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

作　　者：赵圣[1] 李登[1] 朱依萍 贝晓宇 蒋君涛[1] 韩邦旻[1] ZHAO Sheng;LI Deng;ZHU Yi-ping;BEI Xiao-yu;JIANG Jun-tao;HAN Bang-min(Department of Urology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200080,China;Institute of clinical transformation,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200080,China)

机构地区：[1]上海交通大学附属第一人民医院泌尿外科,上海200080 [2]上海交通大学附属第一人民医院临床转化研究院,上海200080

出　　处：《现代生物医学进展》2019年第12期2207-2213,共7页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81771564)

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。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 NRK52 E 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. Conclusions: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.

关 键 词：邻苯二甲酸二正丁酯 上皮-间充质转化 SNAIL1 肾纤维化 

分 类 号：R-33[医药卫生] R339.2