Effects of N-acetyl-L-cysteine on Teratogenicity of Cadmium in Mice

ABSTRACT Embryotoxicity and teratogenicity of cadmium (Cd) and modulation of its effects by N-acetyl-L-cysteine (NAC) were evaluated in mice. Pregnant ICR mice were intraperitoneally injected with 3.5 mg/kg of CdCl₂ on day 10 or 11 of gestation (vaginal plug = day 0). Pregnant mice were pretreated with 160 mg/kg of NAC intravenously 2 hours before dosing with CdCl₂. Pregnant mice were killed on day 17 of gestation. Fetuses were examined for external malformations, especially limb malformations, cleft palate and abnormal palatal rugae. There was little difference in body weight gain of dams during the gestation period in the groups treated with NAC plus Cd as compared with the groups treated with Cd alone. Pretreatment with 160 mg/kg of NAC decreased the fetal mortality, incidence of cleft palate and abnormal palatal rugae induced by Cd on day 11. On day 10, pretreatment with NAC decreased the incidence of Cd induced abnormal palatal rugae. These results clearly indicate that NAC exerts protective effects against embryotoxicity and teratogenicity of Cd.     

杂多化合物九钨三钛硅酸盐致畸毒性研究

目的 :观察九钨三钛硅酸盐 ( α- K8H2 〔Si W9Ti3 O4 0 〕· 1 5H2 O,简称 α- Ti3 )孕期给药对大鼠胎仔的毒性影响。方法 :大鼠孕期以 57.0 9,1 71 .2 8和 51 3.83mg/ kg剂量经口给予α- Ti3 ,并以Vit A为阳性对照及蒸馏水为阴性对照 ,产前剖腹检测胎鼠生长发育指标、外观内脏骨骼形态等。结果 :三个剂量组均未见胎鼠外观、内脏及骨骼畸形。低剂量组与高剂量组的活胎率升高 ,吸收胎数下降 ,与阴性对照组比较有显著性差异 (均为 P<0 .0 5)。低、中、高剂量组的胎鼠体重和身长高于阴性对照组 (分别为 P<0 .0 5,P<0 .0 1 ,P<0 .0 1 ) ,且增长程度与剂量呈正相关 ( r=0 .840 8,0 .72 2 8,P<0 .0 0 5,0 .0 5)。低、中、高三个剂量组的上枕骨骨化程度及胸骨块数高于阴性对照组和阳性对照组 ( P<0 .0 5,0 .0 1 ,0 .0 1 ,0 .0 5和 P<0 .0 5,0 .0 1 ,0 .0 1 )。各剂量组的母鼠肝脏重量及肝体比值不同程度下降。其中 ,中、高剂量组及阳性对照组的肝重及肝体比值与阴性对照组比较差异显著 (肝重比较分别为 P<0 .0 5,P<0 .0 5和 P<0 .0 1 ,肝体比值比较均为 P<0 .0 5)。结论 :α- Ti3 致畸试验未见胎鼠畸形 ,但可提高胎鼠生长发育指标和骨骼骨化程度。     

The Enantiomers of the Valproic Acid Analogue 2-n-Propyl-4-pentynoic Acid (4-yn-VPA): Asymmetric Synthesis and Highly Stereoselective Teratogenicity in Mice

The teratogenic activities of R( +)- and S( – )-2-n-propyl-4-pentynoic acid (R and S-4-yn-VPA), the enantiomers of the highly teratogenic valproic acid (VPA) analogues (±)-4-yn-VPA, were investigated in mice. The enantiomers were prepared via asymmetric synthesis, each in three steps employing the chiral auxiliaries (4R,5S)-4-methyl-5-phenyl-2-oxazolidinone and S-4-benzyl-2-oxazolidinone. The determination of the absolute configurations and the optical purities is described. R( + )-4-yn-VPA contained 7%, and S( – )-4-yn-VPA 8%, of the respective antipodes. The aqueous solutions of the sodium salts of R- and S-4-yn-VPA were administered as single i.p. injections during early organogenesis in the mouse (day 8 of gestation) using the induction of exencephaly as the teratological end point. Dose/exencephaly curves indicated that S-4-yn-VPA is 7.5 times more teratogenic than its antipode, 1.9 times more teratogenic than (±)-4-yn-VPA and 3.9 times more teratogenic than the parent drug VPA. In contrast, the neurotoxicity (maternal toxicity) of the 4-yn-VPA enantiomers was found to be independent of the stereo-chemical configuration and lower than achieved after VPA administration. Due to its low neurotoxicity and highly Stereoselective neural tube-inducing activity, S-4-yn-VPA should prove an important tool for the investigation of molecular mechanism of the teratogenic action in this class of compounds; R-4-yn-VPA could act as the negative control in these studies.     

Xa21转基因大米对大鼠致畸作用的实验研究

目的　观察Xa2 1转基因大米对大鼠胚胎生长、发育的影响。方法　将初断乳Wistar大鼠按雌雄分别随机分为 4组 :转基因大米组、非转基因大米组、AIN93G对照组和敌枯双阳性对照组。单笼喂养 ,饲相应鼠料 ,喂满 90天 ,雌雄合笼。观察母鼠和胎鼠的生长发育情况。结果　转基因大米组孕鼠增重、活胎体重、身长、尾长均显著高于阳性对照组 ,而死胎数、吸收胎数、畸形率 (外观、内脏、骨骼 )均显著低于阳性对照组。转基因大米组与非转基因大米组、AIN93G对照组相比 ,所有观察指标均无统计学差异。结论　转Xa2 1基因大米与非转基因大米相比 ,对大鼠受孕率、胚胎生长发育无显著性差异。     

转豇豆胰蛋白酶抑制剂大米的致畸作用研究

目的　用表达杀虫蛋白CpTI(豇豆胰蛋白酶抑制剂 )的大米喂养Wistar大鼠 ,研究其是否具有致畸作用。方法　将断乳的Wistar大鼠随机分为 4组 ,转基因大米组、非转基因大米组、阴性对照组和阳性对照组 ,敌枯双为阳性对照物。转基因大米组含 78 3%转基因大米 ,非转基因大米组含 74 7%与转基因大米同品系的非转基因大米 ,两个对照组的饲料为AIN93G ,三种饲料的宏量及微量营养素的含量相同。大鼠性成熟后 ,进行传统的致畸实验 ,观察母鼠及胎鼠的情况。结果　转基因大米组的孕鼠增重、胎鼠体重、身长和尾长显著高阳性对照组 ,畸形率 (包括外观畸形、骨骼畸形和内脏畸形 )显著低于阳性对照组 (P <0 0 1) ,转基因大米组、非转基因大米组及阴性对照组间的所有指标均无显著性差异 (P >0 0 5 )。结论　此种转CpTI基因的大米对大鼠无母体毒性、胚胎毒性和致畸作用。     

壬基酚对体外培养大鼠胚胎发育的毒性研究

目的 研究4-壬基酚(4-NP)在体外条件下对大鼠胚胎生长发育的影响，探讨其胚胎毒性作用机制。方法 采用植入后全胚胎培养模型，将9.5d Wistar大鼠胚胎与含4-NP的大鼠即刻离心血清共培养48h(4-NP终浓度为0，6.25，12.5，25，50，100mg/L)，观察4-NP对大鼠胚胎生长发育和组织器官形态分化的影响，并用光镜、电镜检测胚胎和卵黄囊的组织结构。结果 4-NP在25mg/L以上可诱发卵黄囊生长和血管分化不良、胚胎生长迟缓及器官形态分化异常，严重者出现神经系统、心脏、腮弓发育异常及小肢芽、体位异常等畸形；100mg/L的4-NP对体外培养胚胎主要呈致死效应。光镜可见实验组胚胎多个器官组织出现病理改变，光镜和电镜下可见卵黄囊3层结构受损，以上结果均呈明显的剂量-反应关系。结论 4-NP对体外培养的大鼠胚胎具有-定的胚胎毒性。其胚胎毒性机制可能与4-NP对卵黄囊胎盘的结构和功能的破坏及其直接的细胞毒性有关。     

Assaying embryotoxicity in the test tube: Current limitations of the embryonic stem cell test (EST) challenging its applicability domain

Testing for embryotoxicity in vitro is an attractive alternative to animal experimentation. The embryonic stem cell test (EST) is such a method, and it has been formally validated by the European Centre for the Validation of Alternative Methods. A number of recent studies have underscored the potential of this method. However, the EST performed well below the 78% accuracy expected from the validation study using a new set of chemicals and pharmaceutical compounds, and also of toxicity criteria, tested to enlarge the database of the validated EST as part of the Work Package III of the ReProTect Project funded within the 6th Framework Programme of the European Union. To assess the performance and applicability domain of the EST we present a detailed review of the substances and their effects in the EST being nitrofen, ochratoxin A, D-penicillamine, methylazoxymethanol, lovastatin, papaverine, warfarin, β-aminopropionitrile, dinoseb, furosemide, doxylamine, pravastatin, and metoclopramide. By delineation of the molecular mechanisms of the substances we identify six categories of reasons for misclassifications. Some of these limitations might also affect other in vitro methods assessing embryotoxicity. Substances that fall into these categories need to be included in future validation sets and in validation guidelines for embryotoxicity testing. Most importantly, we suggest conceivable improvements and additions to the EST which will resolve most of the limitations.     

坤泰胶囊对大鼠胚胎和胎仔的发育毒性研究

目的 评价SD孕鼠在致畸敏感期（怀孕第6～15天）ig给予坤泰胶囊溶液对SD大鼠的母体毒性、胚胎-胎仔毒性和致畸性。方法 雌性SD大鼠低、中、高剂量组在怀孕第6～15天内分别ig给予2.5、5、10 g/(kg?d) 坤泰胶囊成品粉溶液，对照组给予双蒸水。试验期内每天观察动物的一般情况和临床毒性表现；记录孕鼠的体质量；在怀孕第20天进行剖腹检查，摘取卵巢和子宫，观察黄体数、着床数、胚胎生存及死亡情况、生存胎仔的外观、性别、体质量、骨骼及脏器发育等指标。结果 孕鼠及胚胎、胎仔发育的上述各观察指标未见明显异常，与对照组比较无显著差异。结论 在本实验条件下，坤泰胶囊成品粉未见明显的母体毒性与胚胎和胎儿发育毒性。     

Food colorant Sunset Yellow (E110) intervenes developmental profile of zebrafish (Danio rerio)

In this study, we tested the teratogenic/embryotoxic potentials of food colorant, Sunset Yellow (E110) using zebrafish embryos as a model. Laboratory‐raised developing embryos of Danio rerio were exposed to graded concentrations (00, 0.1, 1.0, 2.0, 3.0, 4.0, 5.0, 10, 20, 30, 40, 50 and 100 mm ) of E110 from gastrulation stage (~6 hours post‐fertilization [hpf]) up until hatching. The developmental trajectory of each embryo and post‐hatched larva was traced from 24 to 168 hpf. The no observed effect concentration (NOEC), median effective concentration (EC₅₀), median lethal concentration (LC₅₀) and teratogenic index were determined. In the 0.1 mm E110‐exposed embryos, the development proceeded as in controls (NOEC), while, exposure of embryos to 1‐5 mm of E110 led to a decrease in body size, dry body mass of resultant larvae along with appearance of morphological deformities such as, microphthalmia, pericardial edema, yolk sac edema and spinal curvature. Larvae of 10‐50 mm E110‐exposed embryos exhibited increased cellular apoptosis in the cardiac region with significantly declined heartbeats and elevated mortality rates, in addition to the above‐mentioned abnormalities. In the 100 mm exposure group, all embryos succumbed to death within 24 hpf. The NOEC and LC₅₀ recorded were at 0.1 and 42.57 mm respectively. EC₅₀ (96 hpf) recorded for pericardial edema and yolk sac edema was 19.41 and 39.84 mm with teratogenic index quotient 2.1 and 1.06 respectively The study provides direct evidence for the developmental toxicity/teratogenic potential of E110.