The Effect of Rubus coreanum Miquel Against Lipopolysaccharide-induced Oxidative Stress and Lipid Metabolism

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1 J. Fd Hyg. Safety Vol. 22, No. 3, pp. 213~217 (2007),QWTPCN QH (QQF *[IKGPG CPF 5CHGV[ Available online at äæ Ú LPSÊ ò È ò Ü ²Â öâ Î Â 1  ý 1 Â ß * w w w œw, 1 w The Effect of Rubus coreanum Miquel Against Lipopolysaccharide-induced Oxidative Stress and Lipid Metabolism In Deok Kim, Kum Suk Kang, Ryun Hee Kwon, Jeong Ok Yang 1, Joong Sook Lee1, and Bae Jin Ha* Department of Pharmaceutical Engineering, College of Medical Life Science, Silla University, Busan , Korea 1 Department of Physical Therapy, College of Medical Life Science, Silla University, Busan , Korea (Received July 16, 2007/Accepted August 26, 2007) ABSTRACT LPS induces the synthesis of several inflammatory cytokine, chemokine, NO and inflammation in the liver of rats. The purpose of this study was to investigate the preventive effects of Rubus coreanum Miquel (RCM) in lipid metabolism. RCM of 100 mg/kg concentration was intraperitoneally administered into rats at dose of 1.5 ml/kg for 20 days. On the day 21, 1.5 ml/kg of LPS was injected 4 hours before anesthetization. We examined the lipidrelated functions by measuring the levels of triglyceride (TG), total cholesterol (TC), total lipid (TL), high-density lipoprotein cholesterol (HDL-C) in serum and malondialdehyde (MDA) in liver tissue. The results showed that LPS treatment increased the values of TG, TC, TL and MDA, decreasing that of HDL-C. But RCM pretreatment decreased the high values of TG, TC, TL and MDA to the low values and increased the low value of HDL-C to the high value. These results suggested that RCM could be used as the potential candidate for the lipid metabolism natural supplement. Key words: Rubus coreanum Miquel, lipopolysaccharide, MDA, HDL-C, Total lipid õ ù w e, 2 w y ùkü w ƒ y w š. (Rubus coreanum Miquel) ù û w, š. l w»,,,, x w yw z ƒ š š 1,2). ù w» l tannin flavonoids yw š 3-5), gallic acid, 2,3-(s)-HHDP-D-glucopyyranose, sanguin š 6). *Correspondence to: Bae Jin Ha, Department of Pharmaceutical Engineering, College of Medical Life Science, Silla University, 1-1 San, Gwaebup-dong, Sasang-gu, Busan, , Korea Tel: , Fax: bjha@silla.ac.kr Ê w w w w B s T s w š 7). y p j Lipopolysaccharide (LPS) ³ s w. s(mø)ù w y w cytokine, w, v y w ù w» w 8). LPS-binding protein w MØ CD14 qx jƒ ù» inducible nitric w 9), oxide synthase y y g NO ƒ k 10). NO MØ y w oxygen radical w w w NOƒ 2+ Ca ƒ w superoxide anion (O 2 ) H 2 O 2ƒ wì y w - OH g s w y w 11). w radicals s œ w»wš12), mw s q ƒ 13). y p k š w ƒ. 213

2 214 In Deok Kim et al. w y ƒ š. y ƒ DNA w ¾ œ y p y (lipid peroxide), y (oxidized protein) (mutation) w y w. 14,15) ƒ w ƒ x, LDL-g l w wg l (anti-cholesterol factor) HDL-cholesterol x š. 16,17) ƒ z y w w ƒƒe š w LPS l y p w z x w y t w z š w. Ì Ý ßá Ì s y w w. µú Ý x 170~180 g ü z 7 f (Sprague-Dawley) z l œ š 7 g. x ù ù š cage jš šx w w (Table 1). Auto Control System»»(SS-2000, daejong, Korea) w 22±1 C o 60±5% g. (NOR; normal group) ml/kg of 0.9% saline n w. (CON; LPS-treated group) mg/kg of 0.9% saline n w z 21 ú LPS 5 mg/kg 1 ml/kg ü n w. w (RCM; Rubus coreanum Miquel extract and LPS-treated group) 100 mg/kg 1.5 ml/kg n w z 21 ú LPS Table 1. Experimental design of rats 5 mg/kg 1 ml/kg ü n w. x y p g. LPS n wš k 4 z ether wš g x wš w xw. äæ Ì 100 g 1 L y þ ƒ mw 8 w evaporator w w w x w. Ý n» z x ether w w z l w ƒ x xq ¼ z, 30 g»(mega17r, Hanil, Korea) w x w x w w. 4 w 0.9% w z -70 o C deep freezer w x w. 5 PBS (ph 7.4) ³ yw z 800 G, 4 C o 10 min w x w. ßá Triglyceride (TG) É x p (triglyceride) w TGkp (Triglyceride kit, Young Dong, Korea) w w. x» mw x 10 ul w kit mw w d w. Total Lipid É x w d w x 0.1 ml H 2 SO 4 2 ml ƒw z 100 C o 10 ƒ w. ƒ z þƒ eš 100 ul w phospho-vanillin reagent 5 ml ƒw. 37 C 15 ƒ e z o ŸŸ (Uvikonxs, Bio-Tek, Italy) w 540 nm Ÿ d w. Total-cholesterol ë É x g l w d kp (cholesterol kit no. BC Experimental day 1-20 day 21 group dose of sample dose of sample NOR (7) 1.5 ml/kg of 0.9% saline, i.p 1.5 ml/kg of saline, i.p. CON (7) 1.5 ml/kg of 0.9% saline, i.p 1.5 ml/kg of LPS 1.5 ml/kg of Rubus coreanum RCM (7) (5 mg/kg), i.p. Miquel fraction (100 mg/kg), i.p. NOR : normal group, CON : LPS-treated group, RCM : Rubus coreanum Miquel-pretreated and LPS-posttreated group. The number of experiment animals is given in parenthesis. i.p. : intraperitoneally, LPS : Lipopolysaccharide.

3 The Effect of Rubus coreanum Miquel Against Lipid Metabolism E, Young Dong, Korea) w w. x» mw x 10 ul w kit mw w d w. HDL-cholesterol É x HDL-g l w d kp (HDL-cholesterol kit no. A308-HDL, Young Dong, Korea) w d w. x e g 10 e z 3000 rpm e k z w x» mw w d w. Malon dialdehyde(mda) É y p w t Ha 18) d w. ŸŸ (Uvikon-xs, Bio-Tek, Italy) 535 nm q Ÿ d w. Ð x w x s³e t r ùkü š, m SPSS w ANOVA w. ŠŒ Ý Triglyceride (TG) Ü TG ü xk w (chronic degenerative disease) wù. w TG ü w ƒ w ƒ». x n wš LPS zn w TG w yz. TG w e z Fig. 1 ùkü. LPS n w ƒ ù LPS n w w TG w 3.9 ƒw š, n w 56.4% z ùkü l w z ƒ. Total Lipid Ü Total lipid x w d w w š x, y, k. Fig. 2 w 1.67 w ƒw. w ƒ ù, RCM n w 59.2% z w. Total-cholesterol Ü Total cholesterol ù. x cholestrol ƒ, y, šx x y w k. n w x g l w w Fig. 3. g Fig. 2. Effects of RCM fractions on total lipid activities in LPStreated rat. NOR: normal group, CON: LPS-treated group, RCM: Rubus coreanum Miquel-pretreated and LPS-posttreated group. ***p<0.001, *p<0.05 values are mean ± SE (n=7) Fig. 1. Effects of RCM fractions on TG activities in LPS-treated rat. NOR: normal group, CON: LPS-treated group, RCM: Rubus coreanum Miquel-pretreated and LPS-posttreated group, ***p< value are mean ± SE (n=7). Fig. 3. Effects of RCM fractions on TC activities in LPS-treated rat. NOR: normal group, CON: LPS-treated group, RCM : Rubus coreanum Miquel-pretreated and LPS-posttreated group. ***p< value are mean ± SE (n=7).

4 216 In Deok Kim et al. l d w 1.74 g l w ƒ w w. n w 57.4% w l z ùkþ. HDL-cholesterol (HDL-C) Ü g l g l HDL (high density lipoprotein) - g l x e w y wš x ù w g l. Fig. 4 LPS n w 2.62 w. HDL g l x w g ƒ k. w RCM n w w 81.6% z z. Malondialdehyde (MDA) Ü y ƒ yw w k w x ù ùkü Fig. 4. Effects of RCM fractions on HDL-C activities in LPStreated rats. NOR: normal group, CON: LPS-treated group, RCM : Rubus coreanum Miquel-pretreated and LPS-posttreated group. ***p<0.001 value are mean ± SE (n=7). Fig. 5. Effects of RCM fractions on MDA activities in LPStreated rats. NOR: normal group, CON: LPS-treated group, RCM : Rubus coreanum Miquel-pretreated and LPS-posttreated group. ***p<0.001, *p<0.05 values are mean ± SE (n=7). ƒ w» š,,,, y ƒ w w. w y MDA ƒ x d w. x y x w w š š š w y w w ƒ 19-22). Fig. 5 z ƒ ùkûš, x g l w ùkû. MDA y w 1.74 ùkû, LPS w y y w. n RCM w 57.4% z ü y w z. (Rubus coreanum Miquel) ù û w, š. l w, k,,,, š wš. l w»,,,, x w yw z ƒ š š. LPS ³ s NO jš 2+ Ca ƒ j free radicals x w q wš jš y w w w. y p l z» w,, (RCM) w 20 ü n wš 21 ú LPS 4 n w ƒ w w w y x z y sƒw. (TG) xk w. w. w w ƒ j w. w 3.9 ƒw,. w RCM n w 56.4% w z. (Total lipid) x w d w w š x, y, k. w 1.67 ƒ w RCMn w 59.2% w z.

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