氢醌刺激后人肝细胞蛋白质谱表达的变化

目的研究氧醌(HQ)刺激后L-02型人肝细胞蛋白质谱的表达变化。方法传代培养的L-02型肝细胞分为空白对照组和染毒处理组。处理组经HQ(1×10-4mol／L)染毒培养24 h,对照组不做处理。提取蛋白同时进行双向凝胶电泳,图谱经PDQuest软件分析寻找差异点。实验独立进行3次,每次处理组和对照组各设3块胶作为组内重复,切取重复性较好的差异点做MALDI-TOF质谱鉴定,得到PMF通过Mascot在美国国家生物信息检索中心(NCBI)人类蛋白质数据库搜索。结果每块胶检测到约1 000个蛋白点,对照组和处理组凝胶匹配率组内达90％以上,组间80％以上,3次实验各发现17、18、24个蛋白点表达改变,变化倍数≥2。重复性较好的4个蛋白点经MALDI-TOF质谱鉴定为Rho GDP解离抑制蛋白、6-磷酸葡萄糖内酯酶、erbB3结合蛋白EBP1和核纤层蛋白,其功能与信号转导、细胞骨架及能量代谢有关。结论1×10-4mol／L的HQ处理24 h可引起L-02型人肝细胞蛋白质谱表达改变。

Differential proteomic expression in human liver cells stimulated by hydroquinone

OBJECTIVE: To explore the differential proteomic expression in human liver cells L-02 after exposure to HQ. METHODS: Subcultured L-02 cells were treated by HQ for 24 h at a 1 x 10(-4) mol/L concentration and a blank group was set as the control. Immediately after the treatment, total cellular proteins were extracted and separated by 2-DE, and the images were analyzed by PDQuest software. The experiment was totally repeated 3 times with 3 repetitions for each group every time. The well repeated spots were identified by MALDI-TOF MS and then searched in NCBI human protein database with Mascot. RESULTS: About 1,000 spots per gel were found. Compared with the control group, 17, 18 and 24 spots were significantly altered in 3 separate experiments. The 4 well repeated spots were identified by MALDI-TOF MS as Rho GDP dissociation inhibitor GDI alpha, 6-phosphogluconolactonase, erbB3 binding protein EBP1 and lamin A/C, isoform 1 precursor. They were involved in cell skeleton, signal transduction and energy metabolization in functional classification. CONCLUSION: Hydroquinone can change the protein expression in liver cells, which provides clues for exploring the toxic mechanism.