦º apo B-100¢ ®Òº ß>Wî W>Ú ® . Apo B-100f LDLöB monomeric proteinb Ò~ ÚÚ .&öB cholesterol~ . 5 &Òö ç7. *Corresponding author : Jae-Hoon Sa, Gangwon Research Institute of Health and Environment, Chuncheon, Gangwon 200-822, Korea Tel: 82-33-645-0830 Fax: 82-33-645-0839 E-mail: [email protected] 311.

(3) ®"²æ B 36 ² B 2 ^ (2004). 312. æ¾Z(Rosa davurica Pall)º Ë"ö ³~º bB ²z®ï, *Û, úã®B ~ ~òö º £Ïb . ß® f ´ö jæ" C& ³¦~² F>Ú ®º ©b rJ^ ®b, 7 ÿ§æOö 6Ò ª>Ú ®º ©b rJ^ ® (17,18). F öB æ¾Z~ rÎ ï 5 ÖzÎ"ö & ¢ Û~ æ¾Z òÒ zêR ºÂb~ ÖzÎ"& çúj C²: ® (19). ö Bº * LDLÖzö & æ¾Z ºÂb 5 æ¾Z òÒ ª³b~ ÖzÎ"ö & æÒj áîVö ö ~¶ .. Òò 5 O» bÒò ;öê ;F ³ëVFbV 5 ;Fæ'³»ö B 1999jê 9úö æ¾Z~ , ´, *V, 5 òÒ¢ /Aj ¾² ê.ê, r~ −20oC ïÿö &~B . þö ÒÏ~& . £ Agarose, ascorbate, bromophenol blue, butylated hydroxytoluene(BHT), coomassie brilliant blue R-250, cupric sulfate, 1,1diphenyl-2-picrylhydrazyl(DPPH), 2,4-dinitrophenyl-hydrazine(DNPH), 1,1,3,3-tetra-ethoxypropane(MDA), ethylenediamine tetraacetic acid (EDTA), low density lipoprotein(LDL), sodium dodecyl sulfate (SDS), thiobarbituric acid, trichloroacetic acid, α-tocopherol, f SigmaÒ B®(St. Louis, MO, USA)j «~ ÒÏ~& . Disodium hydrogen phosphate, potassium chloride, potassium phosphate, sodium barbital, sodium chloride, sodium phosphate, f WakoÒ B®j ÒÏ~&, Ca, Cu, Fe, K, Mg, P, Zn ~ &®f ¢ WakoÒ~ B®j ÒÏ~&b, butanol, chloroform, cyclohexane, ethanol, ethyl acetate, heptane, hexane, hydrogen chloride, methanol ~ FVÏ 5 Væ£f 1/£ _f ö &~º ©j «~ ÒÏ~& . ºÂ 5 ª³ æ~ , ´, *V, 5 òÒ¢ ¾² ^. Ê, ªê~ , ~~ï'V¢ ¦O 2¢Êö ò7ï~ 20V~ öê R(ethanol, EtOH), zêR(methanol, MeOH), ª(chloroform, CHCl3), Ò b(water)j &~ >»çöB 24* & NºÂ~& . ®Bbj B~V *~ Whatman No. 2 " æ " r, 6{³»~, ''~ ºÂb ò ÒÏ ~& . Öz W &Ë ¸~~ æ òÒ zêR ºÂ b~ ãÖöº, WWª ßWj ¦Æ~V *~, æ ò Ò zêR ºÂbj bö *çÎ Ê, Ö(n-hexane), ª(chloroform, CHCl3), öj^rÞ(ethyl acetate, EtOAc), ¦êR(n-butanol, n-BuOH) Bb WNö V Ï ª³j >¯ Ê, " r, 6{³»~, áf ''~ ª³bj ò ÒÏ~& . Özbîö & W7ê 5 ¾¦9 zb ï W%ê G;: ¦*ê æ¾Z~ ºÂb 5 òÒ ª³bö &~ ÖzW bî rJê zb (protein, aromatic amine, 5 phenol )~ ÏÂ;ê¢ Hewlett PackardÒ(Palo Alto, CA, USA)~ HP 8452A diode array spectrophotometer¢ ÒÏ~. 285 nmöB 7ê G;~& (20). r òº 0.1 mg/mL& >² ºÂbj zêRö ê, 7ê¢ G;~& . RH ï: Carotenoidsïf ºÂbj 0.01%& > ê zêRö ê, 450 nmöB 7ê¢ G;~ ïj G;~& (21). .ïê: ¦*ê æ¾Z ºÂb 5 òÒ ª³b~ .ïz >w Wbî~ ³ê¢ ¾æÚº .ïêº 490 nmöB~ 7ê¢ ¶F/&7F ª77êê(Hewlett PackardÒ~ HP 8452A diode array spectrophotometer)¢ ÒÏ~ 7ê¢ G ;~& (20). +¾¦9 z> ï: C¾¦W bî ïf Folin-DenisO »ö V¢ ªC~& (22). ¯ ºÂb _f ª³bj 1.0 mg/mL & >² B ê, 75 mL~ Ã~>& FB 100 mL~ zÊ 2¢Êö 1 mLO I ¾ b~ Folin-Denis 5 mLf êÖ¾Þ zÏ 10 mL¢ Nf If r Ã~> 100 mL Ïïb jÚ . ©j ¾ b~ NöB 30ª O~ ê UV/VIS ª77êê 760 nmöB 7ê¢ G;~ &b, &bîf tannic acid¢ Ï~& . LDL~ Özö & Öz 9 LDL~ ªÒ: .öB density 1.025-1.055 g/mLö ~º lipoproteinj áV *~ F human plasmaö 0.1% EDTA f 0.02% NaN3¢ &~ v> ê, KBr(d = 1.006 − 1.025)j &~ 1N öªÒ(4,000 rpm, 5oC, 15 hr) . r ªÒB VLDLj B~ LDL B fractionj ê, KBr (d = 1.026 − 1.055)j &~ 2N öªÒ(40,000 rpm, 5oC, 24 hr)~ LDLj ªÒ~& (16). ªÒ LDLf 0.005 M Tris, 0.05 M NaCl, 0.02% EDTA buffer(pH 7.4) RCÊ ïÿ ~ ÒÏ~& . LDL~ Öz: LDL(65 µg or 130 µg)" CuSO4(20 µM)ö * Ú¦b& 1 mL& >ê phosphate buffer(pH 7.4)¢ Df Ê, 37oCöB 4* >wÎ Ê, EDTA(1 mM)¢ 20 µL Î&~ Öz¢ 7æ8 (12). Thiobarbituric acid reactive substance(TBARS)~ G;: LDLÚ~ Özæîï(MDA~ ·)f TBARSO»b G;~&. (12). ºÂb _f ª³b~ ³êæzö V MDA~ ïæ z¢ G;~&, ª³b~ ¢;³ê(30 µg/mL)öB *(0-8 hr)ö V TBARS~ ïæz¢ G;~ LDLÖzö ~º æ¾Z ºÂb _f òÒ ª³b~ Öz Î"¢ þ~ & . ¯, EDTAÎ& Öz& 7æB LDLÏö TCA-TBAHCl ;æ(15% trichloroacetic acid: 0.375% thiobarbituric acid: 0.25 N HCl) 3 mL¢ &ê, 95oCöB 30ª* 7ûê, ï 'Î Ê, 3,000 rpmöB 10ª* öªÒ ê, çj ~ WB MDA~ ïj 532 nmöB ª77êê¢ Ï~ G;~& . Conjugate diene ï: æOî Özb~ G;j *~ æî Özê¢ conjugated dienes O»j Ï~& (12). LDL Öz Ïö chloroform 2 mLf methanol 1 mL¢ Dfê, 5ª* 1,000 rpmöB öªÒ~& . ç[j ªÒ, j¾[j î.

(4) æ¾Z ºÂb~ ÖzÎ". 313. Table 1. Contents of total phenolics in Rosa davurica Pall extracts Extracts EtOH MeOH CHCl3 Water. Total phenolics (g/100 g) Fruit. Leaf. Stem. Root. 10.1 10.4 00.8 08.8. 9.5 9.4 0.4 8.4. 10.60 9.6 0.6 8.3. 10.60 9.1 1.3 9.5. Table 2. Absorbance of methanol extract and their solvent fractionations obtained from the roots of Rosa davurica Pall at 285 nm, 450nm, and 490 nm. Fig. 1. Absorbance of Rosa davurica Pall extracts by using various solvents at 285, 450, and 490 nm. The absorbance is measured with 0.01% solution in methanol over an optic path of 1 cm.. ²BÊ¢ Ï~ Î ò¢ cyclohexane 3 mLö ª77êê¢ Ï~ 234 nmöB conjugated diene~ ïj G;~& . Apolipoprotein carbonyl ï: LDL ÖzÏ(1 mL)ö 10 mM DNPH 200 µL¢ Dfê, çNöB 60ª* >wÎ Ê, denaturing buffer(150 mM sodium phosphate, 3% SDS, pH 6.8) 0.6 mL¢ Î& Ê, b~& . 1.8 mL reagent alcohol" 1.8 mL heptanej Î&~ Dfê, 3,000 rpmöB 10ª* ö ªÒ~ protein pelletj denaturing buffer 3 mLö Ê, 320-410 nmöB scan Ê, peak absorbance¢ ÒÏ~ protein carbonyls(extinction coefficient = 22,000 M−1cm−1) ïj êÖ~ & (12).. Ö" 5 8 F öB &¶º æ¾Z òÒ zêR ºÂb Öz Î"& Ö çúj C²: ® (19). öBº æ¾Z( , ´, *V, 5 òÒ) ºÂb 5 æ òÒ ª ³bj ÒÏ~ LDL Özö & Öz Î"¢ «~¶ >¯~& .. Fractions. A285. A450. A480. MeOH ext. n-Hexane fr. CHCl3 fr. EtOAC fr. n-BuOH fr. Aqueous fr.. 1.790 0.290 0.377 2.405 1.747 0.222. 0.071 0.028 0.016 0.012 0.056 0.010. 0.051 0.023 0.011 0.008 0.038 0.007. ¦*ê æ¾Z ºÂb~ Özbîö & W7ê 5 ¾ ¦9 zb ï æ¾Z ºÂb~ 285 nm, 450 nm 5 490 nmöB~ Ö zbîö & 7ê¢ Fig. 1ö ¾æÚî . ÖzW bî phenolê zb ~ >& ¢Ú¾º 285 nmöB~ 7êº òÒ~ zêR 5 öêR ºÂb &Ë ¸~b òÒ > ´ > *V > Bb 7ê& ¸~ . Ï êº zêR > ö êR > b > ª Bb 7ê& ¸² G;>î . & ¦ª~ carotenoidsº 450 nmöB >~æ 7ê ïj º; > ® . A450~ 8f 0.009-0.225 Ö Ô² G;>î . æ¾Z ºÂb~ 490 nmöB G; .ïêê 0.006-0.188 Ö Ô² G;>î . *~ Ö" Ú j B 285 nmöB >¢ ¾æÚº bî ï F>Ú ®rj { > ®î . æ¾Z ºÂböBº carotenoids¾ . ïz >w Wb Öz Wö ~º 'Ë 'j © b º;>î . æ¾Z~ C¾¦Wzb ïj Folin-DenisO»ö V¢ ªC Ö"¢ Table 1ö ¾æÚî . ª ºÂbj B ~ Î ¦*~ ºÂböB 100 g 8.3-10.6 gb Ö ¸ f ·j F~ ®î . ß®, æ¾Z òÒ 5 *V~ ö êR ºÂb 10.6 g/100 gb &Ë ¸f ïj æî ®î. . Ö"º *~ 7ê Ö"f ¢~~º Ö"B 285 nm öB >~º ¾¦W zb "B ÒWbî¢ ©b º;>î . æ¾Z òÒ ª³b~ Özbîö & W7ê æ¾Z òÒ ª³b~ Özbî ÏÂ;ê¢ 7ê¢ G;~ ¦Æ~& . Ö"¢ Table 2ö º£~& . Table 2öB º :f ?, rê zb(A450)" .æb î(A490)~ ãÖöº 7ê& ~ ¾æ¾æ pº ©b Ú jB æ¾Z òÒ ª³b~ "B Wbî jò © b 6>Úê . ¾ ¾¦êzb~ ÏÂ;ê¢ ¾æÚ º 285 nmöB~ 7êº öj^rÞ ª³b 2.405. Ö ¸² G;>º ©b jB, ¾¦ê zb Ö ô.

(5) 314. ®"²æ B 36 ² B 2 ^ (2004). Fig. 2. Inhibitory effects of Cu2+-mediated LDL lipid peroxidation by increasing concentration of Rosa davurica Pall extracts as evaluated by TBARS assay. LDL (equivalent to 65 µg protein) was incubated with 20 µM Cu2+ at 37oC under air for 8 h in the presence of varying concentrations of methanol extracts. The reaction mixture was stopped by adding 3 mL of TBA-TCA-HCl solution, and then the solution was heated at 95oC for 30 min. After centrifugation to clarify the solution, the peak absorbance at 532 nm was used to calculate TBARS using malonaldehyde bis (dimethylacetal) as standard. ù--ù: Fruit, þ -þ : Leaf, -- : Stem, ü --ü : Root.. ÏÂ>Ú^ ¾Jº ©j {~& . rb ¦êR ª³böB ¾¦W zb ô ÏÂ>Ú^ ¾Jº ©j { ~&b, ª, Ö 5 b ª³böBº ²ï ÏÂ >º ©j {~& . LDL Özö & ¦*ê æ¾Z ºÂb~ Öz Î" F öB, " æ¾Z zêR ºÂböB DPPH radical ²W ¾æÎj {~& (19). þöBº LDL Özö & æ¾Z( , ´, *V, 5 òÒ) zêR ºÂ b~ Öz Î"¢ MDA8j G;~ ¦ï~& . æ¾ Z zêR ºÂbj 20, 40, 60, 80 µg/mL~ ³ê Î&~& j r~ MDA8j Fig. 2ö ¾æÚî . æ¾Z òÒ zê R ºÂb~ ãÖ, 66.0, 85.3, 97.1, 100%~ MDA W ÛBN j &, æ *V zêR ºÂb~ ãÖöê ³ê& Ã &ö V¢ MDAWj ÛBj ¾æÚî . ¾, æ 5 ´~ zêR ºÂb~ ãÖöº MDA Wj ÛB~ æ á~& . æ òÒ zêR ºÂb~ ãÖ, LDL Özö &~, ²ï~ ³êöBê &Ë ±f Öz Î"¢ ¾æÚæ ê~ þöBº æ òÒ zêR ºÂb¦V Ï. ª³bj B ê, ª³bj ÒÏ~ þ~& . LDL Özö & æ¾Z òÒ ª³b~ Öz Î" LDL Özö & &Ë ±f Öz Î"¢ ¾æÞ æ ¾Z òÒ zêR ºÂbj FVÏ ö V ª³j >¯ Ê, ''~ ª³bj ÒÏ~, MDA, conjugated diene, apolipoprotein carbonyl& j ªC~, LDLö & Öz Î" ¢ G;~& . æ¾Z òÒ ª³b~ ³ê(0-40 µg/mL)ö V MDA8 ~ æz¢ Fig. 3ö ¾æÚî . Ö ª³b" b ª³bj B. Fig. 3. Inhibitory effects of MDA formation on LDL by increasing concentration of each fraction from the methanol extract of roots of Rosa davurica Pall. LDL(equivalent to 65 µg protein) was incubated with 20 µM Cu2+ at 37oC under air for 8 h in the presence of varying concentrations of methanol extracts. The reaction mixture was stopped by adding 3 mL of TBA-TCA-HCl solution, and then the solution was heated at 95oC for 30 min. After centrifugation to clarify the solution, the peak absorbance at 532 nm was used to calculate TBARS using malonaldehyde bid (dimethylacetal) as standard. ø--ø: Hexane fr., ù--ù: Chloroform fr., ü --ü : Ethyl acetate fr., -- : Butanol fr., þ --þ : Aqueous fr.. EtOAc ª³b, BuOH ª³b, Ò CHCl3 ª³b ³ ê& Ã&ö V¢ MDA Wö & ¸f ÛBNj ¾æÚ î . æ¾Z òÒ ª³b 30 µg/mL~ ³êöB BuOHª ³bf 93.2%(2.6 nmol/mg), EtOAc ª³bf 85.3%(5.6 nmol/ mg), Ò CHCl3 ª³bf 84.0%(6.1 nmol/mg)~ ÖzÛBN (MDA8)j ¾æÚî . 30 µg/mL ;ê~ ²ï~ ª³b ³ê öBê LDL Özö & Öz Î"& ;~² ¾æ¾æ ê~ þöBº 8j þ³ê ;~ >¯~& . * ö V æ¾Z òÒ ª³b~ MDA8j Fig. 4ö ¾æÚ î . Hexane ª³b" b ª³bf control" FÒ æzïj ¾æÚº ©b j, *ö V LDL ÖzÛBËK £ ©b ¾æÒ, BuOH ª³b, EtOAc ª³b, Ò CHCl3 ª³bf 8*ræ LDL~ Öz¢ ;K~² ÛBj "î . Ö" Ú " r, BuOH ª³b, EtOAc ª ³b, Ò CHCl3 ª³b~ Ú¦ö Â ÖzBB~ B B &ËW ®º bî Ò~ ®rj ºG > ®î . ª³b 30 µg/mL~ ³ê Î&~&jr~ conjugated diene ï~ ÛB;ê¢ Fig. 5ö ¾æÚî . BuOH ª³b" EtOAc ª³b 5.6V 5 2.2V~ conjugated diene~ W 6²¢ ¾ æÚî, Ö ª³bf ~ ÛB~æ áj "î . ª ³b 30 µg/mL~ ³ê Î& apolipoprotein carbonyl ï Û B;ê¢ Fig. 6ö ¾æÚî . BuOH ª³b 3.4V~ 6² ïj ¾æÚîb, EtOAc ª³b" CHCl3 ª³b 1.5V ; ê~ apolipoprotein crabonyl ï 6²¢ ¾æÚî . Ö ª ³bf *& ÛB~æ á~& . Ú jB BuOH ª ³b, EtOAc ª³b, Ò CHCl3 ª³bö Ò²~ &&ê æ OWî~ Öz¢ ÛB > ®º Â ÖzB& ï Ò~ ®rj {~& ..

(6) æ¾Z ºÂb~ ÖzÎ". Fig. 4. Inhibitory effects of each fraction from the methaol extracts of roots of Rosa davurica Pall according to LDL oxidation time. LDL (equivalent to 65 µg protein) was incubated with 20 µM Cu2+ at 37oC under air for 8 h in the presence of each fraction (30 µg/mL). The reaction mixture was stopped by adding 3 mL of TBA-TCAHCl solution, and then the solution was heated at 95oC for 30 min. After centrifugation to clarify the solution, the peak absorbance at 532 nm was used to calculate TBARS using malonaldehyde bid (dimethylacetal) as standard. þ--þ: Control, ù--ù: Hexane fr., ø --ø: Chloroform fr., ü --ü : Ethyl acetate fr., -- : Butanol fr., ý--ý: Aqueous fr.. Özbîö & W7ê 5 Öz Î"f~ ç&9 ¦*ê æ¾Z ºÂb~ Özbîö & 7ê¢ G ; Ö", ¢¦ ÖzW bî phenolê zb ~ > & ¢Ú¾º 285 nmöB~ 7ê& &Ë ¸~ . ¾ rê zb ~ 7 ¢Ú¾º 450 nmöB~ 7 ê¾ .æbî~ 7 ¢Ú¾º 490 nmöB~ 7êº. Ö Ô² G;>º ©b jB, æ¾Z ºÂböBº " 285 nmöB 7j ¾æÚº bî Öz Wj ¾æ â ©b ÒòB . ¦*êº òÒ~ zêR ºÂb &Ë ¸² G;>îb ´ > *V > Bb 285 nmöB~ 7 ê& ¸² G;>î . $ LDL Özö & ¦*ê æ ¾Z ºÂb~ Öz Î"¢ G; Ö", òÒf ´ zêR ºÂböB ;K MDA Wj ÛB~& . æ¾Z òÒ ª³b ö & Özbîö & 7ê G;Ö"~ ãÖ öê " 285 nmöB >>º ¾¦ê zb ~ ÏÂ & Ë ¸f ©b {>î . ß® EtOAc ª³b 5 BuOH ª ³böB ¸f 7j ¾æÚîb, ª³b LDL Ö zö & ;K Öz Î"¢ ¾æÚî . ç~ Ö" Ú jB, ¦*êº æ¾Z òÒö 285 nmöB 7>º ¾¦ê zb ï F>Ú ®b, bî. " EtOAc ª³b 5 BuOH ª³bö Ò ©b Ò òB . V¢B LDL Özö & æ¾Z ºÂb~ Öz Î"º ¾¦ê zb ö ~ 'Ë¢ ©b ÒòB . Ò²~ &&êæOWî~ Özö & æ¾Z ºÂb ~ Öz Î"¢ G; Ö", æ¾Z òÒ zêR ºÂ b MDAWj &Ë ;K~² ÛBj "î . æ ¾Z ºÂbf " 285 nmöB >¢ ¾æÚº bî ï F>Ú ®b, ¾¦Wzb ïj G; Ö", ª ª³bj B Î ºÂbö ï F>Ú ®î . . 315. Fig. 5. Inhibitory effects of each fraction from the methanol extracts of root of Rosa davurica Pall on conjugate diene formation. LDL (equivalent to 130 µg protein) in PBS pH 7.4 was incubated with 20 µM Cu2+ at 37oC under air for 8 h in the presence and absence of each fraction (30 µg/mL). Conjugate diene formation was measured by the method of Yan et al. (12). Fig. 6. Inhibitory effects of each fracion from the methanol extracts of root of Rosa davurica Pall on Cu2+-mediated apoB carbonylation. LDL (equivalent to 65 µg protein) in PBS pH 7.4 was incubated with 20 µM Cu2+ at 37oC under air for 8h in the presence and absence of each fraction (30 µg/mL). Conjugate diene formation was measured by the method of Yan et al. (12). æ¾Z òÒ zêR ºÂb Ò²~ &&êæOWî Ö zö & Öz Î"& &Ë ;K~² ¾æÒV r^ö, ò Ò zêR ºÂbj z ;B~V *~ Wö V Ï. ª³bj Bê, ''~ ª³bj &çb LDL Özö & Öz Î"¢ G;~& . æ¾Z öj^rÞ ª³b 5 ¦êR ª³b MDA, conjugate diene, apolipoprotein carbonyl& ~ ªCÖ" LDL Özö & Öz Î" & Ö çúj {~& . $ æ¾Z òÒ~ öj ^rÞ ª³b 5 ¦êR ª³bf 285 nmöB >>º ¾ ¦W zbj ï F~ ®rj {~& . $ F öB ª³böº ï~ catechin F>Ú ®rj : ® (19). ç~ Ö"¢ «~ æ¾Z òÒ.

(7) ®"²æ B 36 ² B 2 ^ (2004). 316. ~ öj^rÞ 5 ¦êR ª³b~ "B Wbîf catechin " ?f 2¢ê zb" ï~ ¾¦W zb¢ ©b º;B .. º. £. jæ"-C¢ ï F~ ®º æ¾Z~ ¶öz¢ * B ¾¦W zb~ ï 5 Ò²~ &&êæOW î~ Özö & Öz Wj G;~& . æ¾Z ºÂ b~ ¾¦W zb ïj G; Ö", Î ¦*öB 100 g 8.3-10.6 gb Ö ¸f ·j F~ ®î . $, æ¾Z ºÂb 5 ª³bö & Öz bî~ ïj G; Ö", æ¾Z òÒ zêR ºÂb" òÒ~ öj^r Þ 5 ¦êR ª³b 285 nmöB >¢ ¾æÚº ¾¦W zbj ï F~ ®rj {~& . TBARSö ~ LDLÖz ÛBÎ" G;, æ¾Z òÒ~ zêR ºÂb &Ë ±f Î"¢ ¾æÚî . æ¾Z òÒ~ öj^ rÞ 5 ¦êR ª³b~ ãÖ, 30 µg/mL Î& 85.3% 5 93.2%ræ LDL Öz¢ ÛBj {~& . $ CHCl3 ª ³b, EtOAc ª³b, Ò BuOH ª³bf 30 µg/mL ª³b Î& Cu2+ö ~ LDL Öz¢ 8*ræ ÛBj {~ & . Cu2+ö ~ apoB carbonylation" conjugate diene;W Û BÎ" G;, EtOAc ª³b" BuOH ª³b &Ë çú~ ² ÛBj "î . æ¾Zº ï~ ¾¦W zb j F~ ®b Öz W Ö çú~æ îÚ V ËW® ²Ò~ Ï V&>º £Ïb¶ö .. 6Ò~ º ;Fæ'³» 5 ;F³ëVFbV ¦V ÏÒë æö(2000. 1.-2001. 12.)ö ~ Úê Ö"B ö 6Òãî .. ^. ò. 1. Caslake MJ, Packard CJ, Shepherd J. Plasma triglyceride and LDL metabolism. Eur. J. Clin. Invest. 22: 96-104 (1992) 2. Henriksen T, Mahoney EM, Steinberg D. Enhanced macrophage degradation of biologically modified low density lipoprotein. Arteriosclerosis 3: 149-159 (1983) 3. Morel DW, Docorleto PE, Chisolm GM. Endothelial and smooth muscle alter low density lipoprotein in vitro by free radical oxidation. Atherosclerosis 4: 357-364 (1984) 4. Bruckdorfer KR. Free radicals, lipid peroxidation and atherosclerosis. Curr. Opin. Lipidol. 1: 529-535 (1990) 5. Steinbrecher U, Parthasarathy S, Leake DS, Witztum JL, Steinberg D. Modification of low density lipoprotein by endothelial cells involves lipid peroxidation and degradation of low density. lipoprotein phospholipids. Proc. Natl. Acad. Sci. USA 83: 38833887 (1984) 6. Esterbauer H, Streigel G, Puhl H, Oberreither S, Rotherneder M, El-Saadani M, Urgens G. The role of vitamin E and carotenoids in preventing oxidation of low density lipoproteins. Ann. N. Y. Acad. Sci. 570: 254-267 (1989) 7. Manigiapane H, Thomson J, Salter A, Brown S, Bell GD, White DA. The inhibition of the oxidation of low density lipoprotein by (+)-catechin, a naturally occurring flavonoid. Biochem. Pharmacol. 43: 445-450 (1992) 8. De Whalley CV, Rankin SM, Hoult JRS, Jessup W, Leake DS. Flavonoids inhibit the oxidative modification of low density lipoproteins by macrophages. Biochem. Pharmacol. 39: 1743-1750 (1990) 9. Machlin LJ, Bendich A. Free radical tissue damage: Protective role of antioxidant nutrients. FASEB J. 1: 441-445 (1987) 10. Retsky KL, Frei B. Vitamin C prevents metal ion-dependent initiation and propagation of lipid peroxidation in human low-dinsity lipoprotein. Biochim. Biophys. Acta 1257: 279-287 (1995) 11. Noguchi N, Gotoh N, Niki E. Dynamics of the oxidation of low density lipoprotein induced by free radicals. Biochim. Biophys. Acta 1168: 348-357 (1993) 12. Yan LJ, Dray-Lefaix MT, Packer L. Ginko biloba extract(EGb 761) protects human low density lipoproteins against oxidative modification mediated by copper. Biochem. Biophy. Res. Com. 213: 360-366 (1995) 13. Jeong J, Kim E, Hwangbo H, Ham S. Effects of Ligularia fischeri extracts on oxidation of low density lipoprotein. Korean J. Food Sci. Techol. 30: 1214-1221. (1998) 14. Park CO, Kim KS, Ji YA, Ryu BH. Antioxidant activity of daidzin and puerarin toward oxidation of human low density lipoprotein. J. Korean Soc. Food Sci. Nutr. 26: 25-31 (1997) 15. Park CO, Jin SH, Ryu BH. Antioxidant activity of green tea extracts toward human low density lipoprotein. Korean J. Food Technol. 28: 850-858 (1996) 16. Yag KS, Jeon CM. Effect of Taraxacum coreanum Nakai on low density lipoprotein oxidation. Korean J. Pharmacogn. 27: 267-273 (1996) 17. Shin KH, Chung HS, Cho SH. Vitamin contents in the fruits of Rosa davurica Pall. J. Med. Crop Sci. 3: 21-24 (1995) 18. Shin KH, Lim SS, Lee SH, Seo JS, Yu SY, Park CH. Vitamin content in Rosa davurica Pall. J. Medicinal Crop Sci. 6: 6-10 (1998) 19. Sa JH, Shin IC, Jeong KJ, Shim TH, Oh H, Park SK, Cheung E, Kim SN, Kim GG, Choi DS, Kwon YS, Kim CM. Catechin content and antioxidative effect from Rosa davurica Pall. Korean J. Pharmacogn. 33: 177-181 (2002) 20. Kim JY, Maeng YS, Lee KY. Antioxidative effects of soybean extracts by using various solvents. Korean J. Food Sci. Technol. 27: 635-639 (1995) 21. Seo YH, Kim IJ, Yie AS, Min HK. Electron donating ability and contents of phenolic compounds, tocopherols and carotenoids in waxy corn (Zea mays L.). Korean J. Food Sci. Technol. 31: 581585 (1999) 22. AOAC. Official Methods of Analysis, 15th ed. Association of Official Analytical Chemists, Arlington, VA, USA (1980) (2003j 9ú 3¢ 7>; 2004j 4ú 13¢ j).

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