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Development of Functional Foods for Radiation Workers

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Development of Functional Foods for Radiation Workers KR0000208 KAERI/RR-2028/99 Development of Radiation Food and Biotechnology Development of Functional Foods for Radiation Workers 7} Please be aware that all of tbe Missing Pages in this document were originally blank pages 2000. 3. 31. <=> 04 bH TT CD td H| 5|| ^ W ^ -Ml o| ^ o| AH ?=f o ot i. WR ^5]^, glucan ^ interleukin-1, GM-CSF f- -77]- ^ ^l A} 1 III. si (1) ].§- 7f^ A (2) 7fl^ol 5:4 ^^ (2) cells)^ (Nitric oxide) ^ ^ Afl 24 4|i^i Apoptosis (1) 71 AJIS. « 2:1 7]^ ^ (2) j: -fi--fi-33: (3) 2. - in - 2:4 2:4 2:4 ^] paeoniflorin ^5^ f-^ :4 4^AS^fEi ^s]^ paeoniflorin^ - Paeoniflorino| #^^°]^-^# iL^l^l ^^g-1- Ames test°114 ^^ - Paeoniflorin<>1 i^^1^^* iL°l^l S^^-# ^^ decursin 4 decursin angelate-^ -§-s] -§-^§ HPLC ^^ a?i Ad^ 3-A]- ^-^^ -H-M^-^l decursin^l- decursinol angelate^ HPLC iv. 1-1. 71". (1) 44)° - iv - (2) ^IS^l 107H1: 6 (3) 71^ (4) >«^1^ ^^ ^^1 14 ^^ 4-f^i^ SRBCi (1) ^^ ^^^^(stromal cells)^ «!)<£ #^^>^lS(bone marrow stem cells)^ 4£(stromal cells)^-S] - v - (2) 5f7]^ ^ ^|#db^| 1, 87M ^-*1^ ^ 50% ^££ ^ (3) td^Aflio] ^xj-^. ^^(Nitric oxide) S^r NO ^^]# ^-£^^1 ^^Hi^l1?}, IFN-rl- S- NO ^^1- -fr^^^i, iNOS ^l^ NGMMi IL-6, LT, iNOS^ mRNA XI X) apoptosis - vi - (l) (2) ^44 24 4^^6ll4 fl^^ sl^ls ^^ ^7} ^j1) o] (3) i^-g- Afl3£^l Apoptosis apoptgsis 1-2. 7}. (1) ^(herb mixture, HM) (2) 2(HM-II) vii - (3) (7\) HM-I (4) (7\) 47}x] I^r 5.5^11, (4) i^-S- >fli^ Apoptosis ^-^r ^^1 HM-I, HM-H ^ HM-IV^^ s^]^o_^ ^ojA^o|i, apOptosis apoptosis ^1 - vin - HM-I, HM-n ^ ^ 4*1 HM-IMH 2(HM-n)# 44 TTS 3:^-^1-i-(herbal immunomodulator mixture) HIM-I , HM-nS. ^i^*}$!4. QT&Q ^-S. 2:^-^^- HIM-I4 HIM-n^] water extract(W), methanol fraction (M), ethanol fraction(E) ^ polysaccharide fraction(P)# ^2: (1) -I-Pi, HIM-n-PiHH fe^ ^1-f^i^ SRBCi ^^4S.(allogenic lymphocyte)i tfl^ ^^ : HIM-I-W ^^ ir ^^- s?-?]5:r$i4. ££^, HIM-I-Pi HIM- n -Pi ^^ ^Hl^r $t -S-(Graft versus host reaction, GVH): HIM-I-Pi^ HIM-II-Pi - ix - HIM-I-W4 1 ^^^li £tM£] ^^ ^^«}-g-(Mixed lymphocyte reaction, MLR): HIM-I4 HIM-H ^ 4 £ia^ J=LO^ n}~f ^3.*"^ ^slfV til^^^r^ ^ ^#°]^1 ^^^li(allogenic lymphocyte)°ll t^^ MLR(mixed lymphocyte reaction)^ HIM-Ii^ <^1 ^^ ^^°] °-}l}2] ^7} JL^I" i^^l, HIM-H^l (4) HIM-H (2) - HIM-I4 71-^- fe^ SL41- iL°lfe ^r^^r HIM-I-Pi4 HIM-n-Pi #^«V^4. ^14, HIM-I4 HIM , HIM-I-R4 HIM-H-W, Po, Pi ^ W+Pi - x - (4) ^ #^1S tiH^^li HIM-I^l 1- 2:4^4. 3. 14, Hirn-is] 3-f<*)lfe £3£°11 ^^1 IL-104 LT(lymphotoxin)^ ^^# ^-£*>51JI, IGIFGnterferon- r v o inducing factor)^ ^ ?i ^^: f^M^lfe ^^-^ M-E]-^4. 4^i, HIM-I^l A GM-CSFS} IL-37} ^^^ojsfji GM-CSF4 ^Sf^^^^cll Him-I GM-CSF°oH] ^^§io] 14a i4t 4^-^^l ^Xl^l^, Him-II ^ fe ^M1?!, GM-CSF4 EPO(erythropoietin)!- ^ A ^-1: ^7f ZL 14, Him-I ^4fe Him-II ^-^^o] EPO(erythropoietin) f ^1^5L^ ^Sf^^]^: #^§}^ ^^.S. 4^4. HIM-H BE^r EPO - xi - Him-II £ Him-I ^ 4Gray . #, 4Gray Him-II 8Gray (3) (71-) HIM-I-W, HIM-H-W ^ HIM-H-M ^-^, ^-*1 HIM-I-Pi4 HIM-n-M ^ HIM-n-Pi (4) Apoptosis ^^: j HIM-I-W, E, Pi4 HIM-E-W, M, Pi - xii - A] (4) HIM-I-W7} 1.67«fl, HIM-I-H71- 1.5*11, HIM-n-W7> 1.24«fl, HIM-H-Pi ^^ir ^7]-7> (1) ^ 2^ -^4. 3:17]^ M, E, P $14. HIM-E ApoptOSis HIM-I-Wi -l, HIM-E-Wi HIM" "Pit" (2) A] 50% kg ^ 2g O]AJ.O. 2 ^ lOg 6lAj-o. 50% ^ kg^ 0.4gi - xni - 4 GC, GC-MS, NMRf- HIM- HIM- 2. ^^l 10 (1) (2) ^- 2.5, 5, 10 kGy 5 kGy -^-M-, 10 kGy - xiv - 4- #*}<$. 2:4 (1) #44 3=4 10 Si- 10 y (2) 4. 3:4 4. 40] Apoptosis^^ 2:4 ^^-^f" ^^-^^^°H4 apoptosis - xv - (3) #4^ 24 benzo( a )pyrene^S. Salmonella typhimurium TA98, TA100 CHO ] ^ -lr ^S^] CHO ^Si^ benzoC a (4) DPPH<^1 cfl^: ^^f^^^^l €^1 ^## ^ (1) Salmonella typhimurium TA98, TA100, TA102 ^ Ames tesHH ^4^(10 kGy) (2) Hfl^^ i£-fi-^l- 45.°J CHO kGy) xvi - (1) £5] 13; paeoniflorin^ HPLC^ll 13C-NMR #^^4 s§f ^-^ paeoniflorin^ retention time0] 7.915, ^^fa§#€^ 10.914 ^^Si1^. lOkGy 2:4 444 «]24 44^]^^ paeoniflorin ^ 4 4.59%4 4.42%7|- 44 i«l-£H 9X9X3- chromatogram^l pattern (2) Paeoniflorin<>1 w# Ames M paeoniflorin ^453-^, 4 5 ^71-44 ^SJ:aL, wisA]- 44 pattern 4*^41- 444 &£ 3-f 44 4 5^4 44^^^ (3) Paeoniflorin^l CHO 4^tiH^o]lA^ 4°i binucleated cells ir°fl ^A 244^4. 44 paeoniflorin (4) ^^5] decursin ^ decursin angelate^l ^-2] ^^ 44^1 -n-Sl^-&<y decursin4 decursin angelatefe SiO2 column chromatography(tolune: ether, Hexane: EtOAc)^ ^5]4$^^, HPLC1- ^sfl signal^ &*1 xvii - (5) ^3 HPLC ^ ^# Decursin^ decursin angelate^ H^f- Al^ol 3.84 4.0°lSiJi ^iM^ 320 nmi^i 2ltfl^^-£l- 4M-M reverse phase column^] *\ fe acetonitrile: °J:3i^^ O^AJ-co iumn (Shim-pack CLOODS(M)HH $}A^, normal phase columni^^- Hexane: EtOAc -§- . Decursin^ 200ug - lOOOug/ml ^H (6) #*VS ^A> ^^^ -n-Jl^^:^! decursin^l- decursinol angelate HPLC ^^ ^^I^f U1^4 ^^^ -B-^Aou@- ^^14^ decursin^ 0.27%^ decursinol angelatefe ^^ V. 7fl - xvin - SUMMARY I. Project Title Development of Functional Foods for Radiation Workers II. Objective and Importance of the Project The radiation damage has been demonstrated since discovery of X-rays. As the diversity of radiation used in medicine, agriculture, industry, biochemical research and military operation increases, the risk from exposure including total body or local organs damage will be elevated. Thus, the protection of individuals against radiation damage is an important issue. Synthetic compounds have been studied for their ability to protect against the adverse effects of ionizing radiation since the original observation of radioprotection. Subsequently, many compounds have been tested for their ability to modify the effect of radiation. Despite some toxic effects, newer compounds are currently under investigation as possible adjuvants in the radiation treatment of cancer. Natural products such as herbal medicines have only recently begun to receive some attention as possible modifiers of the radiation response. Especially, the regulation of immunity and hemopoiesis have might be a important theme in prevention of disease and overcoming radiation damage. The present study was performed to develope non-toxic functional foods with activity of imrnunomodulation, hematopoiesis augmentation and stem cell protection from radiation. The effects of several eatable herbs and Oriental prescriptions were evaluated, and the mixtures of herbs were made to prepare provisional products with higher activity in all respects. On the other hand, it is required by industrial world to apply the irradiation technology for hygienic purpose that is usually performed by chemical preservatives. So the safety of irradiated functional resources was evaluated in respects of biological activity, genetic toxicity and preservation of active components. - xix - III. Scope and Contents of the Project 1. Development of functional foods for modulation of host defence A. Scope and strategy of the project (1) Selection of test materials We selected 9 herbs and 6 decoctions of the Oriental medicinal prescriptions, concerning usefulness of herbs as foods, quantities of the medicinal plants cultured in Korea, and reports on the effects. (2) Strategy for development of provisional products To develope non-toxic functional foods with activity of immunomodulation, hematopoiesis augmentation and stem cell protection from radiation. Several eatable herbs and Oriental prescriptions were screened, and the mixtures of herbs were made to prepare provisional products with higher activity in all respects. 1.1 Screening of food materials (1) Screening of food materials with immunomodulation activity - Screening of the immunomodulation components from herbs - Investigating synergistic effects of immune cell activation by combined treatments - Investigating the effects of the prescriptions on immune cell activation - Investigating the immunomodulation activity in vivo (2) Screenig of food materials with hematopoiesis improvement activity - Isolation and culture of bone marrow stromal cell - Search for food materials and investigating synergistic effects by combined treatments - Analysis of nitric oxide production in macrophages - xx - (3) Screening of food materials for protection of stem cells from radiation - Selection of 6 energy-tonic or blood-building decoction of Oriental medicinal prescriptions as candidate foods - Evaluating their effects on jejunal crypt survival in irradiated mice - Evaluating their effects on endogenous spleen colony formation in irradiated mice - Evaluating their effects on apoptosis of jejunal crypt cells of irradiated mice 1.2 Development of provisional products for modulation of host defence (1) Preparation of herb mixture and investigation of their effects - Preparation of 4 mixtures of herbs which expected to have the higher activity - Evaluation of the immunomodulation activity of the mixtures - Analysis of effects of the mixtures on the hematopoietic system - Evaluation of the effects of the mixtures on the stem cell protection in irradiated mice (2) Selection of effective mixtures as prescriptions and analysis of their effects - Evaluation of immunomodulation activity of the two prescriptions - Analysis of effects of the prescriptions on the hematopoietic system - Determination of effects of the prescriptions on the stem cell protection in irradiated mice (3) Development of the provisional products and acute toxicity test - Preparation of two provisional products with higher activity of immunomodulation, hematopoiesis augmentation and radioprotection - Acute toxicity test of two provisional products xxi - 2.
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