J. Fd Hyg. Safety 21(4), 197 203 (2006) q ƒ f e x ½ 1 Áy 1 Á 1 Á z 1 Á 1 Á x 1 Á x 1 Á 1 Á 1,3 Á 2 Á 1,3 Á 1 w wœ w œ w w wp w w œm Assessment of Pubertal Development to Parabens-induced Estrogenic Effect in Male Mice Sun-Jung Kim 1, Jae-Woong Hwang 1, Jung-Ran Park 1, Yeong-Geon Lee 1, Ji-Hye Chung 1, Yun-Hyeok Jeong 1, Soo-Jin Lee 1, Ji-Won Jung 1,3, Ji-Youn Jung 2, Yong-Soon Lee 1,3, and Kyung-Sun Kang 1 %FQBSUNFOUPG7FUFSJOBSZ1VCMJD)FBMUI$PMMFHFPG7FUFSJOBSZ.FEJDJOF4FPVM/BUJPOBM6OJWFSTJUZ4FPVM,PSFB %FQBSUNFOUPG$PNQBOJPOBOE-BCPSBUPSZ"OJNBM4DJFODF,POHKV/BUJPOBM6OJWFSTJUZ$IVOH/BN,PSFB,3'QSJPSJUZ3FTFBSDI*OTUJUVUFGPS;PPOPUJD%JTFBTF 3FDFJWFE/PWFNCFS"DDFQUFE%FDFNCFS ABSTRACT Parabens are most wildly used in food, cosmetics and pharmaceutic products as preservatives caused of safety and cheap. we had examined that paraben had estrogenic activity through the in vivo and in vitro experiments in last year. We demonstrated that most of parabens(ethyl, butyl, propyl, isobutyl, isopropyl) increased significantly uterus weight as well as induced proliferation of MCF-7 cell and binding of estrogen receptor as endocrine disrupter compounds. In this study, we evaluated that whether parabens have effect on male reproductive system or not. the male rats were administrated parabens by oral injection then examined separation of preputial day for PND23~PND52. As the results, most parabens delayed pubertal development compare to control group. The separation of preputial day of Butyl and Propyl parabens at high concentration were PND 44 days and PND 45days compared to control group as PND 40 days. Even though, parabens as endocrine disrupter wildly spread in food, cosmetics and pharmaceutic products, we didn't have the safe guideline. In abroad, they are re-evaluating safety assessment for parabens. In conclusion, parabens delayed pubertal development in juvenile. parabens are consider as endocrine disrupter chemicals. Key words: parabens, estrogenic activity, endocrine disrupter, preputial day y y y ü š w, y yw ü š, y w w. ü w, x ¾ 70 j w yw š. 1990 ü w w š ƒ» w, y ww ù g ù f y ³x w y w w š. Author to whom correspondence should be addressed. ü q w ƒ, w, y t, t, t. p, v v, v v, p, p. q y w p ƒ š, p w ü w 1,2). Dabre et al. y q ƒ x ƒ š w, w q n w F1 male š. 3) q q ƒ f ª 4) w in vitro x s 197
198 4VO+VOH,JNFUBM MCF-7 s ƒ jš, p ww š w 5). xw û w w e w» w, w q ( p, v v, v v, p, p) SD f, PND 23 l q x ù m PND 52 ¾ n w, n»,sv» wš, n z w šy, û, û» y y d ü w. x x p q, v v q, v v q, p q, p q (Corn Oil), v k (Flutamide) 1) w. x,,, x Sprague Dawley(( ) p g :» û 1 143-1 )) w f f Sprague Dawley rat ƒƒ 20, w z yw w z 18 f d w z s³ ±5% ü w w wš 2 z( z 20 ) d w ±5% ü w ( n ), (Flutamide) pq n, v vq n, v vq n, pq n, p q n ƒƒ 10 x w. n n n k x 4ml/kg z 23 (PND 23) l 52 (PND 52) ¾ 1 1z 30 n w.,, d, d n» 1 1z k y,, w. w 1 1z ù k vš, k ƒ w(autolysis)» w ether g ww. x s w e wš, ƒ w þ z 24 ü w. n 2 ( z 21 ), n ( z23 ), n» d w. n» d w. sv d tƒ sv d w, w PND 23 l n ¾ w, sv 2-4) d» w. y (TSH. T4,T3, Testosterone) d w n z (abdominal aorta) l xw x l TSH, T4, T3 Testosterone d w.,» d n z ether g w z, l x e g ü» w w z šy(testes), šy (Epidiymides), (Prostate), û(seminal vesicle with coagulating gland), LABC(Levator ani plus bulbcavernosus muscle complex), (Thyroid), w (Pituitary gland), (Liver), (Paired Kidney) w» d wš w» w. w x (Testes), šy(epidiymides), (Prostate), û(seminal vesicle with coagulating gland), LABC(Levator ani plus bulbcavernosus muscle complex), (Thyroid), w (Pituitary gland), (Liver), (Paired Kidney) w 10 % s (10% buffered neutral formalin) š wš w Ì (3 mm) w, q v s w 4 µm r w. z Hematoxylin & Eosin (H&E stain) w Ÿw x (Olympus BX50, Olympus Optical Co., Japan) w w ww. m x d x w m w w one-way ANOVA w p=0.05 wš, Dunnett's t-test ww x m w w (p<0.05).
"TTFTTNFOUPG1VCFSUBM%FWFMPQNFOUUP1BSBCFOTJOEVDFE&TUSPHFOJD&GGFDUJO.BMF.JDF 199,, d, d» x,, (v k ) p w. w n» w x e w. wr d w v k w q e w (Fig. 1). û w ù m w ( ). sv d q ƒ f e w xw, sv 40±1.83, ƒ sv r ƒ v v q, š (2000 mg/kg/day) 44±1.94. p q ƒ š 2000 mg/kg/day sv 44±2.73. w p Fig 1. Body weight Changes (A) Control, (B) Ethyl Paraben, (C) Butyl Paraben, (D) Isobutyl Paraben, (E) Propyl Paraben, (F) Isopropyl Paraben.
200 4VO+VOH,JNFUBM q (2000 mg/kg) 43±1.25 s v. v k (100 mg/kg/day) ú(pnd 53) sv, x v k sv 53-56. y (TSH. T4,T3, Testosterone)d T3(ng/dL) (66Û4.73 ng/dl) w v k (50Û6.75 ng/dl), q w w ùkü. ƒ v v q 2000 mg/kg/day (53.4Û8.69 ng/dl) v v q 2000 mg/kg/day (48.5Û8.02 ng/dl) ùkü. T4(ug/dL) (3.9Û0.61 ug/ dl) w v k sww q w w, e q ƒ w w. p pq š 2.7±0.77 ug/dl. w v v q š (2000 mg/kg.day) e 2.42 ±0.59 ug/dl T4 y eƒ û, j v v š 1.87Û0.70 ug/dl ƒ. TSH(ul/dL) y v v q wš v k sww q w m w. Testosterone, v k w q w û w ù, m w (Fig. 2).» d w šy š Fig 2. Hormone Level (A) T3 (ng/dl) Hormones (*P<0.05), (B) T4 (ug/dl) Hormones(*P<0.05), (C) TSH (ul/ml) Hormones (*P<0.05), (D) Testosterone Hormones (ng/ml) (*P<0.05).
Assessment of Pubertal Development to Parabens-induced Estrogenic Effect in Male Mice 환, 전립선 정낭 등에서 위축이 보였으며, 파라벤류처치군 에 서도 고농도 군에서 고환 및 부고환, 전립선 정낭 등에서 위 축을 보였다. 부검 후 장기 무게 결과, 양성 대조군인 플루 타마이드군에서 고환, 부고환 전립선, LABC, 정낭 무게가 유의적으로 감소했으며, 전반적으로 모든 파라벤류에서 정 소, 부고환, 전립선, 정낭, LABC, 뇌하수체, 간 등의 절대 장기 중량이 유의적으로 감소하였다. 체중 대비 장기 중량을 보면 양성대조군인 플루타마이드 처치군 에서 정낭의 상대 장기 중량이 유의적으로 감소하였으며, 파라벤류 처치군 에 서는 부틸(2000 mg/kg/day)와 에틸(1000 mg/kg/day)에서 전립선의 상대 장기중양이 유의적으로 감소하였다. 다른 파 라벤류 처치군 에서는 그 유의성을 찾을 수 없었다. 조직 병리학적 소견 조직 병리학적 결과, 대조군에 비해 양성 대조군(Fig. 3(B), Fig. 4(B))은 고환과 부고환에서 유의적인 위축을 보였 으며, 부고환 내강에 정자가 결여 된 것도 확인되었다. 대부 분 파라벤류에서는 상대 장기 중량에 대한 통계학적 유의성 은 없었으나, 조직 병리학적으로 고환과 부고환의 위축이 관 찰되었다. 특히 부틸파라벤(Fig. 3(C), Fig. 4(C))과 프로필 파라벤(Fig. 3(D), Fig. 4(D)) 고농도군(2000 mg/kg/day)에 서 고환과 부고환의 위축이 관찰되었으며, 프로필 파라벤의 경우 강 내 정자가 결어된 것도 관찰되었다. 모든 파라벤류 가 농도 의존적으로 고환과 부고환의 위축됨이 관찰되었으 나 고환보다는 부고환이 더 유의적으로 위축됨이 관찰되었 다. 또한 전립선의 경우 대조군(Fig. 5(A))에 비해 부틸 파 라벤과(Fig. 5(C)) 프로필 파라벤(Fig. 5(D)) (2000 mg/kg/ Fig 3. Testes (A) Control, (B) Positive Control (100 mg/kg/ day), (C) Butyl Paraben (2000 mg/kg/day), (D) Propyl Paraben (2000 mg/kg/day). 201 day) 농도 군에서 Hyperplasia가 관찰 되었다. 또한 상대 장 기에서 전립선이 유의적으로 감소하였던 에틸 파라벤(Fig. Fig 4. Epididymides (A) Control, (B) Positive Control (100 mg/kg/day), (C) Butyl Paraben (2000 mg/kg/day), (D) Propyl Paraben (2000 mg/kg/day). Fig 5. Prostate (A) Control, (B) Positive Control (100 mg/kg/ day), (C) Butyl Paraben (2000 mg/kg/day), (D) Propyl Paraben (2000mg/kg/day), (E) Etyl Paraben (1000 mg/kg/day). Journal of Food Hygiene and Safety, Vol. 21, No. 4
202 4VO+VOH,JNFUBM 5(E)) 1000mg/kg/day p v v q j Hyperplasiaƒ w. q ƒ e w» w x ww,, q sv ƒ, p v v q p q j g. w šy š y w x w, hyperplasia ƒ» ww y w. š q ƒ f k w. p v v q pq š (2000 mg/kg/day) 4 sv g. (s³ 44 ) w y e T4 y p q š, T3 v v q š y eƒ w û. ù Testosterone, TSH q ƒ ƒw w w ù m w. q e š šy š y, û,» š w p q š pq š x w, q» ù. w q pq ( 1-3, 1-7) v v q ( 1-4, 1-8) š (2000 mg/kg/day) šy šy, v v q ü ƒ. q ƒ šy šy ƒ, šy. w v v q š Hyperplasiaƒ š, w» w p q ( 1-13) 1000 mg/kg/ day p v v q j Hyperplasiaƒ w. û y t ƒ q ƒ t w ƒw q ƒ ü û y šy šy» w. y t t q ƒ w š, ƒ Oishi et al. p w» w. 6,7) 8) p q w ù p v v q f û y l m l w» w ƒ tw. w q 9,10) p y w k x ƒwš. 11), q ƒ ùƒ wš ü»w mw q ƒ û ww xü š,, q ƒ ü y ww w w» k w ƒ. / t t 2005 ü sƒ w w. 2006 w w (KFR-005-E00076). š x 1. Darbre, P.D., Aljarrah, A., Miller, W.R., Coldham, N.G., Sauer, M.J. and Pope, G.S.: Concentrations of parabens in human breast tumours. J Appl Toxicol. 24, 5-13 (2004). 2. Darbre, P.D.: Underarm cosmetics and breast cancer. Eur J Cancer. 13, 153 (2004). 3. Kang, K.S., Che, J.H., Ryu, D.Y., Kim, T.W., Li, G.X., and Lee, Y/S.: Decreased sperm number and motile activity on the F1 offspring maternally exposed to butyl p-hydroxybenzoic acid (butyl paraben).j Vet Med Sci. 64, 227-235 (2002). 4. Jo, E.H., Jung, J.Y., Park, C.B., Park, S.H., Lee, Y.S., and Kang, K.S.: Estrogenic activity of parabens in yterotrophic assay. J of food hygiene and safety. 21, 118-128 (2006). 5. Lee, S.H., Kim, S.J., Park, J.R., Lee, Y.S., and Kang, K.S.: Estrogenic activity of parabens in vitro Estrogen Assay. J of food hygiene and safety. 21, 100-106 (2006). 6. Soni, M.G., Carabin, I.G. and Burdock, G.A.: Safety assessment of esters of p-hydroxybenzoic acid (parabens). Food Chem Toxicol., 43, 985-1015 (2005). 7. Gomez, E., Pillon, A., Fenet, H., Rosain, D., Duchesne, M.J., Nicolas, J.C., Balaguer, P., and Casellas, C.: Estrogenic activity of cosmetic components in reporter cell lines: parabens, UV screens, and musks. 8. Oishi, S., Lack of spermatotoxic effects of methyl and ethyl esters of p-hydroxybenzoic acid in rats. Food Chem Toxicol.
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