内质网分子伴侣PDI对肝细胞脂肪变性的影响.docVIP

  内质网分子伴侣 PDI 对肝细胞脂肪变性的 影响# * 5 10 15 20 25 30 35 40  摘要：目的：探讨内质网分子伴侣蛋白质二硫键异构酶 PDI 对肝细胞脂肪变性的影响。方 法：用高脂培养基处理人正常肝细胞 L02 细胞，建立肝细胞脂肪变性的细胞模型；用 Western blot 检测正常组和模型组细胞 PDI 的表达水平；利用脂质体 Lipofectamine2000 将针对靶基 因 PDI 合成的四条特异性 siRNA 片段及阴性对照 siRNA 转染 L02 细胞，Western blot 检测 PDI 基因表达变化，进行有效干扰片段的筛选；用高脂培养基处理 PDI 表达抑制的 L02 细 胞，用油红染色观察细胞脂肪变的程度。结果：与正常对照组相比较，脂变组细胞 PDI 表 达水平显著升高（P0.05）；用 siRNA 抑制 PDI 的表达，与阴性对照组相比，肝细胞脂肪 变性的程度明显减轻。结论：PDI 表达抑制能减轻高脂培养基诱导的肝细胞脂肪变性。 关键词：内科学；蛋白质二硫键异构酶；肝脏；脂肪变性 中图分类号：R575.5 Effect of protein disulfide isomerase on hepatocyte steatosis ZHANG Xuequn, PAN Yue, HUANG Jierong, YU Chaohui, LI Youming (Digestive Department, the First Affiliated Hospital, Zhejiang University,310001) Abstract: Objective: To investigae the potential role of ER chaperone protein disulfide isomerase (PDI) on the hepatocyte steatosis. Methods: The human hepatocyte cell line L02 cells were exposed to a mixture of FFAs (oleate and palmitate); The protein level of PDI was detected by Western blot analysis; siRNA targeting PDI gene and negative siRNA were transfected into L02 cells using Lipofectamine 2000. The interfering efficiency of the siRNAs was assessed by Western blotting; L02 cells with PDI gene knockdown were cultred in FFAs for 24 hours. The cellular lipid accumulation was assessed by Oil Red O staining. Results: The cellular expression level of PDI was significantly increased after FFAs treatment. The siRNA-mediated knockdown of PDI in L02 cells demonstrated decreased cellular lipid accumulation induced by FFA overload. Conclusion: PDI down-regulation decreased cellular lipid accumulation induced by FFA overload. Key words: internal medicine; protein disulfide isomerase; hepatic; steatosis 0 引言 非酒精性脂肪性肝病（nonalcoholic fatty liver disease, NAFLD）是指非酒精原因所致的 以肝细胞脂肪变性为主要特征的临床病理综合征[1]。近年来随着生活方式的改变，NAFLD 的患病率在逐年增高，已经成为导致慢性肝病的重要原因之一，构成日益严重的社会健康问 题[2,3]。NAFLD 可以从单纯性脂肪肝向脂肪性肝炎、肝纤维化、肝硬化等终末期肝病进展[4,5]。 其确切的分子机制尚未完全明确。 蛋白质二硫键异构酶（protein disulfide isomerase, PDI）是一种定位于内质网管腔的多功 能分子伴侣，它同时具有分子伴侣和氧