School of Biotechnology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India.
School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India.
Sci Rep. 2018 Jan 22;8(1):1310. doi: 10.1038/s41598-018-19299-w.
This study deals with the isolation and purification of an important variant of microcystins namely microcystin-RR (MCYST-RR) from Microcystis aeruginosa and reports its effects on mice liver protein profile and cellular functions. Protein profiling by 2-dimensional gel electrophoresis revealed changes in the number and accumulation of protein spots in liver of mice treated with different concentrations of MCYST-RR. Untreated (control) mice liver showed 368 protein spots while the number was 355, 348 and 332 in liver of mice treated with 200, 300 and 400 µg kg body wt of MCYST-RR respectively. Altogether 102, 97, and 92 spots were differentially up-accumulated and 93, 91, and 87 spots were down- accumulated respectively with the treatment of 200, 300, 400 µg kg body wt. Eighteen differentially accumulated proteins present in all the four conditions were identified by MALDI-TOF MS. Of these eighteen proteins, 12 appeared to be involved in apoptosis/toxicological manifestations. Pathway analysis by Reactome and PANTHER database also mapped the identified proteins to programmed cell death/apoptosis clade. That MCYST-RR induces apoptosis in liver tissues was also confirmed by DNA fragmentation assay. Results of this study elucidate the proteomic basis for the hepatotoxicity of MCYST-RR which is otherwise poorly understood till date.
本研究从铜绿微囊藻中分离和纯化了一种重要的微囊藻毒素变体，即微囊藻毒素-RR（MCYST-RR），并报告了其对小鼠肝脏蛋白质谱和细胞功能的影响。通过二维凝胶电泳进行蛋白质谱分析显示，用不同浓度的 MCYST-RR 处理的小鼠肝脏中蛋白质斑点的数量和积累发生了变化。未经处理（对照）的小鼠肝脏显示出 368 个蛋白质斑点，而用 200、300 和 400μg/kg 体重的 MCYST-RR 处理的小鼠肝脏中蛋白质斑点的数量分别为 355、348 和 332。总共分别有 102、97 和 92 个斑点被上调积累，93、91 和 87 个斑点被下调积累，用 200、300、400μg/kg 体重进行处理。通过 MALDI-TOF MS 鉴定了在所有四种条件下存在的 18 个差异积累的蛋白质。这 18 种蛋白质中有 12 种似乎与细胞凋亡/毒性表现有关。Reactome 和 PANTHER 数据库的途径分析也将鉴定的蛋白质映射到程序性细胞死亡/凋亡簇。MCYST-RR 诱导肝脏组织细胞凋亡的结果也通过 DNA 片段化测定得到了证实。本研究结果阐明了 MCYST-RR 肝毒性的蛋白质组学基础，而这在迄今为止尚未得到很好的理解。
