NGHIEN ctfu DOC TINH CUA 3-MONOCHLOROPROPAN-l,2-DIOL (3-MCPD) TOEN HINH T H A I HONG CAU VA SU" TAO VI NHAN
Ngd Kien Die', Tran Manh Hioi^
TOM TAT
Ma diiu 3-MCPD la mot duyham diwc tae, ra trang ,,ua trnih thiiy phan protein thiec vat dieai sir xuc tic ciu, acid Da CO nhieii nghiin ciru chieng minh doc Hnh am 3-MCPD tren than, ca quan sinh due vi tim nmg gm, ung thir. Trang n^ien cin, niy, ehimg toi tich hinh thea doi doc tinh dm 3-MCPD tren hinh tha, hSng eiu va sie too vi nhdn.
Muc tiiu: Nghien aht ndy bao gom 2 muc tieu sau: 1) Nghien deu dgc tinh dui 3-MCPD tren hlnh thai dm hong cau, va 2) Nghien deu doc tinh cua 3-MCPD tren nhihn sdc tlte'qua phuang phdp quan sdt su hinh thdnh vi nhdn.
Phuang phdp: Chugt nitdt trdng diegc dio uo'ng 3-MCPD trong 6 tlidng a cdc lieu 1 mg/kg, 10 mg/kg va 20 mg/kg. O thiri dian k^ thiic kitdo sdt isau 6 tltdng), mdu diugt thi nghiem diegc thu tliap va khdo sdt hirdt thai Itong cdu vd su too vi nhdn dtra tren phieang phdp nliuom man bang Giemsa
Kei (fud: 3-MCPD gay thay ddi rd ret hinh thai hong cau: gay gidm tlte tidt hong cm, tdng sac tova diuyen sang dgng hong diu gai. Ben canh dd, sir dung keo ddi 3-MCPD cdng gdy sif gia tdng hhdt tlidnh cdc vi nhdn tren hong cdu (mot bieu hien dm ton thieang nhidm sdc the). Nhieng tdc dgng ndy dm 3-MCPD tdng rd lum ldu de ditng kem theo dhanol.
Kii ludm 3-MCPD a cac lieu 1 mg/kg, 10 mg/kg vd 20 mg/kg sir dtmg h-ong 6 thdng gay thay ddi hhth thai hong coil vd gdy ton thuang nhiem sdc the, vi theco the tdc dgng deh diitc ndng hong cau vd nguy ca gay ung thu.
Tit khoa: 3-MCPD, hdng cdu gai, vi nhdn. dgc tinh man tinh ABSTRACT
3-MONOCHLOROPROPAN-1.2-DIOL <3-MCPD) AFFECTED ERYTHROCYTE MORPHOLOGY AND SEUTROPHIL COU>^T O.V EXPERIMENTAL MICE
Ngo Kien Due, Tran Manh Hung * Y Hoc TPHo Chi Minh * Vol. 14 - Supplement of No 1 -2010:47-51 Backgrotmd: 3-MCPD is fl toxic chemical prodiued by add-h/drolyzed protdn processes. There are bodies of evidence showing toxic effects of 3-MCPD on kidnei/, reproductive organs and cardnogenic property. In tfiis study, we investigated the stibdtronic toxidty of 3-MCPD on erythrocyte morphology and blood cell count.
Method- mice were orally administered 3'MCPD at doses of 1 mg/kg, 10 mg/kg or 20 mg/kg for 6 nionths.
At tlte end cf the study, blood was a)ll&:ted and erythrocyte morpliology was examined. The micronudats test was also perprmed on erythrocyte to investigate the ability of 3-MCPD to induce numerical or structural dtromosomal damage.
Results: 3'MCPD induced changes in erythrocyte morphologi/, induding decrease of size, increase cf pigment and transfi}rming to crenated ceUs. The micronudetis formation in erythrocyte was also increased in 3- MCPD treated groups. These toxic effects of 3-MCPD appeared to be potentiated by tlie presence cf dhanol.
Khoa Dupc - E)?] hoc "t Duac Tp. HCM
Dia chl lien hr: DS. Ngo Kiln Diic DT: 0903 055 .^57 Email: ngokienducg gmail-com
Conclusiom 3-MCPD at doses ofl, 10, or 20 mg/kg, administered in 6 months induced significant changes n erythrocyte morphology and genotoxidty, which may affect tlw fimction oferi/throcytes in drculation.
Keyivcn-ds: 3-MCPD, crenated erythrocyte, micronuclcus, chronic toxicity D A T V A N D E
3-Monochloropropane-l,2-diol (3-MCPD) la mot san phaim phu trong qua trinh san xua't nhieu loai thuc pham. 3-MCPD duoc hinh thanh tir phan ung giira cha't beo va ion Cl' (vi du NaCl) 6 nhiet do cao (Food Standards Agency 2001). Ngoai ra, 3-MCPD ciing duge tao ra trong qua trinh thuy phan protein thuc vat bang a d d hydrochloric (HCI) (Collier 1991). 3-MCPD thuong hi?n dien o ham lugng ra't tha'p (<1 mg/kg) nhung mpt vai loai san pham CO the chiia voi ham lugng cao (len de'n hang tram mg/kg).
Cac nghien cuu tren thii vat cho tha'y, 3- MCPD la chat eo tiem nang gay ung thu tren nhieu eo quan khac nhau 6 chuot cong chiing F344 (Sunahara 1993, Lynch 1998). Nghien cuu in vitro cung cho tha'y 3-MCPD cho ke't qua ducmg tinh tren thir nghi?m gay dot bie'n gen (Silhankova 1982, Zeiger 1988; Lynch 1998).
Mae dil da co ra't nhieu nghien ciru ve dgc tinh cua 3-MCPD, tuy nhien cac dii lieu ve dgc tinh eua 3-MCPD tren mau van chua dugc day du.
Trong nghien cuu nay, chiing foi d?t muc tieu khao sat dgc tinh ciia 3-MCPD tren chuot nhat sau khi cho uo'ng 3-MCPD trong 6 thang lien tyc voi cac chi tieu danh gia la hinh thai hong cau va svr tgo vi nhan.
PHU'ONG PHAP NGHIEN CUtJ Thii vat thu nghidm
Thii vat thu nghiem la chugt nhat trang, chiing Swiss albino, giong due, co trgng lugng tir 18-20 g, dugc phan bo ngau nhien thanh nhieu 16 khac nhau, moi 16 tu 6-20 con. Chupt dirge nuoi trong moi truong tien hanh thuc nghiem tit 3-5 ngay de thich nghi voi moi trirong. Hang tuan, chupt dugc theo doi the trpng va lupng nuoc tieu thu de dieu chinh lugng 3-MCPD thieh hgp. Ke't thuc giai doan thi nghiem, chupt dugc lay mau de phan tich.
Chupt thi nghiem dupc chia thanh cac nhom sau
- N h o m chung: uo'ng nuoc sinh hoat trong 6 thang, n = 18
- N h o m 1: uo'ng 3-MCPD 1 mg/kg/ngay pha trong nuoc u6ng sinh hoat trong 6 thang, n=18
- N h o m 2: uong 3-MCPD 10 mg/kg/ngay pha trong nuoc u6hg sinh hoat trong 6 thang, n=n
-Nhom 3: uong 3-MCPD 20 mg/kg/ngay pha trong nuoe uohg sinh hoat trong 6 thang, n=14
- N h o m 4: uong ethanol 1% pha trong nuoc uo'ng sinh hoat trong 6 thang, n = 6 - N h o m 5: uong ethanol 5% pha trong nuoc
uo'ng sinh hoat trong 6 thang, n = 7 - N h o m 6: uong ethanol 1% - 3-MCPD 1
mg/kg/ngay trong 6 thang, n = 6
-Nhom 7: uong ethanol 5% + 3-MCPD 1 mg/kg/ngay trong 6 thang, n = 6
- N h o m 8: uong ethanol 1% + 3-MCPD 10 mg/kg/ngay trong 6 thang, n = 8
- N h o m 9: uong ethanol 5% + 3-MCPD 10 mg/kg/ngay trong 6 thang, n = 7
Phan tich hinh thai hong cau
De phan tich hinh thai hong cau, mau dugc phe't tren lam kinh, co dinh bang methanol, de kho, roi sau do nhupm mau bang thuo'c thu Giemsa.
Quan sat su hinh thanh vi nhan Thu nghiem nay duoc thie't ke' nham danh gia cac hoa cha't co kha nang gay ton thuong nhiem sac the "Vi nhan" la mpt nhan nho hinh thanh va xua't hien ben eanh nhan binh thuong.
Trong qua trinh phan bao, nhiem sac the se nhan d6i va sau do phan chia vao 2 te' bao eon. Neii tie'n trinh nay bi gian doan, hoac nhiem sac the bi pha vo boi hoa chat hay biic xa thi su phan bo
nhilm sic the vao 2 te'bao con bi anh huong \'a mgt vi nhan se hinh thanh do khong dugc b'ch hpp vao nhan chung. Hien hrgng nay co the dugc quan sat duoi kinh hien vi sau khi nhupm mau nhan bSng thuoc thu duch hgp (Witt 2008).
Phan tich thong ke
Su dung phep phan tich mau Bartiett ve tinh phan bo eiia dii lieu. Neu dii lieu phan bo bmh thuong, sii dung phep kiem ANOVA, ne'u du lieu phan bo bat thuong, sir dung phep kiem Kruskal-WalUs. So sanh su khac biet giiia 2 nhom dugc xem la co y nghia tho'ng ke khi P < 0,05.
KET QU A VA BAN LUAN
Lupng nuoc tieu th»i 6 nhom chung va nhom uong 3-MCPD
Trong suo't qua trinh thii nghiem, chiing t6i theo doi the trpng va lugng nuoc uong trung binh eua chupt (trong nhung nhom nho tii 4-6 chupt) theo timg ngay va tung tuan de CO the dieu chinh Ivrgng 3-MCPD diing theo lieu lirgng thie't ke' nghien euu. Ke't qua duge trlnh bay 6 hinh 1,
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Hinh 1. Liegng nmc tieu thu vd tlie trgng dta chugt a cdc lo thi nghiem
Hinh thai hong cau a nhom chung va nhom uong 3-MCPD
Sau 6 thang su dung 3-MCPD, hinh thai hong eau 6 cac 16 SLI dung 3-MCPD co nhung thay doi dang ke. O nhom chiing, hinh thai hong cau hoan toan binh thuong voi dang hinh dia, sac to' binh thuang. Trong khi o nhom su dung 3-MCPD 1 mg/kg, hong cau mat di dang hinh dia va gia tang sae to'. O nhom Sli dung 3-MCPD 10 mg/kg va 20 mg/kg, ehiing toi nhan thay hong cau chuyen
b-ani; dang hinh cau gai va bat mau ra'l dam (hinh 2).
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Hinh 2. Hmh thai hdng can d nhom chirng va nhom Houg 3-MCPD (x 1000!
S u h i n h t h a n h vi n h a n 6 cac n h o m uong 3-MCFD
O hinh 2, chiing toi quan sat tha'y co su hinh thanh vi nhan 6 cac nhom uo'ng 3-MCPD (mui ten) trong khi o nhom chitng, hau nhu ra't hiem gap hong cau co \ i nhan So lugng \'i nhan tang cao theo nong do 3-MCPD sir dung, giua lieu 1 mg/kg va lieu 10 mg/kg. Sir hinh thanh vi nhan chung to 3-MCPD gav ton thuong nhiem sac the tren cac hong cau luoi Ke't qua dupc trinh bay trong bang 1.
Bdng 1. So'luong vi nhan hinh thanh tren 1 thi tnrdng quan sdt (x 1000 ldn)
Nhom
So hong cau CO VI nhan/!6ng so
hong cau quan sat trong 1 thi t r y o n g
°a hong cau c h u a VI nhan
^*p < 0,01
0/176 1/254 0/127 0/163 0/181 1/205 0.18 ± l!.()l
* " p < 0
3-MCPD 1 mg/kg (n = 6) 3/114 15/177 7/124 13/111 5/119 20/244 6,81 ± 1,35"
0 0 1 SO V 3-MCPD 10 mg/kg
(n = 6) 126/136 213/216 131/233 86/113 100/207 110/140 73,30 ± 2,26™
31 n h o m c 3-MCPD 20 mg/kg (n = 6) 176/230 134/192 36/105 49/102 85/110 74/102 63,321 7,01™
hiing
So l u o n g h o n g cau va cac chi so lien quan 6 n h o m chumg va cac nhom s u diing 3- MCPD
Mac dil 3-MCPD gay nhirng thay doi dang ke tren hirih thai hong cau va su tao vi nhan, tuy nhien khi xae dinh tong lupng hong eau (RBC) giiia nhom chung va nhom sii dung 3- MCPD, chiing toi nhan tha'y 3-MCPD khong anh huong deh tong lugng hong eau va ham lupng hemoglobin (Hb). O nhom sii dung 3- MCPD 1 mg/kg, CO su giam nhe ham luong hematocrit (Het) va the tich trung binh ciia hong cau (MCV) nhung dieu nay kh6ng xay ra tren cac nhom sir dung 3-MCPD 10 mg/kg hay 20 mg/kg (hinh 3). Tren chi so MCHC (nong dp huye't sac to'trong hong eau), eac nhom su dung 3-MCPD deu tang so voi nhom chung (hinh 4).
Htm luvng Hemoglaliln
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aam men
Hiidi 3. Sd'lttgng Itong cdu vd cdc dii solien quan cua cdc Id thi nghiem
Hinh thai hong cau va s u tao vi nhan 6 chupt dupc cho uong dong thoi 3-MCFD va ethanol
Do 3-MCPD duge khii dgc bang phan iing lien hgp voi glutathion de tao thanh S-(2,3- dihydroxypropyl)cystein va N-acetyl-S-(2,3- dihydroxypropyl)cystein (Jones 1975). Ngoai ra, khi sii dung ethanol keo dai co the ga}- suy
giam ham lugng glutathion, vi the ehiing t6i nghien euu trong dieu kien co sir hien dien cua 1 ye'u t6'nguy eo (ethanol), lieu dpc tinh eua 3- MCPD CO gia tang hay khong.
Ve mat hinh thai, viee sii dung do'ng thoi 3- MCPD (10 mg/kg) va ethanol (1% hay 5%) cung gay hien tupng hong eau gai ro ret, trong khi nhom chiing va nhom su dung ethanol co hinh thai hong cau binh thuong. Ve su tao vi nhan, sir dung ke't hgp 3-MCPD 10 mg/kg + ethanol 5% lam tang ty le hong cau co vi nhan len de'n khoang 90% (bang 2).
Bdng 2. Hinh thai hong cau va ty le tao vi nltdn a cdc nhdm thi nghiem
Nhom Chi>ng(n=15) Emanol1%(n = 6) Ethanol 5% {n = 7) Ethanol 1%+3-MCPD
1 mg/kg (n= 6) Ethanol 5%•^3-MCPD
1 mg/kg (n= 7) Ethanol 1% + 3- MCPD10 mg/kg {n=8}
Ethanol 5% + 3- MCPD 10 mg/kg (n=
7)
tfinhthai Binh thuang Binh thiro'ng Binh thu'd'ng Hong cau tang sic H6ng cau tang sic H6ng d u gai Hong c^u gai
°o hong clu CO vi nhan 0,18 + 0.01 0,67 ± 0,31 3.39 + 2,94 8,29 ± 7,9 7,29 ±3,89 41,72±
19,87"
86,99 ± 4,45""
**p <0,01; "•**p <0,001 so voi nhom chiing KETLUAN
Cho de'n nay, da co mpt so' nghien ciiu ve dpc tinh cua 3-MCPD tren mau da c6ng bo', va nhiing nghien cuu nay sii dung 3-MCPD 6 lieu tha'p nhat la 30 mg/kg trong 4-14 tuan. Cac ke't qua cho tha'y 3-MCPD gay thie'u mau, suy tiiy va giam bach eau (Kirton 1970, Marehesini 1989, Cho 2008). Trong nghien a i u ciia chiing t6i, 3-MCPD khong gay thie'u mau nhung gay giam nhe the h'ch hong cau va gia tang nong do huye't sac to'. Dieu nay cho tha'y su thay doi hinh thai ho'ng cau quan sat dugc khong lien quan de'n tinh trang tan huye't va su tao hong cau.
Hong cau gai quan sat dupc trong nghien Cliu ciia chiing t6i, mac du chua xae dinh dupc y nghia benh hpc that sir eiia hien tuong nay.
nhung dieu nay co the lam thay doi ehiic nang cua hong cau va co the gay nhiing bie'n chiing ba't lgi ve huye't dpng.
Ngoai ra, 3-MCPD 6 lieu 1 mg/kg da gay hien tupng tao vi nhan ro ret so voi nhom chiing. Binh thucmg, h5ng cau truong thanh va luu thong trong mau ngoai vi la nhiing te' bao kh6ng nhan. Su tao vi nhan tren hong cau ciia 3-MCPD cho tha'y hoa chat nay tac dpng len rihiem sac the eiia nhiing te' bao tien hong cau va CO the gom ca hong cau luoi.
TAI LIEU THAM KHAO
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Chem Toxicol 46-1666-1673
2 CoUier PD, Cromiue DDO, k Davies AP (1991) Mechanisms of fonnation of chloropropanols present in protein hydrolysates J Am. Oil Chem Soc., 68,785-790 3 Food Standards Agency- (2001)- Survey of 3-
monochloropropane-I,2-diol (3-MCPD) In soy sauce and related products N o 14AH, Food Standards Agency 4 Jones AR (1975) The nwtabolism of 3-chloro-, 3-bromo-
and 3-iodopropan-l,2-diol in rats a n d mice. Xenobiotica, 5, 155-165
5 Jones AR, Milton DH k Murcott C (1978) The oxidahve metabolism of alpha-chlorohydnn in the male rat and the formation of spermatocele. Xenobiotica, 8, 573-582 6 Lynch BS, Br^'ant DB, Hook GJ, N e s t m a n r ER k Mimro IC
(1998) Carcinogenicity of morochloro-1.2-propanediol (alpha-chlorohydnn, 3-MCPD) Inl J, Toxicol, 17,47-76 7 Marehesini M & Stalder R (1983) Toxiaty of 3-ch!oro-l,2-
propanediol m a 4 weeks gavage study on rats. Part 1 Unpubhshed report N'o LA 70/1082 from the Sodete d'Assistance Technique Pour Produits Nestle SA, Switzerland
8 Marehesini M, Stalder R & Perrin 1 (1989) Subchronic toxidty of 3-chloro-I.2-propanediol, 90 days admmistration m drinking water of Fischer F344 rats.
Unpublished report No 1264 from Nestec Ltd Research Centre, Nestle, Switzerland
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10 Sunahara G, Perrin I k Marchesmi M (1993) Cardnogemdty study on 3-monochloropropane-l,2-diol (3-MCPD) admmistered in d n n k m g water to Fischer 344 rats. Unpublished report N o RE-SR93003 submitted to Vmo by Nestec L t d Research & Development, Switzerland
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