Department of Pediatric Surgery, Children's Hospital of Fudan University, and Key Laboratory of Neonatal Disease, Ministry of Health, Shanghai, China.
J Pediatr Gastroenterol Nutr. 2013 Dec;57(6):713-7. doi: 10.1097/MPG.0b013e3182a7e1da.
Biliary atresia (BA) is a devastating pediatric cholestatic liver disease. Increasing evidence indicates that nuclear factor (NF)-κB signaling plays a key role in the pathogenesis of BA. Leucine zipper downregulated in cancer 1 (LDOC1) may control the expression of NF-κB. The aim of this study was to evaluate the relation between LDOC1 and inflammation/apoptosis mediated by NF-κB in the human intrahepatic biliary epithelial cells (HIBECs).
HIBECs were divided into 3 treatment groups: control, mock transfection group, and LDOC1 transfection. Immunofluorescence, reverse transcription polymerase chain reaction, Western blot, and flow cytometry analysis were used to investigate the effectiveness of LDOC1-transfected HIBECs and the expression of NF-κB. Apoptosis was detected by Hochest/ propidium iodide staining. Interleukin (IL)-2 and tumor necrosis factor (TNF)-α levels were evaluated by enzyme-linked immunosorbent assay.
The expression of NF-κB was higher in the LDOC1-transfected group when compared with the control and mock-transfected groups as evaluated by immunofluorescence, reverese transcription polymerase chain reaction, and Western blot analysis. The rate of apoptosis was significantly lower in the LDOC1-transfected group when compared with the control and mock-transfected groups. The levels of IL-2 and TNF-α were significantly higher in the LDOC1-transfected group when compared with the control and mock-transfected groups.
Upregulation of LDOC1 in HIBEC increases the expression of NF-κB, which may promote the activation of IL-2 and TNF-α secretion and inhibit cell apoptosis.
胆道闭锁（BA）是一种毁灭性的小儿胆汁淤积性肝病。越来越多的证据表明，核因子（NF）-κB 信号通路在 BA 的发病机制中起关键作用。亮氨酸拉链下调癌 1（LDOC1）可能控制 NF-κB 的表达。本研究旨在评估 LDOC1 与 NF-κB 介导的人肝内胆管上皮细胞（HIBEC）炎症/凋亡之间的关系。
将 HIBEC 分为 3 组：对照组、mock 转染组和 LDOC1 转染组。采用免疫荧光、逆转录聚合酶链反应、Western blot 和流式细胞术分析，研究 LDOC1 转染 HIBEC 的效果和 NF-κB 的表达。采用 Hochest/碘化丙啶染色检测细胞凋亡。采用酶联免疫吸附试验检测白细胞介素（IL）-2 和肿瘤坏死因子（TNF）-α 水平。
免疫荧光、逆转录聚合酶链反应和 Western blot 分析显示，与对照组和 mock 转染组相比，LDOC1 转染组 NF-κB 的表达更高。与对照组和 mock 转染组相比，LDOC1 转染组细胞凋亡率显著降低。与对照组和 mock 转染组相比，LDOC1 转染组 IL-2 和 TNF-α 水平显著升高。
HIBEC 中 LDOC1 的上调增加了 NF-κB 的表达，这可能促进了 IL-2 和 TNF-α 的分泌激活和细胞凋亡抑制。
