Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, 200080, China.
Pathology Center, Shanghai General Hospital/Faculty of Basic Medicine, Shanghai Jiao Tong University, School of Medicine, Shanghai, 200080, China.
Ecotoxicol Environ Saf. 2020 Dec 1;205:111154. doi: 10.1016/j.ecoenv.2020.111154. Epub 2020 Aug 15.
The study focused on the toxicological effect of Di-n-butyl phthalate (DBP) on the expression of Phosphorylated signal transducer and activator of transcription 1 (pSTAT1) -regulated Forkhead box protein M1 (FoxM1), which might provide a new understanding of gestational diabetes mellitus (GDM) development and a potential target for treatment. Streptozotocin (STZ) (40 mg/kg) was introduced in maternal rats by intraperitoneal injection on gestation day 0 (GD 0) in the STZ and STZ + DBP groups. DBP was introduced in maternal rats by oral feeding in the STZ + DBP group over the following 3 days (750 mg/kg/day). The changes in fasting blood glucose level in rats were detected on GD 1 and GD 5. The insulin levels in maternal rats and PIBCs were measured on GD 18. The Oral Glucose Tolerance Test (OGTT) test was performed on GD 18 to check the stability of the GDM model. The primary islet β cells (PIBCs) were established for in vitro experiments. We examined the FoxM1 and pSTAT1 expression in pancreas by immunohistochemistry. Real-time PCR and Western blot were used to detect the pSTAR1 and FoxM1 protein and mRNA gene expression levels in PIBCs. Cell Counting Kit-8 (CCK-8) and flow cytometric analysis was used to test the viability and apoptosis of cells. The results showed that the STZ + DBP group had higher glucose and lower insulin secretion levels than the other groups by both fasting test and OGTT. FoxM1 was significantly suppressed while pSTAT1 was highly expressed after DBP exposure. FoxM1 could be regulated by pSTAT1. DBP can influence the progression of GDM through its toxicological effect, which significantly increases the expression of pSTAT1 and suppresses FoxM1, causing a decline in β cell viability.
本研究聚焦于邻苯二甲酸二正丁酯（DBP）对磷酸化信号转导和转录激活因子 1（pSTAT1）调节的叉头框蛋白 M1（FoxM1）表达的毒理学效应，这可能为妊娠期糖尿病（GDM）的发展提供新的认识，并为治疗提供新的靶点。链脲佐菌素（STZ）（40mg/kg）于妊娠 0 天（GD 0）经腹腔注射引入母鼠，STZ 和 STZ+DBP 组中。DBP 通过灌胃在 STZ+DBP 组中连续 3 天（750mg/kg/天）给药。在 GD 1 和 GD 5 检测大鼠空腹血糖水平的变化。在 GD 18 测量母鼠和 PIBC 中的胰岛素水平。在 GD 18 进行口服葡萄糖耐量试验（OGTT）检查 GDM 模型的稳定性。进行体外实验，建立原代胰岛β细胞（PIBC）。通过免疫组织化学检测胰腺中的 FoxM1 和 pSTAT1 表达。实时 PCR 和 Western blot 用于检测 PIBC 中 pSTAR1 和 FoxM1 蛋白和 mRNA 基因表达水平。细胞计数试剂盒-8（CCK-8）和流式细胞术分析用于检测细胞活力和凋亡。结果表明，STZ+DBP 组的空腹和 OGTT 试验血糖均高于其他组，胰岛素分泌水平较低。DBP 暴露后 FoxM1 明显受抑制，而 pSTAT1 高表达。FoxM1 可受 pSTAT1 调节。DBP 通过其毒理学效应影响 GDM 的进展，显著增加 pSTAT1 的表达并抑制 FoxM1，导致β细胞活力下降。
