微囊藻毒素-LR对小鼠原代肝细胞线粒体功能的影响  

Mitochondrial function in microcystin-LR-induced primary mice hepatocytes

作　　者：贺云 陈艺生 任岚 尹一杰 胡桂玲 吴云丽 HE Yun;CHEN Yisheng;REN Lan;YIN Yijie;HU Guiling;WU Yunli(Key Laboratory of Gastrointestinal Cancer(Fujian Medical University),Ministry of Education,Fujian Key Laboratory of Tumor Microbiology,School of Basic Medical Sciences,Fujian Medical University,Fuzhou 350122,China;Center for Experimental Research in Clinical Medicine,Fujian Provincial Hospital,Fuzhou 350001)

机构地区：[1]福建医科大学基础医学院,消化道恶性肿瘤教育部重点实验室,福建省肿瘤微生物学重点实验室,福州350122 [2]福建省立医院临床医学实验研究中心,福州350001

出　　处：《中国实验动物学报》2021年第6期816-822,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：福建省中青年教师教育科研项目(JT180165);福建医科大学启航基金项目(2018QH1006);福建省高等学校新世纪优秀人才支持计划(2018B027)。

摘　　要：目的蓝藻水华引起的微囊藻毒素污染是世界性关注话题之一,微囊藻毒素LR(MC-LR)具有强特异性肝毒性,但其引起肝损伤的确切机制尚未完全阐明。为解决这一问题,本研究从细胞分子层面探讨MC-LR造成肝细胞线粒体功能改变的分子机制。方法提取小鼠原代肝细胞,加入梯度剂量的MC-LR(2.5~10 nmol/L)作用48 h,以未加毒素处理组为对照,检测MC-LR对线粒体功能的影响(包括ATP水平和线粒体膜电位检测),DNA损伤(包括彗星试验和8-OHdG水平检测),并分析p53抑制剂pft-α作用下线粒体功能损伤情况。结果 MC-LR造成肝细胞线粒体功能障碍,DNA损伤且p53蛋白表达水平上调;p53特异性抑制剂pft-α减轻MC-LR造成的线粒体损伤。结论在本试验条件下,MC-LR造成小鼠肝细胞线粒体功能异常的机制与其造成DNA损伤诱导p53上调有关。Objective Microcystin contamination caused by cyanobacteria blooms is a profound global concern. Microcystin-LR(MC-LR) was documented to induce potent hepatotoxicity, but the exact mechanisms have not been fully elucidated. To better understand the mechanisms, we conducted the following investigations. Methods Primary mouse hepatocytes were isolated, purified and exposed to a MC-LR concentration range of 2.5 ~ 10 nmol/L for 48 h with DMEM treatment as the negative control. Mitochondrial function(ATP levels and mitochondrial membrane potential), DNA damage(comet assay and 8-OHdG levels) and p53 expression were determined in MC-LR-treated cells and control cells. In addition, experiments were repeated to assess mitochondrial function after pft-α preconditioning. Results MC-LR led to a progressive loss in ATP production and mitochondrial membrane potential in a dose-dependent manner. Furthermore, the comprehensive DNA damage examination and upregulated p53 protein level indicated that MC-LR was associated with DNA damage in primary mouse hepatocytes. However, when p53 was blocked by PFT-α, mitochondrial damage was attenuated after MC-LR treatment. Conclusions Together, these data indicate that MCLR-induced DNA damage promoted p53 production, which was associated with mitochondrial dysfunction.

关 键 词：微囊藻毒素-LR 原代肝细胞 线粒体功能 DNA损伤 P53 

分 类 号：Q95-33[生物学—动物学]