油脂煎炸过程中TBBQ生成规律、危害调控及其细胞毒性的分子机制研究

项目拟采用细胞毒理学、分子生物学、质谱技术、脂类化学、氧化动力学、色谱技术等，针对食用油煎炸过程中叔丁基对苯二酚（TBHQ）氧化产物鉴定、主要代谢产物叔丁基对苯醌（TBBQ）的生成途径、蓄积规律及环境参数的影响开展研究，开发煎炸油中TBBQ的生成调控及选择性脱除技术，为指导建立安全、科学的煎炸工艺规范提供理论支撑。同时评估TBBQ在煎炸油、煎炸食品中的残留量及人体曝露量，解析TBBQ的细胞毒性及量效关系。重点解明TBBQ诱导细胞毒性的细胞凋亡机制，结合转录组测序技术筛选差异表达基因，并对差异表达基因GO富集和KEGG通路分析，解析TBBQ诱导细胞毒性的信号通路和关键作用靶点，并采用实时荧光定量PCR和免疫蛋白印迹(Western-blot)法验证。从分子水平上揭示TBBQ的细胞毒性机制，为煎炸食品中风险因子TBBQ的安全评估提供基础数据。

Studies on safety evaluation and control on tert-butylhydroquinone (TBBQ), the main oxidation products of TBHQ during frying with edible oil, would be carried out by means of multidisciplinary technology. The oxidated products was identified by GC/MS, and the main formation pathway of TBBQ was elucilated. The rules on TBBQ accumulation in fried oil and fried food, its environmental parameters effects and selectively removal methods would be explored. This will underlie the theoretical foundation for the development of safe and scientific frying process. Meanwhile, the residual level of TBBQ in fried oil and fried food and its exposure level to human were evaluated to confirm the cytotoxicity and dose-effect relationship of TBBQ. The key points was focused on the elucidation of cytotoxiology, to clarify which of apoptosis, inflammatory and autophagy is responsive for cytotoxicity of TBBQ. To disclose the signal pathway and key targets genes functioned by TBBQ, the transcriptome sequencing (RNA-Seq) technology was used to screen the differentially expressed genes, and GO enrichment and KEGG pathway were analyzed. The expression of key genes and its products were verified by using quantitative real-time reverse-transcriptase-polymerase chain reaction (qRT-PCR) and western blot. The clarification of the cytotoxic mechanism of TBBQ at the molecular level would provide fundermental data for the safety assessment of TBBQ in Fried food. The research is important academicly, and would be favorable for improvement of safety in fried food.