【摘要】 目的:观察CLC氯通道/转运蛋白家族(CLC3)氯离子通道蛋白在不同细胞系中的表达,明确CLC3蛋白在人肝实质细胞内的表达及亚细胞定位,为阐明CLC3在非酒精性脂肪肝中的作用及可能机制提供理论依据。方法:培养不同来源的细胞株,包括小鼠和人的肝实质细胞株,Western blot方法检测CLC3蛋白在肝细胞的表达;免疫荧光和蔗糖密度梯度离心方法检测CIC3在肝细胞内的表达定位。结果:在不同来源的细胞株中均见CLC3蛋白的表达且肝实质细胞有较大表达量;梯度密度离心分离肝细胞亚细胞组分,发现内吞体蛋白和CLC3蛋白峰值同时出现;免疫荧光发现CLC3蛋白与内吞体双染。结论:CLC3蛋白在肝实质细胞相对高表达并主要定位于内吞体。更多还原

【Abstract】 Objective To observe the expression of CLC3 chloride channel protein in different cell lines , to define the subcellular localization and expression of CLC3 chloride channel protein in hepatocytes of Homo sapiens, and to provide the theory for investigating the physiopathological role of CLC3 chloride channel in the development of non-alcoholic fatty liver disease (NAFLD). Methods Hepatic cell lines from human and from mouse were cultured in vitro. Western blot analysis was used to detect the expression of CLC3 chloride channel protein in thses cell lines. Subcellular localization of CLC3 chloride channel protein in hepatocytes was detected by sucrose density gradient centrifugation and immunofluorescence. Results CLC3 chloride channel protein was expressed stably in cell lines from human and from mouse, especially in hepatocytes. The peak expressions of CLC3 chloride channel protein and endosomes were presented at the same gradient. Conclusion CLC3 chloride channel protein was highly expressed in hepatocytes, and was localized principally in endosomes in hepatocytes.更多还原