Antioxidant Activity of Momordica charantia L. Extracts

Transcription

1 ª Ÿ (Korean J. Medicinal Crop Sci.) 15(1) : (2007) wy y *Á **Á***Áw****Áxy** *w ww œw, **w ww w, ***w w ww, **** w œw Antioxidant Activity of Momordica charantia L. Extracts Yeol Park*, Hee Ock Boo**, Young Lan Park***, Dong Ha Cho****, and Hyun Hwa Lee** *Department of Biotechnology, Collage of Nature Science,Chosun University, 375 Seosuk-Dong, Korea. **Department of Biology, Collage of Nature Science, Chosun University, 375 Seosuk-Dong, Korea. ***Department of Anatomy, Collage of Medicine, Chosun University, 375 Seosuk-Dong, Korea. ****School of Bioscience & Biotechnology, Kangwon National University, Chunchon , Korea. ABSTRACT : This study was carried out to determine the antioxidant activity and vitamin C contents in plant extracts of the Momordica charantia L. The vitamin C was detected as the highest content in immature fruit (92.2 /100 g), while the content in stem (2.5 /100 g) was lower 40 times than that of immature fruit.gantioxidant activity for the dried sample was investigated by TBA method. The lowest TBARS values were obtained from extracts of dried leaf and followed by ascorbic acid and BHT, showing that the extracts from dried leaf possess the strongest antioxidant activity. Compared with fresh tissues, SOD activity, ATX activity and CAT activity were high level in the dried tissue. These results suggest that the Momordica charantia L. would be a promising antioxidant source as an alternative antioxidant, based on natural plant resources. Key WordsG:GMomordica charantia L., vitamin C, TBA-reactive substance, superoxide dismutase, ascorbic peroxidase, catalase ù q ù, w p w ü y k. w y y, s w, q, y ü w y w w w (Allen et al., 1997; Asada, 1999). ù w y w» w w xk wy wwš p, wy z x w w w (Heath, 1987; Kang et al., 2003). wy z superoxide dismutase (SOD) superoxide anion radical (Á0 2 ) w hydrogen peroxide (H 2 O 2 ) yj z metal cofector w 3, CuZnSOD s, MnSOD mg, FeSOD ƒƒ s. w SOD w y p w ƒ w w, s ü w y x (Bowler et al., 1992). Catalase (CAT) peroxisome w H 2 O 2 ww. CAT H 2 O 2 w w z w ascorbate peroxidase (APX), mg, s s w, ascorbate yk H 2 O 2 yyk (Kang et al., 2003). wr, ü SOD, APX š CAT wy z, tocopherol, ascorbic acid (vitamin C), catechin, glutathion wy, BHT, BHA w wy ù w mw y l yw w. w wy y ƒ y wš wy w y w. x (Momordica charantia L.) 1, úl w w w w Corresponding author: (Phone) ( ) papaya@chosun.ac.kr Received January 20, 2007 / Accepted January 31,

2 wy y (Beloin et al., 2005). ù w ù» wù. ù, ù mw ƒ w, û, vv, w» š (Giron et al., 1991; Park, 1989; Virdi et al, 2003; Zong et al., 1995). p k C, k A w œ š (Grover et al., 2004). l, p, p, rp x w» kw, p w charantin βs w w x û w w šš (Schmourlo et al., 2005; Pakash et al., 2002; Rathi et al., 2002). MAP3 s q w natural killer s yy k s w zƒ d š, momorcharoside DNA, RNA w ww š w z»w š (Bourinbaiar and Lee-Huang, 1995a; Hamato et al., 1995). w in vitro herpes simplex virus-1 poliovirus 1 Candida albocans Crytococcus neoformans y j q w w³y wy z šš (Hamato et al., 1995; Schmourlo et al., 2005). w Á z š, ü w» š, y z w w. wy y w wy ƒ wì jš w. 1. x w (Momordica charantia L.) qw 7-8 yw. ƒ w z 75æ þš w. ƒ w 1 j mk z ƒ 200 g 95% methanol 2 L 24 w. 50æ w z g. 2. k C w d 50 5% metaphosphoric acid wš Whatman No. 1 k, 2,6-dichlorophenol-indolphenol 1, 2% thiourea 2, 2,4 dinitrodphenylhyrazine 1 yww z 50æ 1 g.» y 2,4 dinitrodphenylhyrazined š 30 k z 540 Ÿ dw. 2. x z 4æ þš n w. TBA x w BHT 1% ƒwš 75æ 1 kƒ z þƒg. z 5æ 14 w x w. 3. TBARS d Witte (1970) 10 g 20% trichloroacetic acid 25 ƒw 2 14,000 rpm ³y jš ƒw 100 w w (Whatman No.1) w. w 5 w 2-TBA (0.005 M in water) 5 yww z 15 ewš UV-VIS Spectro photometer 530 q Ÿ dw. TBARS œ w w. TBA (MDA /1000 g) = Ÿ wy z y d 1) z extraction buffer (50 mm phosphate buffer, ph 7.0; 1% Triton X- 100; 1% PVP-40) 1:4 yww z ³yjš g 20 w wy y d w. BSA t w Bradford (1976) dw. 2) Superoxide dismutase (SOD) yd SOD y Beauchamp and Fridovich (1971) 50 mm carbonic buffer (ph 10.2), 0.1 mm EDTA, 0.1 mm Xanthine, mm nitroblue tetrazolium š sw 25æ 10 k z xanthine oxidase (3.3Á10 6 mm) ƒwš 550 NBT Ÿy dw. 57

3 Á ÁÁwÁxy 3) Ascorbate peroxidase (APX) yd APX y Nacano and Asada (1981) 50 mm potassium phosphate (ph 7.0), 0.5 mm ascorbate, 0.1 mm H 2 O 2, 0.1 mm EDTA ƒw 37æ 5 k z Ÿ y dw. 4) Catalase (CAT) yd Catalase y Aebi (1984) 50 mm potassium phosphate (ph 7.0) 10 mm H 2 O 2 ƒw z Ÿ y w. 1 1uM H 2 O 2 ww z 1 unit w. š 1. k C w w SOD, CAT, APX k C, k E š glutathione sww wy yw k e z ew, mw w» ú ƒš (Ji, 1996). k C k w y p w wy w ww,»l yw y w z w w z yy w (Feri et al., 1991). w, y s y y yw, k» z w w (Jakeman, 1993). Table 1 k C w dw 92.2 /100 g 8.41 /100 g w ùkü.» 2.5 /100 g w. ù 77.3 /100 g w ùkü. k C t k C w t tš, š, v,, e j,,», mm. w k C w r, 100 g 5, mm 13-44,» 80 (Park, 1989), k C w. û wš,» k C w w ƒ. wr, (1989) m k C w w m /100 g w yw Table 1. Content of vitamin C in Momordica charantia L. (cv Dragon ) Samples Vitamin C (/100 g) * Immature fruit 92.2 ± 0.03 Mature fruit 84.1 ± 0.09 Leaf 77.3 ± 0.01 Stem 2.50 ± 0.20 * Data represents the mean values ± SE of three independent experiments Fig. 1. Time course changes of TBARS values of in Momordica charandia L. extracts., ù w ƒ t yw wƒ vw. 2. TBARS d, p sy (O 2 ) w y ùš w y ù. w y w aldehyde» malnodialdehyde (MDA) thiobarbituric acid (TBA) w. ƒ» wy wš TBA reactive substane (TBARS) dw (Fig. 1). 0 TBARSe ƒ 0,0997ù, w , eƒ š, wy ascorbic acid» ú û e ùkü wy z ùkü. ù, w w BHT w w asorbic acid w û y ùkü. 58

4 wy y Fig. 2. SOD activity in dry sample (A) and fresh sample (B) in Momordica charandia L. ƒ TBARS e w» w eƒ wy y û. ù 0 TBARSeƒ 0,097ù, w , û e ùkü. wy ascorbic acid e w wy. 3. wy z y šy w» p w y ƒ, wy w (Alsher and Hess, 1993). s s ywš yy k w» w zw zw l š. zw l wy ascorbic acid, glutathione, a-tocoperol, carotenoids š, z l superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase z. SOD, APX š CAT wy z y dwš ƒ w œ w y dw. w SOD y w (139 U/ protein)» (63 U/ protein) ƒ y ùkü. p» w 2 y Fig. 3. APX in dry sample (A) and fresh sample (B) in Momordica charandia L., TBARS e ƒ ùkù wy z. ù, l SOD dw»ƒ 76.5 U/ protein 28.5 U/ protein w 3 ƒ y ùkþ, y (Fig. 2). wy z š SOD y y ùkù ù, y ùkü (Park et al., 1995)., w ƒ, yw yw SOD y ùkü ƒ. APX s w ƒ w w w. y» ascorbic acid w, GPX (glutathione peroxidases)ƒ H 2 O 2 wš y w w wš x (Wheeler et al., 1998). APX y U/ protein ƒ, 8.75 U/ protein», w y ùkü. wr APX y w 12 y ùkü (Fig. 3). CAT ü ww w s yw wy z APX wì H 2 O 2 w w t z. 59

5 Á ÁÁwÁxy 92.2 /100 g ƒ ùkû,» 2.5 /100 g 40 w. ƒ k TBA wyz dw 0,05 ƒ wy z ùkþ, ascorbic acid BHT wy w. w ƒ SOD, APX š CAT y w ƒ y ùkû. w ƒ wy ƒe š w ƒ w š w w w. LITERATURE CITED Fig. 4. CAT activity in dry sample (A) and fresh sample (B) in Momordica charandia L. CAT y ƒ ùkû, 5.7 U/ protein 1.2 U/ protein ùkû (Fig. 4). APX w, ƒ w 5 y ùkü. ƒ wy z z y w, þ y ùkþ. xw w» w»w, û ùsz m z (Bourinbaiar and Lee-Huang, 1995a; Virdi et al., 2003). Yeh (2005)» ¼ CAT SOD» wš yw. x wy ùkþ, p y ù kü., wy wy ƒ wš w. wy y k C w w. ƒ k C w Aebi HE (1983) Catalase: in methodes of enzymatic analyses (Bergmeyer, H.U. ed.). Verlag Chemie. Weinheim. 3: Allen RD, Webb RP, Schake SL (1997) Use of transgenic plants to study antioxidants defense. Free Rad. Biol. Med. 23: Alscher RG, Hess JL (1993) Antioxidants in higher plants. CRC Press, Boca Raton, Asada K (1999) The water-water cycle in chloroplasts: Scavenging of active oxygen and dissipation of excess photons. Annu. Rev. Plant Physiol. Plant Mol. Biol. 50: Beauchamp C, Fridovich J (1971) Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44: Beloin N, Gbeassor M, Akpagana K, Hudson, J, De Soussa K, Koumaglo K, Arnason JT (2005) Ethnomedicinal uses of Momordica charantia (Cucurbitaceae) in Togo and relation to its phytochemistry and biological activity. Journal of Ethnopharmacology 96(1-2): Bourinbaiar AS, Lee-Huang S (1995a) Acrosin inhibitor, 4'- acetamidophenyl 4-guanidinobenzoate, an experimental vaginal contraceptive with anti-hiv activity. Contraception 51(5): Bourinbaiar AS, Lee-Huang S (1995b) Potentiation of anti-hiv activity of anti-inflammatory drugs, dexamethasone and indomethacin, by MAP30, the antiviral agent from bitter melon. Biochemical & Biophysical Research Communications 208(2): Bowler C, Van Montagu M, Inze D (1992) Superoxide dismutases and stress tolerance. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43: Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: Chon SU, Kim, YM, Han SK, Choi SK (2004) Antioxidative 60

6 wy y effects of several compositae plants. Kor. J. Plant Res. 17(1): Feri D, Ames BN (1991) Ascorbic acids as protects plasma lipids against oxidative damage. Nutrition and Cancer 15: Giron LM, Freire V, Alonzo A, Caceres A (1991) Ethnobotanical survey of the medicinal flora used by the Caribs of Guatemala. Journal of Ethnopharmacology 34(2-3): Grover JK, Yadav SP (2004) Pharmacological actions and potential uses of Momordica charantia: a review. Journal of Ethnopharmacology 93(1): Hamato N, Koshiba T, Pham TN, Tatsumi Y, Nakamura D, Takano R, Hayashi K, Hong YM, Hara S (1995) Trypsin and elastase inhibitors from bitter gourd (Momordica charantia LINN.) seeds: purification, amino acid sequences, and inhibitory activities of four new inhibitors. Journal of Biochemistry 117 (2): Heath RL (1987) The biochemistry of ozone attack on the plasma membrane of plant cells. Adv. Phytochem. 21: Jakeman MJ, Edwards RH, Symonns MC (1993) Electron spin resonance studies of intact mammalian skeletal muscle. Biochem. Biophy. Acta. 874(2): Ji LL, Fu R, Mitchell EW (1992) Glutathione and antioxidant enzymes in skeletal muscle: effect of fiber type and exercise intensity. J. Applied Physiology 73(5): Kang NJ, Kwon JG, Lee HC, Jeong HB, Kim, HT (2003) Antioxidant Enzymes as Defense Mechanism against Oxidative Stress Induced by Chilling in Cucurbita ficifolia Leaves. J. Kor. Sod. Sci. 44(5): Nakano Y, Asada K (1981) Hydrogen peroxide os scavenged by ascorbate-specific peroxidase in spinach chloroplast. Pland Cell Physiol. 22: Parkash A, Ng TB, Tso WW (2002) Purification and characterization of charantin, a napin-like ribosome-inactivating peptide from bitter gourd (Momordica charantia) seeds. Journal of Peptide Research 59(5): Park PS (1989) Investigation of Vitamin C in the Lines of Bitter Gourd(Momordica Charantia L.) J. Chinju Nat. Agri. & For. Jr. Coll. 24: Park YJ, Kang M., Kim JI, Park OJ, Lee MS, Jang DJ (1995) Changes of Vitamin C and Superoxide Dismutase(SOD)-like Activity of Persimmon Leaf Tea by Processing Method and Extraction Condition. Kor. J. Food Sci. Technol. 27(3): Rathi SS, Grover JK, Vats V (2002) The effect of Momordica charantia and Mucuna pruriens in experimental diabetes and their effect on key metabolic enzymes involved in carbohydrate metabolism. Phytotherapy Research 16(3): Schmourlo G, Mendonca-Filho RR, Alviano CS, Costa SS (2005) Screening of antifungal agents using ethanol precipitation and bioautography of medicinal and food plants. Journal of Ethnopharmacology 96(3): Virdi J, Sivakami S, Shahani S, Suthar AC, Banavalikar MM, Biyani MK (2003) Antihyperglycemic effects of three extracts from Momordica charantia. Journal of Ethnopharmacology 88(1): Wheeler GL, Jones MA, Smirnoff N (1998) The biosynthetic pathway of vitamin C in higher plants. Nature 393(6683): Witte VC, Krause ME, Bailey T (1970) A new extraction methord for determining 2-thiobarbituric acid values of pork and beef during storage. J. Food Sci. 35: Zong RJ, Morris L, Cantwell M (1995) Postharvest physiology and quality of bitter melon (Momordica charantia L.). Postharvest Biology and Technology 6(1-2):