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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。
Objective To study the changes of protein profiles of L-02 human hepatocytes stimulated with oxyquinone (HQ). Methods L-02 hepatocytes subcultured were divided into blank control group and exposure group. The treatment group was exposed to HQ (1 × 10-4mol / L) for 24 hours and the control group was not treated. Extraction of protein at the same time two-dimensional gel electrophoresis, PDQuest software analysis of the map looking for differences. The experiments were carried out 3 times independently, with 3 gums in each treatment group and control group as in-group repeats. MALDI-TOF mass spectrometry was used to identify the differences with good reproducibility. PMF was screened by Mascot at the National Center for Biological Information Retrieval NCBI) Human Protein Database Search. The results showed that about 1 000 protein spots were detected per gel, and the gel matching rate of the control group and the treatment group reached more than 90%, with more than 80% between the groups. In the three experiments, 17, 18, and 24 protein spots were changed , Change multiple ≥2. The four repeats were identified as Rho GDP dissociation inhibitor, 6-phosphoglucuronidase, erbB3-binding protein EBP1 and lamin, which were identified by MALDI-TOF mass spectrometry. Their function and signal transduction, cytoskeleton And energy metabolism. Conclusions Treatment of HQ with 1 × 10-4 mol / L for 24 h caused a change in protein profile of L-02 human hepatocytes.