围产期双酚A暴露对子代大鼠的肾脏毒性研究

发布时间：2018-04-17 01:09

  本文选题：双酚A + 围产期　； 参考：《华中科技大学》2012年硕士论文

【摘要】：目的：双酚A（Bisphenol A, BPA）是一种典型的环境内分泌干扰物，主要应用于食品包装行业和牙科，如用于生产聚碳酸酯和无数塑料制品，包括光盘，食品内衬，热（传真）纸，安全帽，粘合剂，粉末涂料等。它的广泛使用，对环境造成了严重的污染，对人类的健康也造成了极大的威胁。已有研究显示，在青春发育期，对SD雄性幼鼠进行BPA的亚急性暴露，可造成大鼠的肾脏损害，并可导致肾积水。但是围产期ＢＰＡ暴露是否可导致子代大鼠肾脏的损害还不清楚。本研究采用了围产期ＢＰＡ暴露的大鼠动物试验模型，研究ＢＰＡ对子代大鼠肾脏损害的影响。方法：在整个妊娠期和泌乳期，怀孕的Wistar母鼠采用随机的方法分为三组：对照组（玉米油）、低剂量组（50μg/kg/day BPA）、高剂量组（250μg/kg/day BPA）。三组大鼠分别接受各组别剂量的灌胃染毒，直至断乳后，子代接受常规饮食至15周。监测子代的体重以及其他生理指标的变化。于第15周时收集尿液、断头处死各组仔鼠，收集肾脏等主要脏器。用生化分析仪、ＨＥ染色、Masson染色、实时定量PCR、免疫印迹、免疫组织分析等技术分别检测尿肌酐（creatinine，CR）、尿素氮(ureanitrogen, BUN)、尿蛋白（Urine protein, UPE）等生化指标、肾脏的病理学改变、肾积水纤维化相关分子TGF-β1、Smad2、VEGF、EGF、ET-1、CD40、CD3、Ⅳ型胶原蛋白等的mRNA的表达水平及蛋白水平的改变。 结果：哺乳期间，BPA暴露组子代体重与对照组相比，变化没有差异。断乳后，至第15周处死时，子代雌性大鼠的体重和肾重及肾脏系数与对照相比，差别未有统计学意义，而子代各剂量组雄鼠与对照组相比，体重增加，有统计学差异。第15周龄各剂量组子代雄鼠的肾重及肾脏系数与对照组相比，有减小趋势，但未有统计学差异。各剂量组雌性大鼠的UPE、BUN、CR差异均无统计学意义，而雄性大鼠的UPE和CR与对照组相比显著升高。而BUN与对照组比较无统计学差异。各组雄性大鼠的肾脏切片呈现一定的病理变化，包括肾实质变薄，肾间质增宽，肾小管扩张、肾小囊扩张，部分肾小球结构破坏，，失去正常形态，伴细胞浸润。各剂量组雄性子代大鼠肾脏纤维化相关分子TGF-β1及其下游调控分子Smad2的ｍRNA表达增强，VEGF的ｍRNA表达显著减弱。同时，免疫印迹结果也显示，TGF-β1、Smad2、VEGF的蛋白表达量也有相应的变化趋势。各剂量组EGF的ｍRNA表达量下降，但未有统计学差异。低剂量组ET-1ｍRNA表达量显著升高，有统计学意义。免疫组化的结果显示，剂量组肾间质CD3+T淋巴细胞较对照组增多，低剂量组肾小管上皮细胞CD40的表达也显著增强。各剂量组IV型胶原在肾小管上皮细胞胞浆和基底膜呈阳性表达。结论：围产期（50μg/kg/day、250μg/kg/day）BPA暴露可影响成年子代雄性大鼠的肾脏功能，其肾脏出现积水和纤维化样改变。围产期BPA暴露对子代雄性大鼠肾脏的影响存在剂量效应关系。
[Abstract]:Objective: bisphenol A bisphenol A (BPAA) is a typical environmental endocrine disruptor, mainly used in food packaging industry and dentistry, such as the production of polycarbonate and countless plastic products, including optical discs, food lining, hot (fax) paper, safety helmet, etc.Adhesives, powder coatings, etc.Its wide use has caused serious pollution to the environment and great threat to human health.Studies have shown that subacute exposure to BPA in young SD rats during puberty can cause kidney damage and hydronephrosis in rats.But it is unclear whether perinatal BPA exposure can lead to kidney damage in offspring.In this study, the effects of BPA on renal damage of offspring rats were studied by using the animal model of perinatal BPA exposure.Methods: during pregnancy and lactation, pregnant Wistar rats were randomly divided into three groups: control group (corn oil), low dose group (50 渭 g/kg/day) and high dose group (250 渭 g/kg/day).Three groups of rats were given intragastric administration of each group until weaning, and their offspring were fed a regular diet for 15 weeks.The body weight and other physiological indexes of the offspring were monitored.Urine was collected at week 15, rats were killed and kidney organs were collected.The biochemical indexes, such as urinary creatinine creatinine, urea nitrogen, bun, urinal protein, UPEN, and the pathological changes of kidney were detected by biochemical analyzer, real-time quantitative PCR, immunoblotting, immunohistochemistry and other techniques, such as ureanitrogen, ureanitrogen, urinal protein, UPEN, and so on.Changes of mRNA expression level and protein level of TGF- 尾 _ 1 Smad2 + VEGFT-1 CD40 CD40 CD3, 鈪

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