| 摘要: |
| 目的 探讨特异性上调肝细胞中肝细胞核因子1α（HNF1α）表达对四氯化碳（CCl4）诱导的小鼠肝纤维化的影响。方法 取18只C57/B6小鼠随机分为正常组、AAV8-TBG-Ctrl组和AAV8-TBG-HNF1α组，每组6只。应用CCl4诱导制备小鼠肝纤维化模型，造模后AAV8-TBG-HNF1α组小鼠通过尾静脉注射带有甲状腺素结合球蛋白（TBG）启动子驱动表达HNF1α的腺相关病毒AAV8-TBG-HNF1α特异性上调肝细胞中HNF1α的表达，AAV8-TBG-Ctrl组小鼠注射对照病毒AAV8-TBG。采用免疫组织化学法及qPCR检测各组小鼠HNF1α的表达，苏木素-伊红（H-E）染色、天狼猩红染色检测肝组织的病理改变及胶原沉积，免疫组化法检测α-平滑肌肌动蛋白（α-SMA）的表达，并对Ⅰ型胶原（COL1A1）及α-SMA的表达进行软件定量分析；qPCR法检测小鼠肝组织中纤维化相关基因（α-SMA和COL1A1）、上皮指标（E-cadherin和Plakoglobin）及间质指标（Vimentin，Slug和Twist1）的mRNA水平；免疫组化法及TUNEL法检测上调HNF1α对纤维化肝脏中细胞增殖、凋亡的影响。结果 与正常组小鼠相比，CCl4造模可促进小鼠肝脏胶原沉积及α-SMA的表达，且在肝纤维化发生过程中HNF1α的表达下降（P<0.01）；与AAV8-TBG-Ctrl组相比，应用AAV8-TBG-HNF1α特异性上调肝细胞HNF1α的表达可抑制纤维化肝脏中的COL1A1及α-SMA的水平（P<0.01）。上调肝细胞HNF1α表达对纤维化肝脏中E-cadherin、Vimentin等上皮间质转换指标的表达无明显影响（P>0.05），也不影响肝细胞的增殖与凋亡（P>0.05）。结论 特异性上调肝细胞HNF1α表达可显著改善CCl4诱导的小鼠肝纤维化。 |
| 关键词: 肝细胞核因子1α 肝细胞 肝纤维化 胶原Ⅰ型 |
| DOI：10.16781/j.0258-879x.2017.09.0000 |
| 投稿时间：2017-05-04修订日期：2017-06-29 |
| 基金项目:国家自然科学基金重点项目（81230011）. |
|
| Overexpression of hepatocyte nuclear factor 1α mediated by adeno-associated virus attenuates carbon tetrachloride-induced hepatic fibrosis in mice |
| YAO Li-jia,DENG Xing,DING Chen-hong,FENG Ren-xin,XIE Wei-fen* |
(Department of Gastroenterology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*Corresponding author) |
| Abstract: |
| To explore the influence of the hepatocyte-specific up-regulation of hepatocyte nuclear factor 1α (HNF1α) on mouse hepatic fibrosis induced by carbon tetrachloride (CCl4). Methods Eighteen C57/B6 male mice were randomly divided into normal group, AAV8-TBG-Ctrl group and AAV8-TBG-HNF1α group, with 6 mice in each group. Mice in the AAV8-TBG-Ctrl and AAV8-TBG-HNF1α groups were intraperitoneally injected with CCl4 to establish the hepatic fibrosis mouse model, and then the mice in the AAV8-TBG-HNF1α group were injected with AAV8-TBG-HNF1α carrying HNF1α gene under the control of the thyroid-binding globulin (TBG) promoter to specifically up-regulate expression of HNF1α in hepatocytes, while the mice in the AAV8-TBG-Ctrl group were injected with control vector AAV8-TBG. The expression of HNF1α was determined by immunohistochemistry and qPCR. The pathological changes and collagen deposition of liver tissues were detected by hematoxylin-eosin (H-E) staining and sirius red staining, respectively. Immunohistochemistry method was used to detect α-smooth muscle actin (α-SMA), and the expression of fibrosis related genes (typeⅠcollagen α1 chain[COL1A1], α-SMA), epithelial related genes (E-cadherin, Plakoglobin) and mesenchymal related genes (Vimentin, Slug and Twist1) in liver tissues were analyzed by qPCR. The cell proliferation and apoptosis in fibrotic livers were detected by immunohistochemistry and TdT-mediated dUTP Nick-End Labeling (TUNEL) method, respectively. Results Compared with the normal mice, CCl4 promoted collagen deposition and the expression of α-SMA in livers, and the expression of HNF1α was significantly decreased (P<0.01) in the process of hepatic fibrosis. Compared with the AAV8-TBG-Ctrl group, the expression of COL1A1 and α-SMA in the fibrotic livers in the AAV8-TBG-HNF1α group was significantly decreased (P<0.01). There was no significant difference in the expression of E-cadherin, Vimentin or other Epithelial-mesenchymal transition related genes, or in the cell proliferation and apoptosis between the AAV8-TBG-Ctrl and AAV8-TBG-HNF1α groups (P>0.05). Conclusion Hepatocyte-specific up-regulation of HNF1α significantly improves CCl4-induced liver fibrosis in mice. |
| Key words: hepatocyte nuclear factor 1α hepatocytes liver fibrosis collagen type Ⅰ |
