Aquaporin-9 downregulation prevents steatosis in oleic acid-induced non-alcoholic fatty liver disease cell models
Aquaporin-9 downregulation prevents steatosis in oleic acid-induced non-alcoholic fatty liver disease cell models
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Aquaporin-9 下调可预防油酸诱导的非酒精性脂肪肝细胞模型中的脂肪变性
DOI：
10.3892/ijmm.2013.1502
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发表时间：
2013-11-01
影响因子：
5.4
通讯作者：
Jiang, Zheng
中科院分区：
文献类型：
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作者：
Wang, Chuan;Lv, Zi-Lan;Jiang, Zheng
水通道蛋白-9（AQP 9）是一种水甘油通道蛋白，充当脂肪甘油通道。然而，AQP 9在非酒精性脂肪性肝病（NAFLD）脂肪变性中的作用尚未完全阐明。本研究采用RT-PCR方法从LO 2细胞中获得AQP 9基因编码序列，并将其克隆到pEGFP-N1载体中。将靶向AQP 9基因的短发夹RNA（shRNA）插入pGenesil-1载体中。重组质粒通过酶切和序列分析确认，并转染到油酸诱导的NAFLD的细胞模型（来自LO 2细胞）中。我们的结果表明，AQP 9重组质粒可以有效地表达在细胞模型的NAFLD。此外，与对照组相比，AQP 9过表达显著增加细胞内脂质含量，甘油三酯（TG），游离脂肪酸（FFA）和甘油水平;然而，AQP 9的沉默产生相反的效果。总之，成功构建了用于诱导AQP 9过表达和沉默AQP 9表达的重组质粒。AQP 9过表达加重了脂肪变性的程度;然而，AQP 9的沉默减轻了这些作用。基于这些数据，我们认为AQP 9可能作为一种新的分子靶点，用于NAFLD的治疗干预。
Aquaporin-9 (AQP9) is an aquaglyceroporin that acts as the adipose glycerol channel. However, the role of AQP9 in steatosis in non-alcoholic fatty liver disease (NAFLD) has not yet been fully elucidated. In the present study, the coding sequence of the AQP9 gene was obtained from LO2 cells by RT-PCR, and cloned into the pEGFP-N1 vector. Short hairpin RNA (shRNA) targeting the AQP9 gene was inserted into the pGenesil-1 vector. Recombinant plasmids were confirmed by enzyme digestion and sequence analysis, and transfected into cell models (derived from LO2 cells) of oleic acid-induced NAFLD. Our results demonstrated that AQP9 recombinant plasmids can be effectively expressed in cell models of NAFLD. Furthermore, in comparison with the control group, AQP9 overexpression significantly increased intracellular lipid content, triglyceride (TG), free fatty acid (FFA) and glycerol levels; however, the silencing of AQP9 exerted the opposite effects. Taken together, recombinant plasmids used to induce AQP9 overexpression and to silence AQP9 expression were successfully constructed. AQP9 overexpression aggravated the degree of steatosis; however, the silencing of AQP9 alleviated these effects. Based on these data, we suggest that AQP9 may serve as a novel molecular target for therapeutic intervention in NAFLD.