Article Summary
赵 圣,李 登,朱依萍,贝晓宇,蒋君涛,韩邦旻.孕期邻苯二甲酸二正丁酯暴露促进大鼠子代肾小管上皮细胞发生Snail1介导的上皮-间充质转化[J].现代生物医学进展英文版,2019,19(12):2207-2213.
孕期邻苯二甲酸二正丁酯暴露促进大鼠子代肾小管上皮细胞发生Snail1介导的上皮-间充质转化
Maternal Exposure to Di-n-butyl phthalate Induces Snail1-mediated Epithelial-mesenchymal Transition in Rat Renal Tubular Cell
Received:January 08, 2019  Revised:January 30, 2019
DOI:10.13241/j.cnki.pmb.2019.12.002
中文关键词: 邻苯二甲酸二正丁酯  上皮-间充质转化  Snail1  肾纤维化
英文关键词: Di-n-butyl phthalate  Epithelial-mesenchymal transition  Snail1  Renal fibrosis
基金项目:国家自然科学基金项目(81771564)
Author NameAffiliationE-mail
ZHAO Sheng Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China medseek@163.com 
LI Deng Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China  
ZHU Yi-ping Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China  
BEI Xiao-yu Institute of clinical transformation, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China  
JIANG Jun-tao Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China  
HAN Bang-min Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China  
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中文摘要:
      摘要 目的:本实验以DBP暴露的大鼠为模型,研究DBP相关纤维化肾脏细胞上皮-间充质转化(EMT)水平改变以及该过程的调节机制。方法:动物模型中实验组妊娠大鼠在妊娠14-18天期间以800 mg/kg/天的剂量胃饲DBP,使用免疫组织化学(IHC)检测EMT相关指标;使用IHC、Western blot和PCR技术检测转录因子Snail1表达;以肾小管上皮细胞NRK52E作为体外模型,使用同样方法检测DBP暴露对中TGF-β1表达影响,检测H2O2对TGF-β1表达影响、TGF-β1信号通路拮抗剂对DBP诱导的Snail1表达的影响。结果:与对照组相比较,孕期DBP暴露导致子代肾脏E-cadherin指标显著升高,IHC染色强度超过对照组的3倍,N-cadherin指标显著下降,IHC染色强度约为对照组20%(P<0.05)。 DBP能够显著促进肾小管上皮细胞Snail1表达,IHC染色强度相比对照组升高约2.5倍(P<0.05),干扰Snail1能够抑制DBP诱导EMT指标的改变;DBP暴露与肾脏TGF-β1高表达相关,TGF-β1信号通路抑制剂能够引起DBP相关的Snail1表达显著降低(约25%);此外,DBP造成的活性氧(ROS)产物累积促进了肾小管上皮细胞TGF-β1的表达(P<0.05)。结论:孕期暴露于DBP会导致肾脏ROS产物累积和TGF-β1高表达,进而促进肾小管上皮细胞发生Snail1介导的EMT。
英文摘要:
      ABSTRACT Objective: In this study, DBP-exposed rats were used as models to study the changes in the level of fibrotic kidney epithelial-mesenchymal transition (EMT) and the regulatory mechanisms of this process. Methods: Pregnant rats in experimental group were fed with DBP at a dose of 800 mg/kg/day during the 14-18 days of gestation. Immunohistochemistry (IHC) was used to detect relevant indicators of EMT; IHC, Western blot and PCR were used to detect expression of transcription factor Snail1; Renal tubular cells NRK52E was used in vitro. With the same method, expression of TGF-β1 in vivo and in vitro, effect of H2O2 on TGF-β1 expression and effect of TGF-β1 receptor inhibitor on DBP Inducted Snail-mediated EMT was detected. Results: Maternal exposure to DBP promoted the development of EMT in the offspring; Compared with the control group, DBP exposure during pregnancy led to a significant increase in the E-cadherin index of the offspring, IHC staining intensity was 3 times higher than the control group. Also, N-cadherin index decreased significantly, IHC staining intensity was about 20% of the control group (P<0.05). DBP promoted Snail1 expression and interference on Snail1 function leaded to inhibition on DBP-induced EMT (about 20% of the control group). DBP exposure was associated with high expression of TGF-β1 in the kidney, and TGF-β1 receptor inhibitor was able to inhibit the activation of Snail1 by DBP(P<0.05). Accumulation of reactive oxygen species (ROS) induced by DBP promoted the expression of TGF-β1 in renal tubular cells. Conclusion: Exposure to DBP during pregnancy leads to accumulation of ROS in the kidney and high expression of TGF-β1, which in turn promotes Snail1-mediated EMT in renal tubular epithelial cells.
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