Y Hoc TP. H6 Chi Minh * Phy Ban Tap 19 * S63 * 2015 Nghién ctru Y hoc KHAO SAT TAC DUNG BAO VE TE BAO GAN CUA CU NGHE (Curcuma longa) PHONG NGUA TON THUONG DO ROI LOAN LIPID GAY RA TREN DONG TE BAO HepG2 Dé Thj Hong Tuoi*, Lé Xudn Léc*, Huynh Thi Kim Loan** TOM TAT Mé du: Dé tai nay tiéh hinh "Khio sdt tie dung bio vé t8’bao gan ciia cle Nghé (Curcuma longa) phong ngiea tn thong do r6i loan lipid gay ra trén dong té’ba0 HepG2". Phuong phép: HepG2 nudi ctiy trong moi tring EMEM, 10% FCS, 2 mM L-glutamin, 100 IU/ml penicilin, 100 gil streptomycin. Xie yj t& bao v6i curcuntin (CM), cao ethyl acetat (EN), cao cbr 70% (CN) tie Nigh ting hitm lneong curcumin 2,5; 5; 10 pgiml (ky higu ND1, ND2, ND3) cb/khéng bé’ sung acid béo 2 mM trong 24 gid, Banh gid téc dung bio vf gan phong ngiea sie ike ché'ting treéng 16 bio, tich by lipid, hogi tie 1é° bio, tink trang stress oxy héa, hoat héa apoptosis. K@t qué: Acid béo 2 mM lim gid 36,6% ty If song cia té bio HepG2. Se ite ché' ting treéng duac phuc Ii6i Khi b6 sung CM, CN 6 oi 3 ning d6 vit EN & NDI v6i bj Ié phng ngien 30 - 60%, Acid béo lam ting tich lity lipid trong t8°bio gan 34,5% so si mdu sinh ly; ND2, ND3 clia 3 mau thie phuc loi tinh trang tich lity lipid thiva v6i ty Ié phng ngiea 25 - 80%. Acid béo 2 mM gitim 37,4% long GSH trong #@’bd0 HepG2; ct 3 ring dé ciia CM, EN, CN déu phuc ii wong GSH 06% higu qué tie 370-490%. Tic dung gy hoai tie t8bao cia acid béo 2 ‘mM gitim 100 - 200% Khi b6 sung CM, EN vit CN. Vé qué trinh apoptosis, acid béo 2 mM lim ting 25,4% ADN phi minh so v6i sink ly; 818 may giim 100 - 200% Kh b6'sung CM, EN vit CN 6 3 ritng do sé Két ludn: Cit Nghé ob tic dung phong ngiea tinh trang tn thneong té'bio gan HepG2 do acid béo 2 mM gy 1a theo thie te curcumin t6t hon ono ethyl acetat vi cao cin 70%, Tie dé 6 thé nghién citu thud tie curcumin hoe cit Nghé dé'phong wiht diéu tri bénh gan do ri loan lipid huyét Tit kh6a: cit Nghé, bio vf tbo gan, rOi loan lipid huyét, acid béo, dng tio HepG2 ABSTRACT STUDY ON HEPATOPROTECTIVE EFFECT OF TURMERIC (CURCUMA LONGA) TO PREVENT HEPATOCYTE LESSON INDUCED BY DYSLIPIDEMIA IN HEPG2 CELL LINE Do Thi Hong Tuoi, Le Xuan Loc, Huynh Thi Kim Loan, * Y Hoc TP, Ho Chi Minh * Supplement of Vol. 19 - No 3- 2015:45- 52 Background: The aim of this work is “Study on hepatoprotective effect of turmeric (Curcuma longa) to prevent hepatocyte lesson induced by dyslipidemia in HepG2 cell line”. Methods: HepG2 cells were cultured in EMEM supplemented with 10% FCS, 2 mM L-glutamine, and 100 IUfml penicillin and 100ugiml streptomycin. Cells were treated to curcumin, ethyl acetate or 70% ethanol extracts from turmeric at concentrations corresponding to that of curcumin at 2.5, 5, 10 ginal alone or combined with 2.0 mM fatty acids for 24h. Hepatoprotective effect was evaluated through several assays including cell proliferation, intra-cellula lipid accumulation, necrosis, oxidant stress, and apoptosis actication. Results: Fatty acid 2 mM reduced 36.6% of HepG2 cells viability. Antiproliferative effect was recovered in * Khoa Duge, Dai hoc ¥ Duge Thinh phd H Chi Minh “Phong Arbovirus, Vign Pasteur Thanh phé H& Chi Minh ‘The gid lién lac: TS. DS Thi Hong Tuoi ‘DT 0908683080 Email: hongtuoid99@gmail.com Chuyén D8 Duge 45Nghién ctu Y hoc Y Hoc TP. Hé Chi Minh * Phy Ban Tép 19 * $63 * 2015 ppresence of CM, CN at 3 tested concentrations and EN at NDI with efficiency of 30-60%. Fatty acids increased 34.5% of cellular lipid accumulation; CM, EN, CN at ND2, ND3 attenuated this situation with efficiency of 25- 802x%, 2 mM fatty acids decreased 37.4% GSH content in HepG2 cells; 3 tested concentrations of CM, EN, CN enhanced GSH levels with efficiency of 370-490%. Necrotic effect of 2 mM fatty acid was decreased 100-200% in presence of CM, EN, and CN at 3 tested concentrations. In ferm of apoptosis, 2 mM fatty acids increased 25.4% DNA fragmentation compared to control. This parameter was decreased by in presence of CM, EN, and CN at 3 tested concentrations with efficiency of 100-200%. Conclusions: Turmeric was shown to prevent HepG2 cells injury induced by 2 mM fatty acid: hepatoprotective effect of curcumin was better than that of ethyl acetate and 70% alcohol extracts. Therefor, turmeric andior curcumin could be studied to develop drugs for preventionltreatment of liver diseases induced by dyslipidemia Key words: dyslipidemia, fatty acids, HepG2 cell line, turmeri, hepatoprotective effect. DAT VAN DE RGi loan lipid huyét la bénh ly khé phé bién va o6 xu hudng ngiy cing ting. Sw gia ting néng d6 lipid trong mau vugt qué kha ning thodi héa hay bai xuat ciia gan dn dén tich tu lipid trong gan, chit yéu la triglycerid, gay ra gan nhiém mé. Sy tich lay lipid thira lam tang peroxyd héa 6 gan tao géc ti do, kich thich biéu hign TNF-a, gay t6n thong ty thé va ADN, hoat hhéa qué trinh apoptosis va hoai tir viém té bao gan”, Xu huéng diéu tri hién nay 1A str dung thudc bao vé gan cé nguiin géc thao duge vi cé thé sir dung lau dai va tinh an toan cao. Céc nghién ctu cho thay curcumin trong ci Nghé (Curcuma longa) c6 Kha ning chéng oxy héa manh nén of tée dung logi bd céc géc ty do va céc tée mhan gay bénh gan nhw ethanol, thioacetamid, carbon tetraclorid...08. Tai Viet Nam, céc nghién airu vé kha nang ting dung cit Nghé trong phing va diéu ti bénh gan néi chung cing nhu bénh gan do réi loan lipid huy@t con han ché. M6t trong nhiing céng cu quan trong dé nghién cru sing loc va phat trién céc chai/duuge ligu c6 kha nang lam thudc la m6 hinh in vitro sir dung céc ding té bao véi nhiéu uu diém nhu thoi gian ngan, it ton kém, sir dung, luong miu it, ¢6 thé tign anh véi s6 luong mau Ion... Hign nay, trén thé gidi, m6 hinh in vitro six dung dang té bio gan HepG2 dang duge img dung ngay cing nhiéu trong sang loc cic chat/durgc ligu o6 kha ning lam thuée phong va 46 iu tr bénh gan. Tir nhimg co sé trén, chting t6i thyc hign dé tai “Khao sat tac dung bao vé té bo gan cia cli Ngh@ (Curcumia longa) phdng ngira t6n thuong gan do rd loan lipid gay ra trén ding t& bao HepG2” véi muc dich sang loc téc dung bao vé té bao gan ciia cao cén 70%, cao ethyl acetat va curcumin ciia ci Nghé trén m6 hinh mé phang tinh trang tén thuong t@ bio gan thudng gp trén bénh nhan bj bénh gan do réi loan lipid huyét. VATLIEU-PHUONGPHAPNGHIENCUU Vat ligu Dong tébao Dong té bao HepG2 (ATCC HB8065") hoat héa va nuéi cay tai Vign Pasteur TP. HCM. Héa chat MGi truéng EMEM, huyét thanh bao thai bé (FCS), trypsin-EDTA (Gibco, Mj); L-glutamin, penicilin - streptomycin, albumin huyét thanh bo, dém phosphat, trypan blue, MTT, dau dé O, Brilliant G250, L-lactic dehydrogenase, acid oleic, acid palmitic, quercetin, curcumin (Sigma- Aldrich, Mj), kit LDH (Roche Diagnostic, Ditc), diphenylamine (Merck). Tat ci héa chat dat tiéu chuain tinh khiét cip d6 1. Acid béo 2 mM duge chuan bi ngay trude khi xir ly t€ bao tir dung dich me 100 mM trong methanol gém acid oleic acid palmitic (ty 1é 2:1) pha loang 50 Tan véi méi truing chia 5% albumin huyét thanh ci, vortex, loc qua mang loc 0,22 ym. Chuyén D8 DugeY Hoc TP. H6 Chi Minh * Phy Ban Tap 19 * S63 * 2015 Mau thit Cao cn 70% (CN) Chiét nga kiét voi cn 70% tir cit Nghe (Krong Béng, Dak Lak), # 1é 1:10. Ham Iugng, curcumin trong cao dinh lugng bing phuong phap UV-Vis 18 10,65% (kW/kl). Cao ethyl acetat (EN) Ha cao con 70% voi nudc, chiét léng-long véi ethyl acetat, thu dich ethyl acetat va c6 dudi 4p suat gidm. Ham Iuong curcumin dinh luong bing UV-Vis la 30,45% (KI/K)) Mau CN, EN dat tiéu chuan co sé va duoc cung cp béi Trung tim Sim & Duge ligu Tp. HCM. Curcumin (CM) Hén hop gém curcumin I (75%), curcumin I 0. Trong tht: nghiém khdo sat hoat tinh dc t@ bao cla 3 mau thir & cdc néng dé tuong ting véi ham lugng curcumin tir 0 d&n 40 g/ml cho thay cdc niéng 46 nhé hon 10 jig/ml khong dc té bio nén duge chon dé khao sét téc dung bio vé té bao gan khéi céc tén thuong do réi loan lipid huyét. Bang 1: Nong df khéo sét ctia ote méu thie Caoethyl | Cao cin 70% acetat (EN) (cn) Ham ivong curcumin 3045 70.65 (klk) rong cao 1D Am cia cao (%) 1342 Tar Néng 4 cudi cing trong mai tivong nudi cay (yal) ‘urcumin (CM)] Caoethyl | Cao cén 70% acetat (EN) (en) noi [25(emy [ 95iens) | 27.2(0nt) Nez [_5.0(CM2) [_19.0(EN2)_[_54,4(CN2) NO3 [10.0 (CM3)_|_37.9(ENS)_| 108.7 (CN3) Nuéi cdy va xir ly té bao T& bao HepG2 duge nui cdy trong méi trudng EMEM, bé sung 10% FCS, 2 mM L: glutamin, 100 IU/ml penicilin, 100 ug/ml streptomycin. Té bao durge i & 37°C, 5% CO: dén, khi d@ phi 70 - 80%, Thu té bio theo quy trinh xtt ly v6i trypsin, dém s6 té bao séing (khéng bat mau xanh) bang trypan blue, cay chuyén hog Chuyén D8 Duge Nghién ctu Y hoc chia vao dia nudi cdy voi mat d6 2,5 x 10¥/em?, U 24 gid 6 37°C, 5% COx, xit ly té bao vai CM, EN va CN tuong timg him lugng curcumin 2,5; 5; 10 ig/ml (ky hiéu: ND1, ND2, NDB) hoc quercetin 10 uM cé hoa khéng bé sung hén hgp acid béo 2mM trong 24 gid. anh gid tilé té bao sdng bang test MIT Ty 18 48 bao sding duge xdc dinh qua hoat ‘tinh enzym succinat dehydrogenase (SDH) cia ty thé chi c6 trong té bao séng. SDH chuyén MIT. [3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromid)] thanh tinh thé formazan tan trong isopropanol tao dung dich mau tim, OD do 6 570 nm phan Anh sé lugng té bio sng 66 trong mau. Danh gid tinh trang tich ly lipid bing phuong phip nhugm dau dé O Dau dé O [a thuée nhudm lipid, chi hoa tan trong lipid. Chat tao mau lysocrom di chuyén tir dung méi vao céc hat mo trong té bao gan lam ching c6 mau dé quan sit duge dui kinh hign vi quang hoc. Céc hat mo sau khi nhuém mau véi dau dé O duge hda tan trong isopropanol tao dung dich mau dd. Gid tri OD & bude séng 500 nm phan anh Iuong lipid trong té bao & mdi giéng. Dinh lwong glutathion (GSH) ngi bao Su ting sinh céc gc oxy héa lam giam GSH ngi bao. Dich ly gidi t& bao dugc bé sung acid tricloroacetic 10%. Ly tam lay dich trong, thém dgm EDTA-phosphat, ii véi thuée thir Ellman [5,5-dithiobis-(2-nitrobenzoic acid)] 6 nhigt d6 phing trong 5 phitt. Do OD 6 412 nm. Ham luong GSH dugc tinh dua theo dudng cong chudin d6 pha tir GSH chudin va thé hién dudi dang mM/ug protein. Trong dé, protein ton phan cia dich t@ bao duge xdc dinh bang phuong phdp Bradford véi thuéc nhuém ‘Coomassie va gam mau chuan la albumin huyét thank ciru (0-4 mg/ml). Dénh gid tinh trang hoai tir té bao gan Tinh trang hoai tt duge danh gid thong qua 47Nghién citu Y hoe hoat tinh aia enzym néi- bao lactat dehydrogenase (LDH) phéng thich vo moi trudng nuéi cay bang kit "Cytotoxicity Detection LDH’, LDH xtic tée phn img kép chuyén pyruvat thanh lactat, tetrazolium mau ving thanh formazan mau dé, do OD 6 492 nm. Xéc dinh hoat tinh LDH dua vio duimg cong chun 46 (0-1000 mU/ml) cua LDH chuain. Dénh gid qué trinh apoptosis Quan sét t€ bao apoptosis duéi kinh hién vi hujnh quang sau khi nhuém véi hén hop acridin orange va ethidium bromid, ADN phan manh duge dinh hong bing thuée thir diphenylamin tao dung dich cé mau duge do OD 6 600 nm va quan sdt bing k¥ thuét dién di trén gel agarose 2%. Higu qua phong ngira tén thiong té bio gan Tac dung bio vé té bio gan danh gid qua higu qua phong ngita tén thuong do acid béo theo céng thie: HQ (%) = (Két quasi so - két Udita os gas) (Kt qUudsss so ~ KBt quanta ¥) x 100 (1) hoe HQ (1) = (Két quansturrémncasavio - kt quaest s)M(K8t quinsssnnsy- Két quadssavis) x 100 (2). Ap dung céng thitec (1) vai tinh trang tich lay lipid, hoai ti t bao, ADN phan mink; céng thite (2) véi test MIT vi GSH. Phan tich két qua va xtrly théng, Y Hoc TP. Hé Chi Minh * Phy Ban Tap 19 * S63 * 2015 M&i thi nghiém dugc lap lai 2 hod 3 Ian. Két qua trinh bay dudi dang trung binh + sai s@ chun ciia gid tri trung binh (Mean + SEM) cua 6 mau cho mét diéu kién nuéi cay trong mot thi nghigém dai dién va duge danh gid nghia théng ké bing phép kiém Mann- Whitney trén phn mém SPSS 17.0. Su khéc iét c6 ¥ nghia khi p < 0,05. KET QUA VABAN LUAN Tac dung phong ngita sw tre ché ting trugng Su ting trudmg cia té bao HepG2 6 mat dé nudi cay 4.2 x 10* t@ bao/emé, sau 24 gid tigp xe véi hn hop acid béo 2 mM va mau thir duge thé hign qua céc Bang 2. Mic dé ting trudng cia té bio HepG2 duge xit ly voi mau thir riéng lé khéc bigt khong 6 ¥ nghia théng ké so voi mau sinh ly (p > 0,05) ngoai trix ND3 cia CM lam ting ty Ié t@ bao HepG2 séng (p < 0,05), chting té mau thir khéng dc té bao é ning d6 khdo sat. Két qua nay phit hop véi thir nghiém vé déc tinh 8 bio. Acid béo 2. mM Lam gidm 36,6% ty 18 sOng ciia té bao sau 24 gid (p < 0,05). Su ttc ché tang trudng duge phuc héi khi b6 sung CM, EN, CN vio méi trudng (p < 0,05). Ty 18 phong ngtra sw tic che tang trudng té bao HepG2 nam trong khoang 30 - 60% (Bang 2). Baing 2: Tac dung phong ngiea swe téc ché ting trusing t@ bao do r6i loan lipid sau 24 git (ODzr trung binh = SEM ‘Acid béo 2 mM () ‘Acid béo 2 mM (+) Kh Curcumin (GM) O80 ethyl acotat |” Cao cbn 70% | Curcumin | Cao ethytacetat [Cao cin 70% Néng 49 cM. (EN) (cn) (cm) (EN) (cn) Not [014920008 | 014920006 | 1730007 [0,117 +0008] 0,13240,008" | 0,113 0,005 NO2 | 0.167 20,007 | 0,15020,010 | 0,16120,007 |0,12820,008"| 0,097 0,002 | 0,128 x 0,007 NOS [0,1860,0127| 0,152=0,008 | 0,174=0,011 |0,126+0,004"| 0,088+0,005 | 0,112 0,003 ‘Ching (0,183 + 0,008 0,097 = 0,005" ‘Quercetin 02162 0,013" 0,110 0,008 % phong ngira sy Gre ché tang trwéng so vei mau ching ben Not 35.7 22 288 Noz 55.3) 03) 550) Nos BiT, “162 273 ‘Quercetin 23.4 “Sp < 005: 50 08 nu sind 48 p< 0,05 50 0 miu a i acid bo 2 ND Chuyén D8 DugeY Hoc TP. H6 Chi Minh * Phy Ban Tap 19 * S63 * 2015 Tac dung phong ngita su tich lity lipid ni bao Két qua phuong phdp nhudm véi dau dé O danh gid kha ning phong ngita su tich ly Nghién ctu Y hoc lipid trong té bio gan HepG2 do ri loan lipid gay ra sau 24 gidy xt ly bay trong Bang 3. du thir durge trinh Baing 3: Tic dung phong ngiea sw tich lity lipid thiva trong t@’bio gan do ri loan lipid sau 24 gio ‘ODew trung binh + SEM ‘Acid béo 2M () ‘Acid béo 2 mM (# "YHIGU Ls raumin (om] C8 ety acetat | Cao cin70% | Curcumin | Caocthytacetat [Cao cin 70% Néng a6 CM EN) (cn) (cm) EN) (cw) Not 071740033 069520078 | 068020025 | 09750023] 085020020" | 7,043.2 0076" No2 | 0,727+0,021| 0,6700031 | 0,734 + 0,034 [0,894 =0,027"| 0,856 + 0,036" | 0,875 + 0.02 Nps | 0.6990,013| 06830012 | 0,737#0.031 [0,705 =0,013"| 0.780017" | 0,856+0.027 Ching 0,733 +0011 0,986 0,022" Quercatn 0,794: 00117 0,873 + 0,028 Fe phong nga sy Wich ly tnd tha so wi mBu chong BEN NOT 43 302 2s NDZ 36,3 513 438 NDB 756 a3 B19 Quercetin 447 *p<0,05; % p<0,01: so i miu sin by ©: p< 005: so wi mt aia béo 2 mM be sung quercetin 10 uM Ca 3 ning d6 cia 3 mdu thir riéng 1é Khong nh hung dén tinh trang tich ly lipid thira trong té bao gan HepG2 (p > 0,05). Acid béo 2 mM lim ting tich lay lipid trong t@ bao gan 34,5% so véi mau chting sinh ly (p < 0,05); téc dung nay dio ngugc khi bé sung quercetin 10 UM hodc cao/hoat chat tir cl Nghé (p < 0,05). 0 4 3 mau thiz, NBS cho higu qua phong ngiza tét nhat; NB3 cia mu CM, EN han ché tich kay lipid thira t6t hon 06 ¥ nghia théng ké so véi quercetin (p < 0,05). 6 ciing 1 niéng d@ curcumin, cao EN cho téc dung phng ngita tich ly lipid t6t hon CM rdi dén CN, Tac dung phong ngita tinh trang ting sinh gécty do Tac dung chéng oxy héa duge danh gid qua Bang: "p< 005 30 mi ong ben I (ac bo 2 mM) vige lam tang him lugng GSH néi bao. Két qua duge trinh bay trong Bang 4. Sau 24 gid tip xtc, cd 3 ning d6 cia CM, EN, CN va quercetin 10 iM d@u lam giam lugng GSH khéng c6 ¥ nghia théng ké so véi mau sinh ly (p > 0,05). Lugng GSH trong té bao HepG2 tip xtc véi acid béo 2 mM gidm 37,4% so véi mau ching sinh ly (p < 0,05). Sw od mat cia, quercetin 10 iM gitip phyc Koi hrong GSH bi gidm do glucose 20 mM gay ra (p < 0,05). Ca 3 néng d@ khdo sét ctia CM, EN va CN déu thé hign téc dung ching oxy héa: higu qua phuc hoi ham lugng GSH tir 370-490% so véi mau acid béo 2 mM mdt minh (p < 0,05). Tuy nhién téc dung nay cia 3 miu thir thip hon so véi mau d6i chiing quercetin 10 uM (p < 0,05). dung phong ngiea tinh trang stress oxy héa do r6i logn lipid saw 24 git Ham [wong GSH (uM/mg protein) trung binh # SEM. Acid béo 2 mM (-) ‘Acid béo 2 mM (+) KY higuNEng a | Curcumin ] Cao eth acetat | Cao cn 70% | Curcumin | Cao ethyl acetat | Gao cbn 70% cM (em) EN) (en) (em) (EN) (en) NOT wae7a | e2t243 | 812220 [i726 T6299 | 55275 NZ 48,6421 49,4239 582246 156,0 + 3,8" 131,323,08 152,5.9,9°% NDS 60,7 7,2 58,4236 58,725,7, 1547236" | 142,722,6" 135,7 25,6" Chuyén D8 Duge 49Nghién cru Y hoc Y Hoc TP. H6 Chi Minh * Phy Ban Tap 19 * S63 * 2015 Ham lung GSH (uM/mg protein) trung binh t SEM ‘Acid béo 2 mM () ‘Acid béo 2 mM () Ky higu Neng 4 | Curcumin | Cao ethyl acetat | Cao cin 70% | Curcumin | Caoethyl acetat | Cao cdn 70% om (em) (ey) (cn) (em) (EN) (ew) Ching 63,1270 3520 Quercetin 35 722.4 174327, % phong ngla inh rang sress ony ha so voi mBu chang BETA NOI 3716 ag ol NDZ 4954 73887 4789 NOS 4880 437.5 4076 uereatn S711 “p< 005; p<0,01: 0 06 miu sink If *: p< 0,05: 50-06 mw acl bo 2 mb sung quercetin 10 uM Tac dung phong ngira tinh trang hoai tir Tinh trang hoai tir duge danh gid qua hoat tinh enzym lactat dehydrogenase (LDH) phéng thich tir té bao gan vao méi trudmg do mang t& "p< 0,05 50 mu bf i ac bo 2 mM bao bi tén thuong. Tac dung phong ngira tinh trang hoai tr t® bio gan HepG2 cia CM, EN va CN duge trinh bay trong Bang 5. Bang 5: Téc dung phing ngita tinh trang hoai te tébo gan do réi loan lipid sau 24 gid Hoat tinh LDH (mUimi trung binh # SEM Aeld béo 2 mM (>) ‘Acid béo 2 mM (#) Kyhigu Neng a | Curcumin] Cao ethyl acetat | Cao cin 70% | Curcumin | Cao ethyl acetat | Cao cbn 70% cM (cm) (EN) (cn) (cm) (EN) (CN) NOt 775228 | 102th To4eto | i7ete | 95206" | 180221 NOZ esto | 119228 157208 | 1062129 | 112218 1302175 NDS 74209 | 117215" T4215 | 53204 T2227 a5208 Ching 175208 287220" Quercetin 172219 18.9212 "Zh phong nira tinh Wang hoal Wi so voi mau chang bonny NOt 98,0 1703 5A NBZ 1607 155.6 139.1 NBS 207.4 1553 170.4 Quercetin ara Cao EN va CM khi diing riéng 1é lim giam tinh trang hoai tu, giam phéng thich LDH cé y nghia théng ké so véi mau sinh ly (p < 0,05), tt hon quercetin 10 1M. Sau 24 gid’ tip xitc, acid béo 2 mM gay hoai tir bao gan, ting hoat tinh LDH ngoai bao 64,0% so véi mau sinh ly (p < 0,05). Tac dung nay giam khi bé sung mau thir CM, EN va CN 6 ca 3 ning d6 Khao sat (p < 0,08). Da sé cdc ning a6 Khao sat cia CM, EN, CN cho téc dung tét hon quercetin 10 uM (p< 0,05). Tac dung phong ngita tinh trang hoat héa qua trinh apoptosis Trong bénh gan, té bao gan nhay chm véi te 50 nihan/y€a t6 doc t8 bao dan dén hoat héa qué trinh apoptosis gay phan manh ADN. Két qua phong ngiza phan minh ADN dugc trinh bay & Bang 6. Két qua cho thy quercetin va cao EN khi dig rigng 1é lam giém tinh trang phn manh ADN; CM va CN chi lam giam tinh trang nay 6 1 néng 46 (p< 0,05). Sau 24 gid, acid béo 2mM gay hoat héa apoptosis té bao gan, ting 254% ADN phan manh so véi mau sinh If (p < 0,08). Téc dung nay giam khi bé sung mau thir CM, EN va CN 6 ca 3 ning d@ Khao sat (p < 0,05). Da sé néng d6 khao sét cia CM, EN, CN cho téc dung t6t hon quercetin 10 uM (p < 0,05). Két qua nay hoan toin phi hop véi hinh anh quan sat duge Chuyén D8 DugeY Hoc TP. H6 Chi Minh * Phy Ban Tap 19 * S63 * 2015 bing ky thudt dién di ADN phan manh trén gel Nghién ctu Y hoc agarose 2% (Hinh 1). Hinth 1: Két qua dign di trén gel agarose 2% phan tich ADN phan manh Baing 6: Tic dung phomg ngiea tinh trang phitn mank ADN trong t&’bio do réi loan lipid sau 24 gio % ADN phan manh trung binh + SEM ‘Acid béo 2 mM () ‘Acid béo 2 mM (+) Ky higu Neng a} | Curcumin | Cao ethyl acetat | Cao cin 70% | Curcumin | Cao ethylacetat | Cao cin 70% cM (CM) (EN) (cn) (CM) (EN) (CN) NOI 42.8224" 4682 1,1 469216 | 400217 | 389244 49.2210" NDZ 490235 | 432238 491225 | 41626) 25238 52623, NOS 497260 | 418237 46,1220" | 45825; 02s 1 6242088 Ching 540428 67.7420" Quercetin 425514 S22221 ephéng ngira sw phan manh ADN so voi mau ching bénh ly NOt 202.5 2105 135.8 NBZ 1906 184.6 110.6 NBS 161.7 127.9 38.9 Quercetin 1136 “p< 0,05: 50 0 mi sin lg *p<005:s0 i miu acid do 2 mM bi sung quercetin 10 uM Bang 7. “p< 00550 di mu bf i (ac bo 2 mM) Két qua téng hop tic dung bio vé t€ bao gan ciia cic mau thir tir ca Ngh@ duge trinh bay & Baing 7: Mitc dé bio v¢ t@’bio gan ciia CM, EN, CN phing ngita sue ite ché'ting trading (TT), tich lity lipid (LP), ting gde oxy héa (OX), hoai tie (HT), hogt héa apoptosis (AP) do acid béo 2 mM. Nang a6 ‘Curcumin (CM) Cao ethyl acetat EN) Gao cin 70% (EN) onaae [arpup] ox [ut[aP| tr] ue | ox | wt | AP | tr up| ox | aT | AP wort - > [+ lp = pepe ~+)>-p ps [+ noe ets) fel] = De > |=[+)> >=] nos [+[=[|+|e>-]*[~* > ]+)+|+|*]=le Guewein [+ [+ [rele] + ing 06 tic dung ut Trong dé tai, hén hop acid béo 2 mM da gay duge tinh trang tén thuong té bio gan, két qua nay hoan toan phit hop véi bio céo trude day. Tinh trang tén thuong nay gay ra trén t@ bao HepG2 giam khi cé mat quercetin 10 iM, chat da duge chtimg minh cé tac dung bao vé té bao gan®, Két qua khao sat téc dung bio vé té bao gan cua CM, EN, CN cho thay curcumin thé hién Chuyén D8 Duge Hy, M4, Her 26 te dug wai cae mite df Khe nhaw hoat tinh bao vé té bao gan tat hon cao cén 70%; cao ethyl acetat. Su khéc biét gitta cao ethyl acetat va cao cBn 6 cde ning d6 Khao sét Khong, dang ké (p > 0,05). Digu nay hoan toan phi: hyp céng bé curcumin 1a hoat chat chinh trong cu Nghé®. Curcumin 1a mt polyphenol, diéu nay 6 thé giai thich cho tde dung tuong ty gita cao toan phan va cao ethyl acetat. Tir dé goi ¥ sir 51Nghién citu Y hoe dung cao cn toan phian hoc hoat chat curcumin 46 nghién cu phat trig thude ding trong phang vayhoac diéu tri bénh vé gan. Téc dung bao vé t& bao gan chéng lai tn thuong do réi Joan lipid huyét ciia ct Nghé hodn todn phit hp véi céc béo cdo trude dayt+®, That vay, mot sé béo céo vé ché dG an chita céc hgp chat curcuminoid ciing nhw cao chiét tir Ngh@ lam gidm cholesterol, triglycerid huyét thanh va gan chugt duge cho Sn cholesterol. Ngoai ra, curcumin cé kha ning hoat héa AMPK (adenosine 5'-monophosphate-activated protein Kinase), lim gidm biéu hién SREBP-1 (sterol regulatory element binding proteins-1) va FAS (fatty acid synthase), din dén tte ch€ qua trinh téng hop lipid & gan va kha ning chong oxy héa gan qua co ché ting biéu hign cia PPAR (peroxisome proliferator-activated receptor)” KETLUAN Ca Nghé thé hign tac dung phong ngiza su ie ché' ting trudng té bio, tich lay lipid, tinh trang stress oxy héa, hoai tt va apoptosis té bao gan HepG? do acid béo 2 mM gay ra. Téc dung bao vé t@’bio gan cia curcumin tét hon cao ethyl acetat vi cao cn 70%. Két qua cho thay curcumin déng vai trd quyét dinh tac dung bio v6 gan cla ci Nghé; tir dé 6 thé nghién ctu, phat trign thude tir curcumin hoe cu Nghé dé phing vayhodc diéu tri bénh gan do ri loan lipid huyét, TAI LIEU THAM KHAO. 1. AVDosart MS (2014), Red cabbage (Brasica oleracea L.) mediates redoxsensitve amelioration of dyslipidemia and Y Hoc TP. Hé Chi Minh * Phy Ban Tap 19 * S63 * 2015 hepatic injury induced by exogenous cholesterol administration, Am J Chin Me 42() 189-208, 2 Alla M, Mateos R, Ramos 5, TecumberF, Bravo T, Goya 1 (2006), Influence of quercetin and rutin on growth and antioxidant defense system of a human hepatoma cell line (Cepia), Eur | Nut, 450) 19-28, 3. Alia M, Ramos §, Matecs R, Granado-Serrane AB, Bravo L, Goya L. (2006). Quercetin protects human hepatoma Hep? against oxidative tess induced by tert butyl hydroperoxide, Texio! Appl Pharmac, 212 10-118 4. Asai A, Nakagawa, Miyazawa T (1999) Antioxdative fects of turmeric, rosemary and capsicum extracts on membrane Phospholipid peroxidation and liver lipid metabolism in mice, Bic Biotec, ios, 6: 2118-2122 5. Babu PS, Srinivasan K (1997). Hypolipidemic action of ‘curcumin, the active principle of turmese (Cur Fong) in sreptovotocin induced diabetic rats, Molecular end cli ices, 1651-2) 169-175, 6. Babu PS, Stvasan K (1997). Hypolipidemic ection of ‘curcumin, the active principe of turmeric (Cureura longa in Ssteptozotocin induced diabetic rats. Mal. Ci Bacon, 166:169-175. Bone K, MillS @013). Principles and Practice of Phytotherapy ‘Moder Herbal Medicine, Elie Heth Since $04.90, 7. Kang OH, Kim $B, Seo ¥S,Joung DK, Mun SH, Chot JG, Lee "YM, Kang DG, Lee HS, Kwon DY 013). Curcumin decreases oleic acid-induced lipid accumulation via AMPK ‘phosphorylation in hepatocascinoma cells Eur Rep. Ma Pharmac! Si 17(19} 2578-2586 Menon VP, Sudheer AR (2007). Antioxidant and ant- inglammatory properties of curcumin, Aeones in experimental medicine and bly, 595: 108-125, 9. Naik RS, Mujumdar AM, Ghaskadbi $ 2008), Protection of liver cells from ethanol cytotoxicity by curumin in live slice culkin vito, oul of etinopharmacsogy 95(0 31-37 10. Rao DS, Sekhara NC, Satyanarayane MN, Srinivasan M_ (197, Efecto curcumin on serum and tivercolesteral evels Inthe rat Nutr 10007-1316 [Negay nh ba bt: 3ornoro14 [Negy phn bigm nin xét bi bio: 25/11/2014 [Nga bai bo dupe ding: 15/05/2015 52 Chuyén D8 Duge