Materi 8 Produk Indigenous Sebagai Pangan Fungsional Fransiska Rungkat Zakaria Body needs both omega 3 and 6 Linoleic linolenic Jenis Asam Lemak Minyak Kelapa Minyak Kelapa Minyak Inti Sawit (%)a Sawit (%)b (%)b Asam Kaprat 4,9 – 9,5 Asam Kaprilat 5,5 – 9,5 - 3–4 Asam Kaproat 0,0 – 0,8 - 3–7 Asam Laurat 44,0 – 52,0 46 – 52 Asam Miristat 13,0 – 19,0 1,1 – 2,5 14 – 17 Asam Palmitat 7,5 – 10,5 40 – 46 6,5 – 9 Asam Palmitoleat 0,0 – 13,1 Asam Stearat 1,0 – 3,0 3,6 – 4,7 1 – 2,5 Asam Oleat 5,0 – 8,0 39 – 45 13 – 19 Asam Linoleat 1,5 – 2,5 7 – 11 0,5 - 2 Asam Arakhidonat 0,0 –0,4 DEFINISI Pangan tradisional BERBAHAN LOKAL: Bahan baku atau resep makanan dan minuman yang terbuat dari bahan-bahan yang terdapat di Indonesia dan telah dikenal sejak dahulu. • Contoh: tempe, tahu, oncom, bir pletok, wedang jahe, cincau, tape beras, peujeum, dodol, kredok, urap, asinan, sayur asin, kacang rebus, tauge sayuran dan buahbuahan tropis, bahan lalapan, rumput laut, dsbnya Pangan fungsional Makanan atau minuman yang berpengaruh positif terhadap kesehatan, Kegiatan fisik dan mental, disamping kandungan zat-zat gizinya Pangan fungsional/functional food Pharmafoods Pangan tradisional Pangan rekayasa/deigner food Nutraceuticals Fungsional/ Biologis ? Fungsional: Fungsi biologis seluler: sintesis protein, enzim, hormon, DNA, RNA, senyawa metabolik Jenis sel: endotelium, hati dan organ-organ lain, darah merah, imun, syaraf, kulit, penglihatan, dsbnya Sistem: vaskuler, imun, hormon, syaraf, pencernaan regenerasi/pertumbuhan sel (kanker), energi Kesehatan: Tanggung jawab bersama Ilmu dan teknologi pangan dengan farmakologi dan kedokteran Paradigma sehat SAKIT MENJADI SEMBUH = SEHAT OBAT = TIDAK SAKIT TETAP TIDAK MAKANAN (FUNGSIONAL) Pendahuluan: Hubungan antara makanan dan tubuh Eat What Your Body is Made For Pangan/bahan alami: segar, .olahan Metabolisme (pembongkaran) Cells dlm organ tubuh WHO 2002 PARADIGMA SEHAT SEHAT USAHA PENCEGAHAN: PANGAN, NATURAL PRODUCTS Perawatan Pencegahan SEHAT USAHA KEDOKTERAN FARMASI Pengobatan SAKIT It is abundantly clear that the incidence of all the common cancers in human is being determined by various potentially controllable external factors. This is surely the most comforting fact to come out of all cancer research, for it means that cancer is, in large part, a preventable diseases. World Cancer Research Funds (WCRF) & American Institute for Cancer Research (AICR) 1997: 670 halaman Pangan/makanan/bahan nabati/natural products yang dapat mencegah kanker dapat mencegah penyakit jantung, pembuluh darah, diabetes, dan penyakit degeneratif lainnya Pangan/makanan yang dapat mencegah kanker Berbagai jenis sayuran Klorofil serat Daun cincau Antosianin Flavonoid/fenolik Isotiosianat Karotenoid, terpenpoid Bagaimana terjadinya kanker ? 85% kejadian kanker disebabkan oleh faktor dari luar tubuh: Karsinogen (Polusi makanan /Minuman/ udara/air), sinar UV, virus, infeksi Hanya 15% disebabkan oleh keturunan Bagaimana terjadinya kanker ? Gen Keturunan: 15%i Gen Gen Polusi udara, pencemaran makanan, Uvsinar matahari,, virus, infeks: 85% Sel & Gen Normal Gen Gen Sel & gen Tidak normal Gen Gen Gen Polusi udara, pencemaran makanan, UV/sinar matahari,,kegemukan, hormon Bagaimana terjadinya kanker ? Gen Gen Gen Sel tidak normal,gen tidak normal, menghasilkan jaringan kanker/tumor Polusi: makanan minuman udara, air, sinar, kegemukan, hormon METASTASE MENYEBAR Gen Gen pembetul Normal DNA Gen Gen Kerusakan gen lanjut Gen Diperngaruhi oleh: • Zat gizi Diperngaruhi oleh: • Aktivitas fisik, kegemukan, makanan yang dikonsumsi • Hormon dan faktor pertumbuhan tertentu spt karotenoid, retinol • Serat, bakteri Gen kolon, asam lemak yang mudah Sel normal menguap, pre/probiotik Gen Sel tidak normal Gen Sel bunuh diri Pangan Zat gizi: protein,lemak, karbohidrat, vitamin, mineral, serat Tanaman obat Sumber utama ---------------- Komponen fungsional/bioaktif Non-gizi Kadar rendah Rasa, textur, volume, Penting Kemungkinan toksik Sangat kecil Kadar tinggi tidak penting Besar “Functionality” berdasarkan komponen: Komponen Jenis Fungsi Sumber Zat-zat gizi makro Protein, lemak, karbihidrat Zat pembangun, Energi, pelindung Esensial Biji-bijian, Kacangkacangan, Daging, ikan, dll Vitamin dan Mineral esensial A, B, C, D, E, K, Folat, pantotenat, Niasin, biotin Metabolisme Seluler normal Esensial Sayuran dan buah-buahan, Rumput laut, sintetik Serat Selulosa, pektin, hemiselulosa Gum, oligoskarida Prebiotik, Kontrol kolesterol, Pencernaan,ImunitasDia betes, kanker, kegemukan Sayuran dan buah-buahan, Rumput laut Sintetik “Functionality” berdasarkan komponen: Komponen Gula alkohol Asam amino, Jenis Fungsi Sumber Eritritol Arabitol, ribitol, xilitol Sorbitol, manitol, Sebaian prekursor glikogen, antiketogenik, substitusi gula rendah kalori, kambah, laxative, Anti caries, anti tumor Ganggang, jamur, Exudat tanaman, molases, rumput laut Arginin Aspartat/gin Antihipertensi Fatig kronik, sirosis hati Anti epilepsis Anti insomnia Analgesik Anti depresi/ Parkinson’s diseases Anti depresi/hiperaktif Protein Glutamat Triptofan Tirosin Fenilalanin “Functionality” berdasarkan komponen: Komponen Jenis Fungsi Sumber Peptida Casomorphin Imunopeptida Caseinophosphopepti da Peptida bioaktif Anti diarea Stimulasi imunitas Absorpsi Ca Antihipertensi Kasein susu Bakteri asam laktat Lactococcus, Lactobacillus, Bifidobacterium,dll Lactose intolerant Probiotik: diarea, anti kolesterol/ kanker/ konstipasi imunostimulan, Produk fermentasi Susu, sayuran, dll PUFA, w-6, w -3 LA, LNA, DHA Metabolisme arakhidonat, anti penyakit kronis Lemak tanamn Daun, biji-bijian, ikan. Hidrolisa protein kacang-kacangan, ikan “Functionality” berdasarkan komponen: Komponen Jenis Fungsi Sumber Thioallyl CH2=CH-CH2X X=struktur organik Hypolipidemic Antitrombotik Anti kanker Bawang putih Protese inhibitor Kunitz Anti kanker Kedele, kacangkacangan Chlorophyllins Khlorofol tanaman Antikanker Khlorofol tanaman Lignans Antikanker Estrogen Kedle, gandum PEITC (Phenethyl isothiocyanate) Antikanker Cruciferous Antikanker Kunyit Curcumin I, III Diferuroilmetan “Functionality” berdasarkan komponen: Komponen Fungsi Sumber Karotenoid Anti penyakit degeneratif Antioxidan Sayuran, buahbuahan, teh Gingerols, shogaol Antioxidan, anti ateroskelosis, Pencernaan, Anti kanker Jahe Ubiquinone s, ubiquinols Antioxidan Imunomodulator (AIDS) Minyak jagung, kacang-kacangan Antioxidan, anti kanker Tanaman Flavonoids Fenol sederhana Jenis Quercetin, galangin,Ruti n, diosmin katekin Khlorogenat,e Antioxidan, anti kanker lagat,protokat ecuat, ferulat Teh Tanaman “Functionality” berdasarkan komponen: Komponen Jenis Fungsi Sumber Isotiosianat sulfofran antikanker brokoli Actoxikavikol asetat Fenil propanoid Anti kanker Languas galanga Aurapten (AURA) D-limonen Anti kanker Sitrus Resveratrol Trihidroxistilben Anti kanker Anggur merah Laktoferin Protein Anti kanker Susu Fitosterol B-sitosterol, kampesterol Anti kanker Hipokholesterol Sayruan, bijibijian Saponin Glikosida Anti kanker Kedele Fitoestrogen, lignan isoflavon Antioxidan, Anti kanker Kedele, sorgum, Momordisin Anti kanker paria Cucurbitasin Anti cacing Labu Penggunaan komoditi tanaman pangan sebagai bahan obat (POM) Budidaya: Temulawak Jahe Lengkuas Kencur Cengkeh Kunyit Bengle Pare Tan hutan: Alang-alang Lempuyagn wangi Curcuma xanthoriza Zingiber officinale Roxb Languas galanga (L) Stuntz Kaempferia galanga L Eugenia aromatica L. O.K. Curcuma domestica Val Zingiber purpureum Roxb Momordica charantina L 1554351 1364270 1256148 615418 556500 488686 215432 140427 Imperata cylindrica (L) Beauv Zingiberis aromaticum Vahl 528845 872642 Potensi pangan tradisional sebagai pangan fungsional Bahan baku:prinsipal/suplemen: sayuran buah-buahan, rempah-rempah, ganggang, rumput laut, mikroorganisme, jamur Diet/menu: gado-gado, asinan, rujak, kredok, bubur manado, sayur asam, lodeh, sayur tumis, pindang ikan, sop buntut/kaki ayam, bubur kacang, cincau, wedang jahe, bir pletok, manisan buah, selai Perlu standardisasi bahan/pengolahan dan uji khasiat Ginger (Zingiber officinale Roscoe) extracts increase in vivo human LDL resistance to oxidation and prevent in vitro cholesterol accumulation in mouse macrophage Fransiska Rungkat-Zakaria(1, Aisyah T. Septiana(2 and Sulistiyani(3. 1) Dept Food Technology and Human Nutition, Bogor Agricultural University, Indonesia 2) University of Jend. Soedirman, Indonesia 3) Faculty of Math & Natural Sciece, Bogor Agricultural University, Indonesia Ginger (Zingiber officinale Roscoe) rhizoma INTRODUCTION GINGER Zingiber family The rhizome is commonly accepted as a source of flavor : cooking, drinks, baking Grows well in tropical rainforest climate Traditional beliefs: • prevention of common cold, physiological and stomach disorders, inflammation, diarrhea, etc (Tang and Eisenbrand, 1992) Scientific Reports Antioxidant capacity of ginger extract (oleoresin fractions) in linoleic acid system > a-tocopherol (Kikuzaki and Nakatani,1993) • Oleoresin fractions gingerol, shogaol Suppress 5-lipoxygenase or prostaglandin synthetase (Flynn and Rafferty, 1986) Protect TPA induced ear edema, epidermal ornithine decarboxylase activity, skin tumor promotion in female ICR mice (Park et al, 1998) Protect lymphocytes and hybridoma cells from induced oxidative stress, reduced intracellular free radicals, increase natural killer lysing function toward target cancer cells (Zakaria et al, 1999) Increase Ca++ATPase activities in cardiac sarcoplasmic reticulum (Antipenko, et al.1999) Modified LDL ( Ox – LDL, Ac LDL) Scavenger Reseptor Phagocytosis Endosome UC EC HDL Pinocitosis Translocation UC LDL LDL Reseptor Lysosome Cholesterol up take receptor synthesis Acetate Cholesterol Cholesterol processing Macrophage cholesterol Metabolism Cholesterol exclussion Monocyte Lipid streaks LDL Adhesion entry Lipid oxidation Foam cells differentiation ROS Protein modification Macrophage Uptake LDL Initiation mechanism of atherosclerosis RESEARCH OBJECTIVES • To find out whether drinking ginger water extract can 1. Lower plasma cholesterols 2. Protect plasma LDL against oxidation • To find out whether the protection of LDL can prevent further accumulation of cholesterol in macrophage METHODOLGY Subjects: 24 healthy male students living in the same religious dormitory two groups: treated group and control/placebo group Ginger drinks: • water extract, pasteurized 850C, added with sugar syrup • standardized acceptable drink • given every afternoon at 17:00 • placebo group received syrup without ginger extract Plasma analysis: 1. Total cholesterol (CHOD-PAP method, Boehringer-Mannheim, kit) 2. LDL-C (PVS method, Boehringer-Mannheim, kit) 3. HDL-C (phosphotungstic acid) LDL isolation: • 8 ml plasma + 5 ml NaCL 0.9% + EDTA 0.01% • incubation in polyalomer tubes •Ultracentrifugation at 36.000 rpm 40C, 20 hours • Tube cutting b VLDL LDL + LDL (d> 1.006 g/ml) • Scaled tubes + 0.1109 g KBr/ml • Mixing • Transfer to new polyaomer tubes + 4 ml KBr in NaCl-EDTA • Ultracentrifuse, 36.000 40C 24 hrs • LDL (upperpart, d , 1.063 g/ml) • LDL, dialized LDL oxidation: • The isolated LDL was oxidized with 5 mM CuSO4 at 37 C • conjugated dienes was monitor continuously at 234 nm for 30 minutes 0 MDA LDL analysis: • Extraction with butanol • measurement at 515 nm exitation, 553 nm emission (Conti et al 1991) Cholesterol accumulation in macrophages: • Plasma from one healthy male was incubated with 430 mg/ml ginger dichloromethane extract, 1 mM a-tocopherol or none for 3 hrs at 370C •Mouse was injected with thioglycolate • macrophages, 2-3x106, isolated from peritoneal, were incubated in RPMI 1640 containing 1% BSA and incubated for 4 hrs at 370C, 5% CO2 • cells were washed and cholesterol was extracted with isopropanol • total cholesterol, esterified cholesterol, free cholesterol analysis was done by HPLC RESULTS 180 160 140 mg/dl 120 HDL-c LDL-c 100 Total-c 80 TG 60 40 TG 20 Total-c 0 LDL-c Bef or e TREATMENT Af t er HDL-c Bef or e PLACEBO Af t er Figure 1. HDL, LDL, and total cholesterol, and triglyceride in plasma of male student subjects before and after treatment with ginger drink for 30 days (n=12) and those in control placebo subjects (n=12) nm ol conjugated diena/m g LDL protein 800 700 600 0 t ime of cont rol 500 30 t ime of cont rol 400 0 t ime of t reat ment 300 30 t ime of t reat ment 200 100 0 0 25 50 75 100 125 Tim e, m in Figure 2. Significant protective effects of drinking ginger for 30 days in male student subjects (n=12) on resistance of the isolated LDL against oxidation by CuSO4 for 100 minute, measured by conjugated dienes as the oxidation product during. Figure 3. Cholesterol profiles in macrophages incubated with oxidized human LDL that have been supplemented with ginger dichloromethane extract (430 mg/ml), a-tocopherol (1 mM) or unsupplemented (control) CONCLUSSION 1. Drinking ginger water extract for 30 days did not lower blood cholesterol levels nor reduce plasma MDA 2. However there was significant resistance to oxidation of the LDL against induced oxidative stress 3. The resistance of LDL to oxidation prevent cholesterol accumulation in macrophage cells similar to a-tocopherol IMMUNO ENHANCER ACTIVITY OF GINGER (Zingiber officinale Roscoe) WATER EXTRACT IN HEALTHY MALE STUDENT SUBJECTS Fransiska Rungkat-Zakaria1, Nurrahman2, Dondin Sayuthi3, Francine Belleville4, and Pierre Nabet4 1. Dept Food Technology & Human Nutr, Bogor Agricultural University, Indonesia 2. School of Nutrition, Semarang, Indonesia 3. Deparment of Chemistry, Bogor Agricultural University, Indonesia 4. School of Medicine, Henri POINCARE University, Nancy, France Table. Average and range values of lymphocyte cell analysis of proliferation activities (SI) in the presence of Con A, LPS or paraquat; of MDA cell; and of percentage of CD3 cells Average values No Parameters Subjects 1 SI of cells cultured with Con A Treated group 2 3 4 5 SI of cells cultured with LPS SI of cells cultured with by paraquat MDA cells (mmol/l) Percentage of CD3+ cells Day 0 Day 30 3.33 (2.63-6.40) 7.87 (4.58-10.75) 0.63 (0.46-0.75) 10.42 (4.63-16.75) Treated group 1.39 (1.09-1.80) 2.07 (0.83-3.34) Control group 1.74 (0.98-3.06) 2.05 (0.64-6.08) Treated group 1.15 (0.75-1.83) 2.15 (0.83-5.44) Control group 1.28 (0.79-2.32) 1.62 (0.62-3.94) Treated group 3.33 (2.63-6.40) 0.63 (0.46-0.75) Control group 3.03 (2.55-4.40) 0.83 (0.67-1.06) Treated group 82.42 (77.74-91.29) 80.19 68.15-91-99) Control group 79.52 (63.75-93.15) 78.44 (69.38-87-62) Control group Lysing activity (%) A 35 30 25 20 15 10 5 0 100;1 50;1 B Lysing activity (%) Control group (n=12) Treated group (n=12) 60 50 40 30 20 10 0 100;1 50;1 Control group (n=12) Treated group (n=12) Figure 1.Increasing NK lysing activity of 12 healthy male subjects given ginger drink..Blood was withdrawn for lymphocyte isolation at before treatment (A) and after treatment (B) from control and treated groups. Ginger drink was given for 30 days every afternoon and the cells were incubated with target cells which had been treated with H3-thymidine with the ratio of 100:1 or 50:1. Data was expressed as count per minute of the unlysed target cells. 25 20 15 10 5 0 0 30 Days of treatm ent MDA of treated subjects (umol/L) MDA of control subjects (umol/L) Vitamin E of treated subjects (umol/L) Vitamin E of control subjects (umol/L) Figure 2. MDA and vitamin E content in plasma of subjects before (0 day) and after receiving ginger drink for 30 days (n=12). The subjects were healthy male students living in the same dormitory. MDA was analyzed using the method of Conti et al (1991).Vitamin E was analyzed using spectrofluorometer. A B 1.97 2.09 3.17 3.97 4.18 5.89 6.18 7.90 8.45 10.23 Retention time (min) Figure. Retention time (min) C HPLC chromatogram of ginger oleoresin (A), of plasma from control subjects (B) and of plasma from subjects received ginger drinks for 30 days (C). All samples were extracted with ethanol and injected in Lichosphere 5m OD2 column. Retention time (min) CONCLUSSION • In human study, using male adult healthy subjects, there is improvement in T cells proliferation activities and percentage. • Improvement on T cells was accompanied by resistance of lymphocyte cells against oxidative stress. This improvement was demonstrated by the cells obtained from the subjects drinking ginger • Resistance of the lymphocytes was not accompanied by decrease in MDA cell • Beside the role of ginger compounds as protectant against oxidative stress, they appeared to have other immunoenhancement activity, notably in improving NK cell activity. This improvement in NK cell activity had been observed in the male student studied receiving ginger drink for 30 days • analysis of ginger substances in the plasma reveal the presence of the assumed gingerol group in the plasma of subjects drinking ginger, higher than that of the control group. • Since NK cells are known to have specific activity in lysing mutated or infected cells, the results of this research has shown the scientific support of the traditional beliefs that ginger can improve body resistance to common cold. LITERATURE REVIEW 1. Tang W and Eisenbrand G. 1992. Chinese Drugs of Plant Origin: Chemistry, Pharmacology and Use in Traditional and Modern Medicine. Springer-Verlag, New York. 2. Nakatani N. 1997. Natural antioxidants from spices. In: Huang M, Ho C, Lee CY (Eds). Phenolic Compounds in Food and Their Effects on health II. Am Chem Soc., Washington DC. 3. Hikino H, Kiso Y, Kato N, Hamada Y, Shioiri T, Aiyama R, Itokawa H, Kiuchi F and Sankawa U. 1985. Antihepatotoxic actions of gingerols and diarylheptanoids. J Ethnopharmacol 14: 31-39 4. Kikuzaki H. and Nakatani N. 1993. Antioxidant effects of some ginger constituents. J Food Sci, 58: 1407-1410 5. Zakaria-Rungkat, F, Darsana L and Wijaya H. 1996.Immunity enhancement and cell protection activity of ginger buds and fresh ginger on mouse spleen lymphocytes. Symp Non-Nutritive Health Factors for Future Foods. Korean Soc. Food Sci and Technol, Korea 6. Antipenko, AY, Spielman AI, and Kirchberger MA.1999. Interactions of 6-Gingerol and Ellagic Acid with the Cardiac Sarcoplasmic Reticulum Ca2+-ATPase. J Pharmacol Exp Therapeutics, 290, 227-234 7. Conti M, Morand PC, Levillain P, and Lemonnier A. 1991. Improved fluorometric determination of malonaldehyde. J Clin Chem, 37/7: 1273-1275 8. Fritz KL, Nelson TL, Ruiz-Velasco V, and Mercurio SD. 1994. Acute intramuscular injection of oils or oleic acid component protects mice against paraquat lethality. J Nutr. 194: 425-42 9. Meydani SN, Wu D. Santos MS and Hayek MG. 1995. Antioxidants and immune response in aged persons: Overview of present evidence. Am J Clin Nutr. 62: 146S-147S 10. Zakaria-Rungkat, F., Nurahman, Prangdimurti, E., Tejasari. 2003. Antioxidant and Immunoenhancement Activities of Ginger (Zingiber officinale Roscoe) Extracts and Compounds in In Vitro and In Vivo Mouse and Human System. Nutraceuticals and Foods.8; 96-104 ACKNOWLEDGEMENT This reseach was funded by the University Research Graduate Education)-Project III ’97-2000, Department of Higher Education, Ministry of National Education HEALTHY DESSERT MAKANAN PENCUCI MULUT SEHAT MAKANAN PENCUCI MULUT FUNGSIONAL PANGAN (PENCUCI MULUT) FUNGSIONAL AKTIFITAS ANTI KANKER GEL CINCAU HIJAU (Cyclea barbata L.Miers) Fransiska Rungkat-Zakaria, Endang Prangdimurti, Edna Ananta, Albertus Seno Pandoyo PENDAHULUAN Secara tradisional, tanaman cincau hijau (Cyclea barbata L. Miers) digunakan sebagai obat penurun panas, obat radang lambung, mual, dan penurun tekanan darah tinggi. Minuman cincau merupakan produk olahan yang berbentuk gel dan dibuat dari daun cincau hijau melalui proses extraksi dingin. Beberapa laporan hasil penelitian menunjukkan bahwa tanaman cincau mempunyai aktivitas sitotoksik dan antimalaria (Guinaudeau, 1993). Aktivitas sitotoksik ini dihipotesiskan dapat menghambat proliferasi sel kanker atau sel tumor di dalam tubuh.. Informasi ilmiah mengenai khasiat gel cincau hijau akan mendukung perkembangan produk pangan fungsional yang berbentuk “healthy dessert”, yang akhir-akhir ini amat digemari karena dapat mengimbangi konsumsi diet mengandung lemak dan protein hewani tinggi dan kadar serat rendah. Kanker merupakan penyakit yang diawali dari mutasi gen seluler, yang diketahui sebagian besar disebabkan oleh faktor external seperti polutan kimia baik pada makanan maupun lingkungan, virus, dan radiasi. Mekanisme pencegahan mutasi gen dapat terjadi melalui aktifitas antioxidan, perbaikan imunitas, atau sitotoksik (WCRF&AICR, 1997; Zakaria-Rungkat et al., 2001) METODA PENELITIAN Extraksi air daun segar dengan aquades dilakukan secara manual karena terbentuk gel yang kemudian didinginkan untuk memperoleh cairan sineresis. Extrak air daun, akar dan batang dikeringbekukan. Extrak etanol dan hexan dilakukan pada daun, akar dan batang keringbeku. Extrak di tambahkan pada media kultur sel-sel kanker K562 (turunan sel leukimia) dan Hela (turunan sel kanker servix) dan pada kultur sel limfosit yang diisolasi dari darah tepi seorang mahasiswa sehat, dengan 5 tingkat konsentrasi: C1 (1/2 C2); C2 (setara dengan konsumsi segelas minuman cincau), C3 (2xC2); C4 (4xC2); C5 (8xC2). % Penghambatan 100 50 0 c1 c2 c3 -50 c4 c5 Aid etd hed aib etb heb aia eta hea Konsentrasi Gambar 1. Efek sitotoksik extrak air (ai), etanol (et) dan hexan (he) dari daun (d), batang (b) dan akar (a) tanaman cincau hijau terhadap pertumbuhan sel leukemia (K562). Extrak air akar mempunyai persentase penghambatan diatas 61%, air batang diatas 30%, sedang air daun diatas 37% setelah konsentrasi diatas C3. Peningkatan konsentrasi > C3 (2x 2 gelas muniman gel cincau) tidak nyata menaikkan aktifitas sitotoksik. 50 % Penghambatan 40 30 20 10 0 -10 c1 c2 c3 c4 c5 Aid etd hed aib etb heb aia eta hea -20 -30 Konsentrasi Gambar 2. Aktifitas sitotoksik extrak air (ai), etanol (et) dan hexan (he) dari daun (d), batang (b) dan akar (a) tanaman cincau hijau terhadap kehidupan sel kanker servix (Hela).Ekstrak Aia hingga konsentrasi C4 (setara 4 gelas minuman gel cincau) menghambat pertumbuhan sebesar 31%, extrak aid 22% pada konsentrasi C4. Tabel 1. Aktifitas sitotoksik (persen) extrak gel cincau pada sel limfosit manusia yang dikultur dalam media RPMI 1640 lengkap. Sel hidup dianalisa dengan MTT Sampel Aid C1 C2 C3 C4 C5 -9 -2 -17 -18 -1 Etd -10 0 1 -26 * 7 Hed 49 * 16 8 31* 38 * Aib -11 5 -19 * -39 * -3 Etb -12 -2 -7 -25 * 14 Heb -22 * -11 -2 -7 9 Aia 3 -1 -17 -38 * -52 * Eta -9 11 2 -26 * 10 Hea 31 -14 6 -84 7 (*) menunjukkan perbedaan nyata terhadap kontrol pada selang kepercayaan 95% Bilangan negatif menunjukkan penurunan nilai absorbansi. Tabel 2. Kadar alkaloid extrak cincau. Analisa dilakukan dengan HPLC dengan menggunakan asam scopolamin-HBr dan tropic sebagai standard. Sampel Daun Bata ng Akar Pelar ut Air Etanol Hexan Air *) Scopolamin % b.b % b.k 0.12 0.37 0.69 1.84 *) Tropic acid % b.b % b.k 1.28 3.85 0.16 0.49 - Etanol - - 0.44 1.18 Hexan Air Etanol Hexan 0.07 0.30 - 0.18 0.61 - 1.23 0.59 2.52 1.20 KESIMPULAN 1 2 3 4 5 Minuman tradisional gel cincau hijau merupakan pangan fungsional yang dapat mencegah terjadinya penyakit kanker Extrak yang paling bersifat sitotoksik adalah extrak air akar pada sel K562 (leukimia) yang menghambat kehidupan sel sampai 61% dan pada sel Hela (kanker servix) sampai 31% Rasa extrak akar yang amat pahit dapat merusak cita rasa minuman gel cincau, sedang nilai ekonomisnya amat tinggi Secara keseluruhan semua extrak pada konsentrasi setara dengan satu gelas minuman gel cincau bersifat sitotoksik pada sel K562 dan Hela. tetapi Semua extrak pada konsentrasi setara dengan satu gelas minuman gel cincau tidak bersifat sitotoksik pada sel limfosit normal manusia Ucapan Terimakasih Ucapan terimakasih disampaikan pada Poyek QUE-TPG IPB yang telah mendanai penelitian ini dengan anggaran tahun 2000-2001 Improvement of lymphocyte activity and liver antioxidant in tumour bearing mice after feeding with green gel leaf (Cyclea barbata L Miers) powder Fransiska R. Zakaria1, Sri Yadial Chalid2, Rosanti Setiawati1, Budi Agus Pranoto2, Puspita Eka Wuyung3, and Kusmardi3 1,2) Department of Food Technology and Human Nutrition, Bogor Agricultural University, Bogor 16001, Indonesia 3)Anatomy Pathology, Medical School, Indonesia Univ, Jakarta 10430, Indonesia Bogor Agricultural University Campus IPB Darmaga, Bogor 16002 INTRODUCTION Tumour growth: oxidative stress, immunological disorder, malnutrition Potential role for oxidative-induced injury in the cancer process specifically during the promotion stage (Klaunig et al, 1998) Sources of ROS : from inflammatory cells (Cerutti and Trump, 1991) A need in research to find suitable food for cancer patients Green gel leaf Cyclea barbata L.Miers common refreshing drink in South East Asian countries known traditionally to have cooling and anti inflammation effects in the body This gel /extracts of the leaf had been reported to have : cytotoxic activity on cancer cell lines non toxic on normal human lymphocyte cells (Zakaria-Rungkat et al, 2001) In this study, the leaves were processed to produce powder products which included all parts and fibres in the leaves C3H Mice n=5 Fed with powder for 4-wk (12.1g/kg) Tumor transplantation Tumor latent period (days) Killed , Day 57 Liver Tumor volume (Moving pen, Tajima) Tumor Healthy and necrotic Spleen Spleen cells Tumor cells Resistance to H2O2 (MTT) T cell (Rosette) In vitro cytotoxic toward tumor cell LIVER SOD (Adenochrome asay Misra. H.P. GSH-PX (Paglia and Valentine, 1967 CATALASE Colometri, Sinha 1972 MDA (TBA, Buege and Aust) Total Glutathion DTNB, Ellman. 1982 Healthy tumor tissue Necrotic tumor tissue Latent period, days 3.5 3 2.5 2 1.5 1 0.5 0 LP + T T SC Figure. Latent period of tumor growth in mice fed leaf powder then implemented with tumor cells (LP+T) and in mice implemented with tumor (T). There is no difference in the latent period; the leaf did not prevent tumor growth in mice 2.5 2 Cm 3 1.5 1 0.5 0 LP + T T SC Figure. Tumor volume in mice fed leaf powder then implemented with tumor cells (LP+T) and in mice implemented with tumor (T). The leaf powder decrease tumor volume in mice 4 G r a m s 2 0 LP + T Necrose T SC Whole tumor Figure. Whole tumor and necrotic tissue in mice fed leaf powder then implemented with tumor cells (LP+T) and in mice implemented with tumor (T). The leaf powder decrease tumor mass but increase tumor necrotic tissue in mice 50 40 % necrotic 30 tissue 20 10 0 LP + T T SC Figure. Percentage of necrotic tumor tissue in mice fed leaf powder then implemented with tumor cells (LP+T) and in mice implemented with tumor (T). The leaf powder increase the percentage of tumor necrotic tissue in mice T Cell, % 16,2 8,8 7,5 CP T SC Figure. Percentage of T cells by rosette technique in mice fed leaf powder then implemented with tumor cells (CP), implemented with tumor without the leaf (T) and in standard control mice (SC). Feeding leaf powder increased Tcell inspite of the tumor growth. Tumor implementations without the leaf markedly reduced T cell percentage 77 69,3 80 t D u e m a o d r c e l l s , 59,3 70 60 50 40 30 20 % 10 0 SC T LP+T Figure. Cytotoxicity of T cells from mice fed leaf powder then transplanted with tumor cells (LP+ T), with tumor alone and standard diet (SC). Slight increase in cytotoxicity in T and LP+T, indicate T cell recognition of the tumor antigen Absorbancy, 570 nm 4,50 4,00 3,50 3,00 2,50 2,00 1,50 1,00 0,50 0,00 Without H2)2 NC, 3W With H2O2 NC, 4W LP, 3W LP, 4W Figure. Mice lymphocyte resistance to oxidative stress. H2O2 10-3 M did not influence cells' resistance. Prolonged tumor growth from 3 to 4 weeks markedly decrease cells’ resistance to oxidative stress MDA SOD 15 500 400 300 200 100 0 10 5 0 LP + T T SC LP + T T Figure. MDA content and SOD activity in mice fed leaf powder then implemented with tumor cells (LP+T); in mice implemented with tumor (T) and in mice fed standard diet. The leaf powder increase SOD activity in mice but reduce MDA in the liver SC 600 400 200 0 Catalase GSH-PX LP + T T GSH SC Figure. GSH content and antioxidant enzyme activity in mice fed leaf powder then implemented with tumor cells (LP+T); in mice implemented with tumor (T) and in mice fed standard diet. The leaf powder reduce liver catalase activity but did not effect the other enzyme activity in the liver. CONCLUSION 1. There is no difference in the latent period of the tumor growth in mice. The leaf powder decrease tumor volume and tumor mass but increase tumor necrotic tissue up to 3 times 2. Feeding leaf powder increased Tcell inspite of the tumor growth. Slight increase in cytotoxicity in T and LP+T, indicate T cell recognition of the tumor antigen Prolonged tumor growth from 3 to 4 weeks markedly decrease cells’ resistance to oxidative stress 3. The leaf powder increase SOD activity in mice but reduce MDA in the liver Reduce liver catalase activity but did not effect the other enzyme activity in the liver. Yang sudah/sedang kami teliti: • Kayu manis, cacao • Kumis kucing • Kecombrang • Lengkuas, minyak buah merah, Dll Aspek: imunomodulator, anti kanker anti obesity (baru) Conclusion • Potensi komoditi lokal sangat besar untuk produk pangan fungsional • Bahan baku pangan tradisional mengandung komponen fungsional/bioaktif • Ketersediaan bahan berlimpah • Potensi untuk suplemen dari bahan baku pangan, rempah-rempah, tanaman obat • Rekayasa pangan fungsional dari bahan/resep tradisional • Potensi menu diet tradisional Thank Administration Building Bogor agricultural University Indonesia You