微囊藻毒素-LR对中华绒螯蟹肝胰腺的毒性及其机理的研究

The liver is the main target organ of microcystin. Microcystin-LR（MC-LR）can cause liver tissue damage in mammals, inhibits the activity of protein phosphatase 2A（PP2A）and thus, actives the downstream MAPK pathway. However, the toxicity and its mechanism of MC-LR to the hepatopancreas of Eriocheir sinensis is not reported. This project is based on the hepatopancreas of E. sinensis. Firstly, clarifies the toxicity of MC-LR to the hepatopancreas of E. sinensis by the study of oxidative damage, antioxidation system, histological pathology, and apoptosis after exposure to MC-LR. In order to explore the relationship between cytoskeleton recombination and apoptosis induced by MC-LR, and then investigate the changes in the morphology, distribution and the phosphorylation level of the cytoskeletal proteins in the hepatocytes using of immunofluorescence technology and western blotting technology when the activity of PP2A is inhibited most obviously. On this basis, further studies on the regulating effects of PP2A and MAPK signaling pathway on the phosphorylation of cytoskeletal proteins will be conducted, to thoroughly clarify its toxicity in hepatopancreas of E. sinensis and reveal the molecular mechanism of hepatopancreas tissue damage.

肝脏是微囊藻毒素主要的靶器官。微囊藻毒素-LR（MC-LR）引起哺乳动物肝脏损伤，抑制PP2A酶活性，激活MAPK通路。然而，MC-LR对中华绒螯蟹肝胰腺的毒性仍未见报道，对肝胰腺毒性的机理仍不明确。本项目以中华绒螯蟹肝胰腺为研究对象，首先对在MC-LR暴露下肝胰腺氧化损伤、抗氧化系统、组织病理学和肝胰腺细胞凋亡情况进行研究，初步揭示MC-LR对中华绒螯蟹肝胰腺的毒性；其次，利用免疫荧光技术和免疫印迹技术进一步研究PP2A酶活性被抑制时细胞骨架形态、分布以及细胞骨架相关蛋白磷酸化水平的变化，进一步探讨MC-LR诱导细胞骨架重组与细胞凋亡的关系；最后，研究PP2A和MAPK信号通路对细胞骨架相关蛋白磷酸化的调控作用，阐明MC-LR对中华绒螯蟹肝胰腺及细胞的毒性，初步揭示肝胰腺组织损伤及分子机理。

肝脏是微囊藻毒素主要的靶器官。微囊藻毒素-LR（MC-LR）引起哺乳动物肝脏损伤，抑制PP2A酶活性，激活MAPK通路。然而，MC-LR对中华绒螯蟹肝胰腺的毒性仍未见报道，对肝胰腺毒性的机理仍不明确。本项目以中华绒螯蟹肝胰腺为研究对象，首先对在MC-LR暴露下造成中华绒螯蟹肝胰腺氧化应激ROS和LPO水平提高、抗氧化相关酶活和基因表达发生变化、肝胰腺组织形态和肝胰腺细胞亚细胞结构发生改变，肝胰腺细胞凋亡水平提高，初步揭示了MC-LR对中华绒螯蟹肝胰腺的毒性；其次，利用免疫荧光技术和免疫印迹技术进一步研究PP2A酶活性被抑制时发现果蝇S2细胞骨架形态、分布发生变化，中华绒螯蟹肝胰腺细胞骨架相关TAU蛋白磷酸化水平表达提高，进一步探讨了MC-LR诱导细胞骨架重组与细胞凋亡的关系；最后，研究了PP2A和MAPK信号通路对细胞骨架相关蛋白磷酸化的调控作用，结果显示p38、JNK和ERK1/2蛋白磷酸化水平提高，MAPK通路被激活，进一步阐明了MC-LR对中华绒螯蟹肝胰腺及细胞的毒性，初步揭示了肝胰腺组织损伤及细胞凋亡的分子机理。