1、Associations between phthalates exposure and health outcomes of newborns, pregnant women and precocious puberty girls in Taiwan,李俊璋 教授 國立成功大學工業衛生科暨環境醫學研究所 成功大學環境微量毒物研究中心,2,Whats Phthalates (PAEs)?,PAEs,They are a family of chemical substances that have been in use for about 50 years, primarily to ma
2、ke polyvinyl chloride (PVC) soft and flexible. Phthalates look like clear vegetable oil.,Physico-chemical properties of Phthalates,Applications of phthalates,Personal care products,Application of phthalates,Metabolic pathway of DEHP,-Albro, et al., 1982; ATSDR, 1993, Schmid, et al., 1985c.,DEHP,MEHP
3、,Human,Primary,Secondary,Objectives,Part I: Association between Phthalate Exposure and Thyroid Hormone in Pregnant Women,To determine the concentrations of urinary phthalate monoesters in pregnant women. To investigate the association between phthalate exposure and thyroid hormone during pregnancy.,
4、Objectives,Part II: Association between Prenatal Exposure to Phthalates and the Health of Newborns,To determine the concentrations of amniotic fluid phthalate monoesters in pregnant women. To evaluate the association between prenatal exposure of the fetus to phthalates in amniotic fluid and maternal
5、 urine and the health of newborns.,To follow-up our previously established participants in order to determine the correlation between phthalate exposure and thyroid hormones in pregnant women and newborns.,Objectives,Part III: Phthalates may alter thyroid hormones in utero: A follow-up study,Framewo
6、rk,Newborns health Anogenital distance Gestational ageUrine sample Phthalate monoestersSerum &Cord blood TSH,T3, T4, FT4 Phthalate monoesters,Materials and Methods,Informed consent Amniotic fluid Phthalate monoestersUrine sample Phthalate monoesters Creatinine Serum sample TSH,T3, T4, FT4,Urine samp
7、le Phthalate monoesters CreatinineSerum sample TSH,T3, T4, FT4Questionnaire Age, BMI, gestation Food consumption Cosmetic usage,Follow up-76 subjects,Follow up-65 subjects,Interview- 83 subjects alpha fetal protein (AFP) -free -hCG maternal age 35 yrs undergo amniocentesis,12,Measured Parameters,Pht
8、halate monoesters monomethyl phthalate (MMP) monoethyl phthalate (MEP) monobutyl phthalate (MBP) monobenzyl phthalate (MBzP) mono-2-ethylhexyl phthalate (MEHP) Sample Urine/ Amniotic fluid Serum Analytical method LC-MS/MS (Agilent 1100/API 3000),Samples,Measured Parameters,Physical examination Anoge
9、nital distance (AGD) Birth weight Birth height Gestational age Anogenital index (AGI),AGI-W= AGD/ birth weight. AGI-L= AGD/ birth length.,Thyroid hormones Thyroid stimulating hormone (TSH) Thyroxin (T4) Triiodotyronine (T3) Free thyroxin (FT4) Questionnaires Age, BMI, gestation age, Smoking, Medical
10、 care etc. Food consumption Cosmetic usage Personal care product usage Building characteristics,Table I-1 Demographic characteristics of study participants (n = 76).,Characteristics Mean SD,Age (years) 33.6 3.3 Body mass index (BMI) 20.9 2.5 Duration of gestation (weeks) 27.9 2.3 Pregnancies and bir
11、thsNumber of current pregnancy 1.9 1.0Number of current birth 1.5 0.6 Smoking status (n/%)Active smoker 0/0Passive smoker 14/18.4Non-smoker 62/81.6 Alcohol drinker (n/%) 0/0 New decoration of living/ working place during previous 1 year (n/%) a HomeMoving to new decorated house 7/9.2Just decorated 4
12、/5.3 WorkplaceMoving to new decorated workplace 2/2.6Just decorated 6/7.9 Medical care during previous 3 months (n/%)Blood transfusion 3/3.9Intravenous drip 3/3.9Oxygen mask 1/1.3,a Moving to new decorated house or workplace means moved into a new building. Just decorated means the place you live or
13、 work have decorated recently such as painting or changing the floor.,15,Table I-2 Levels of five urinary phthalates monoester in pregnant Taiwanese women and comparison with other studies (g/g creatinine).,Table I-3 Distribution of thyroid hormones in pregnant women.,aThe analytic sensitivities of
14、T3, T4, free T4, and TSH were 19.5 ng/dL, 0.42 g/dL, 0.023 ng/dL and 0.014 lU/mL, respectively; the coefficient variations of T3, T4, free T4, and TSH were 2.9%, 4.2%, 3.1%, and 3.0%, respectively.,Table I-4 Spearman correlation coefficients a between thyroid hormone, age, BMI, duration of gestation
15、 and urinary phthalate monoester levels (n = 75).,a Spearman correlation coefficients, *: p 0.05.,Figure I-1. Correlation between (A) free thyroxine (FT4), (B) thyroxine (T4) and log MBP (ppb) and MBP-c level in pregnant women at the 2nd trimester.,A,B,Table I-5. Multivariate linear regression betwe
16、en serum FT4 and T4 levels, and their corresponding urinary phthalate monoesters (FT4: R-square= 0.240; T4: R-square= 0.187)a.,a One outlier was excluded because of hypothyroidism (n=75).,Multiple regression: FT4： = -0.110, R2=0.24, p0.05 T4：=-0.112; R2=0.187, p0.05,Table II-1 Physical examination o
17、f male and female newborns (n = 65).,1 The AGD of two girls and boys, respectively, were not available. 2 AGI=ano-genital index; AGI-W= AGD/birth weight; AGI-L=AGD/ birth length. 3 *: p0.05,Table II-2 Concentrations of five phthalate monoesters in amniotic fluid from female and male infants (ng/mL,
18、n=64)1.,1One amniotic fluid sample was failed during analysis. 2ND=not detected, detection limit of MMP, MEP, MBP, MBzP and MEHP were as follow: 1.4, 1.0, 1.4, 1.4, 0.9 ng/mL,Figure II-1 Distribution plot of MBP levels in maternal urine and amniotic fluid (n=59). All the data were log-transformed.,I
19、t is indicated that current levels of exposure to phthalate in pregnant women may only partly represent in utero fetal exposure,Table II-3 Distribution of health status in females categorized by median levels of phthalate monoester in amniotic fluid (-AF) (n = 31).,1Wilcoxon Rank Sum Test, *: p 0.05
20、; 2AGD data were lacking for one female newborn in each group. 3AGD data were lacking for two female newborns in the high MEHP-AF group. 4Median levels of amniotic fluid MBP in low and high group were 67 and 104 ng/mL, respectively; those of MEHP were 9.5 and 38.8 ng/mL, respectively.,Table II-4. Di
21、stribution of health status in boys category by median levels of phthalate monoester in amniotic fluid. (n = 33).,1 One male infant of each group in MBP-AF and MEHP-AF, respectively, were lack of AGD.,Table II-5 Spearman correlation coefficients1 between birth weight, birth length, gestational age,
22、AGD, AGI-W, AGI-L, and phthalate monoester levels in amniotic fluid in female infants (n = 29)2.,1 *:p 0.05; #: p0.06. 2 BW=birth weight, BL=birth length, GA=gestational age, AGD=ano-genital distance, AGI-W=AGD/BW, AGI-L=AGD/ BL,Figure II-2 (A) Linear regressions between MBP in amniotic fluid and AG
23、I-W in females at birth (n=29, R2=0.14, p=0.043); (B) Linear regressions between MBP in amniotic fluid and AGI-L in females at birth (n=29, R2=0.16, p=0.032).,A,B,Negative associations among amniotic fluid MBP and AGI,Figure III-3 Multiple regression of AGI, MBP in amniotic fluid and gestational age
24、 adjustment of gestational age and other phthalate monoesters (amniotic fluid MBP: = 2.73, p = 0.041; gestational age: = 0.899, p = 0.006) .,Table III-1 Concentrations of phthalate monoesters in urine, serum and cord blood at delivery (ng/mL, n=61).,aMBP = monobutyl phthalate; MBzP = monobenzyl phth
25、alate; MEP = monoethyl phthalate; MEHP = mono-2-ethylhexyl phthalate; MMP = monomethyl phthalate. bDetection limit (LOD) of phthalate monoesters were: MBP, 1.4; MBzP, 1.4; MEP, 1.0; MEHP, 0.9; MMP, 1.4 ng/mL, respectively. Half of LOD was calculated as the detected value below the LOD.,Table III-2 C
26、oncentrations of thyroid hormones1 in maternal serum during pregnancy and cord blood at delivery (n=61).,1Reference values for thyroid hormones in Taiwan: TSH: 0.27-4.2; T3: 84.6-202.0; T4: 5.13-14.1; FT4: 0.93-1.7. The analytic sensitivities of T3, T4, free T4, and TSH were 19.5 ng/dL, 0.42 g/dL, 0
27、.023 ng/dL and 0.014 lU/mL, respectively. The coefficient variations of T3, T4, free T4, and TSH were 3.0%, 4.1%, 3.2%, and 2.9%, respectively.,Table III-3 Correlation a coefficients between fetal thyroid hormones and phthalate monoesters in cord blood samples (n=60)a,.,a Spearman correlation: *p0.0
28、1, *p0.05, #p0.1. b-C: Phthalate monoesters in cord blood sample.,Figure III-2 Linear correlation between log MBP levels and TSHT4 in cord blood samples (R2=0.081, p=0.028, n=60).,Table III-4 Multivariate regression between TSH and T4 and their corresponding phthalate monoesters in cord blood (n=60)
29、a,33,Possible Hypotheses,First, we postulated phthalate exposure may affect fetal thyroid function indirectly via decreased thyroid hormones from their mother. Animal studies have shown that in utero exposure to thyroid hormone affects the regulation of the hypothalamus-pituitary-thyroid axis in new
30、borns (Alonso et al., 2007; Piosik et al., 1997). Our previous study showed decreased FT4 and T4 levels with increasing urinary MBP levels in pregnant women during the second trimester (Huang et al., 2007). Second, phthalate monoesters penetrate the human placenta and may have direct effects on feta
31、l thyroid function. Some toxicological studies have shown that certain phthalates, such as DBP and DEHP, are possible thyroxine antagonists that affect the transportation, secretion, and action of thyroid hormones in adult animals (Hinton et al., 1986; Poon et al., 1997; Sugiyama et al., 2005). Alth
32、ough little information is available about phthalate exposure in utero and its effects on fetal thyroid function, some studies (Huang et al., 2007; Meeker et al., 2007) have shown possible negative effects in humans.,性早熟童液中鄰苯二甲酸酯代謝物檢測與家戶灰塵之相關性研究The study on relationship between urinary phthalate mon
33、oesters and house dust exposure in precocious puberty girls,行政院國家科學委員會補助專題研究計畫成果報告,Phthalate in house dust,Table 1. Estimated intakes of phthalates compared with the tolerable daily intake (values in g/kg body weight/day),(Fromme et al., 2007),36,Exposure route of phthalates,INGESTION,SKIN ABSORPTIO
34、N,INHALATION,newlatesttips.info,, prone to be higher exposed Lower body weight Hand to mouth behavior (sucking/ mouthing dust),Schettler, 2006; Sathyanarayana, 2008; Wittassek, 2010,Fig 1 The framework of this study,38,Sampling strategy for dust- 5 samples,Kitchen,bathroom,Parents room,Living room,P
35、layroom,Dust collection equipment,Using aluminum-made collector & cellulose filter Covering with aluminum foil and then sealing by parafilm after collecting enough dust sample,探討居家特性等因子對於兒童室內鄰苯二甲酸酯濃之影響,40,Distributions of phthalate levels in settled dust (n=965) (g/g dust),41,N.D.: non-detection,42,
36、Worldwide comparison of phthalates concentration,ND, no data. a 50th and 95th percentiles. b Mean concentration. c 90th percentile.,Adapted from Kolarik, 2008,灰塵樣本採集高度與鄰苯二甲酸酯類濃度分佈情形,44,DBP level and wooden floor,45,Significantly higher DBP concentration was found at major activity room and above flo
37、or level when using wooden materials for flooring in the room.,Wax using and phthalates concentration,46,Regular waxing was associated with higher BBzP and DEHP levels in dusts collected from the floor.,Parental occupations and DEHP levels,47,父母親從事鄰苯二甲酸酯類相關的職業可能為居家室內DEHP的來源之一,探討環境鄰苯二甲酸酯濃與孩童內在劑（液鄰苯二甲
38、酸酯）之相關性,48,Worldwide comparison of phthalate metabolites concentration,49,Correlation of concentrations between dust-borne phthalates in different place and their corresponding metabolites of childs urine,*p0.05, *p0.01,Correlation of concentrations between dust-borne phthalates and their correspond
39、ing metabolites of childs urine,*p0.05, *p0.01,Dietary pattern and phthalates metabolites,*p0.05,探討鄰苯二甲酸酯與童性早熟發生之相關性及環境控管介入研究,53,Subjects,The present case-control study was carried out from 2006 to 2009. 71 central precocious puberty (CPP) girls were recruited from the policlinic of Pediatric Endocr
40、inology in National Cheng Kung University Hospital and 29 normal similar age-range girls were recruited as control group from elementary school or kindergarten.,Table The levels of seven PAEs metabolite in case and control(g/g-creatinine),Wilcoxon rank sum test; Geometric mean (Range),The analytical
41、 results of urine samples,Results,Table The kisspeptin-54 levels, basal hormones and physical characteristics from part of precocious puberty and normal girls in this study.,Fig The association between Kisspeptin-54 and ln MBP-Cr (g/g-Cr).,R2= 0.288;p= 0.001,Results,Results (Intervention strategies)
42、,介入期程: 一個星期,表 性早熟女童與一般女童介入後濃度下降之族群其尿液未校正creatinine之幾何平均濃度前後差異測值(範圍)(單位：ng/mL)與各種介入方案之關係,表 性早熟女童與一般女童介入後濃度下降之族群其尿液校正creatinine後之幾何平均濃度前後差異測值(範圍) (單位：ug/g-creatinine)與各種介入方案之關係,本土一般住家內DEHP濃度普遍高於歐美國家。家中使用木製地板、有打蠟習慣、父母親在特殊工廠工作等皆會貢獻較高DBP、BBzP及DEHP濃度於灰塵中。 孩童尿液中之MBP、MEHHP及MEOHP濃度明顯偏高，顯示家戶孩童可能受到較多DBP及DEHP之暴
43、露，且與紙盒包裝產品、塑膠瓶/杯容器的使用習慣有關。由於採取介入措施可有效降低尿液中濃度建議政府機關應針對產品製造商進行管制，逐步採用其他危害性較低之塑化劑，或降低此兩類物質在產品中的含量，並避免利用塑膠產品裝盛和保存食物。 性早熟女童尿液中之MEP、 MEHP、MEHHP及MEOHP濃度明顯高於一般女童，顯示性早熟女童可能受到較多DEP及DEHP之暴露。DEP、DEHP多使用於塑膠製品、個人衛生用品及化妝品中，建議兒童應減少塑膠製品及化妝品(如香水、乳霜、指甲油等)的使用 性早熟女童尿液中之MBP與kisspeptin具顯著相關性，顯示DBP暴露可能經由影響kisspeptin而誘發性早熟。
44、,結論與建議,一般民眾如何自我評估暴露程度,一般民眾如何自我評估暴露程度,一般民眾如何自我評估暴露程度,一般民眾如何自我評估暴露程度,要怎麼降低或避免塑化劑的暴露 ?,避免喝塑膠杯裝市售飲料的習慣，儘量自己帶不銹鋼杯或馬克杯。 避免以塑膠袋、塑膠容器盛裝熱食或微波加熱，平常的生活中也減少使用塑膠袋。 養成吃東西前洗手的習慣，避免因手接觸室內灰塵而食入塑化劑。 避免用保鮮膜包覆食物進行微波或蒸煮。 兒童應避免在巧拼地板上吃東西、玩耍，也不要直接睡在巧拼地板上。在巧拼地板上活動後，要保有洗手的習慣，並避免幼童將手放置口中。 室內環境應定期吸塵以維持乾淨，電器不用時應拔除插頭以避免經常在高熱狀態下逸
45、散塑化劑。 多使用含天然成分或不含塑化劑之香水、化妝品及個人衛生用品。 避免長期吃單一種類的高脂食品，例如：可以交替食用豬肉、牛肉、雞肉或以魚類替代，都可以降低塑化劑的可能暴露。 多喝白開水，多攝取新鮮蔬果，少吃高脂食物，並保持心情愉悅,Acknowledgements,We are grateful for the financial support of Taiwan National Science Council and deeply appreciative of the subjects participated in this study. Livestock Research Institute, Council of Agriculture Main-Lian Lee, Ming-Chih Teng Department of Pediatrics Yen-Yin Chou, Shio-Jean Lin Department of Obstetrics and Gynecology Pao-Lin Kao, and other gynecologists Research Center of Environmental Trace Toxic Substances All stuffs,
