文章摘要
草鱼体内溶血对肝脏氧化损伤的机制研究
Study on the mechanism of liver oxidative damage induced by hemolysis in grass carp
投稿时间:2021-02-26  修订日期:2021-03-19
DOI:
中文关键词: 草鱼  血红蛋白  氧化损伤  溶血
英文关键词: Ctenopharyngodon idellus  oxidative damage  hemoglobin  hemolysis
基金项目:
作者单位邮编
秦真东 仲恺农业工程学院动物科技学院 510225
卢志杰 仲恺农业工程学院动物科技学院 
杨敏璇 仲恺农业工程学院动物科技学院 
黄垚 仲恺农业工程学院动物科技学院 
詹凡玢 仲恺农业工程学院动物科技学院 
李亚男 仲恺农业工程学院动物科技学院 
施斐 仲恺农业工程学院动物科技学院 
陈少君 仲恺农业工程学院动物科技学院 
李军 仲恺农业工程学院动物科技学院 
林蠡 仲恺农业工程学院动物科技学院 510225
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中文摘要:
      多种细菌或者病毒性病通常导致鱼体大量出血,而释放大量细胞毒性的血红蛋白进入组织中,目前有关其对鱼体组织的损伤机制尚未有详细研究。本研究以草鱼为研究对象,通过模拟体内出血,系统研究出血对肝脏的损伤机制。本研究首先在体内注射血红蛋白,通过HE染色发现注射血红蛋白组的肝脏组织中出现明显坏死细胞增多,普鲁士蓝染色揭示肝脏中存在大量的铁沉积,进一步的荧光定量PCR检测结果显示注射的血红蛋白激活了铁代谢相关基因的表达。为进一步探究体内出血对肝脏的影响,本研究通过体内注射苯肼模拟体内出血,HE和普鲁士蓝染色结果表明大量的溶血导致肝脏细胞坏死和铁的沉积增加,通过检测肝脏中血红蛋白和铁含量发现,苯肼组中肝脏组织中的血红蛋白和铁含量随着时间延长而显著增加,并且铁含量的增加同时激活了肝脏组织中铁代谢相关基因的表达。其次,我们检测了注射苯肼后肝脏组织中炎症因子的表达情况,荧光定量PCR结果显示,高剂量的血红蛋白激活了多种细胞因子的表达,如促炎因子TNF-α,IL-1β和IL-6,抑炎因子IL-10以及趋化因子IL-4和IL-8等。为进一步探究高氧化活性的血红蛋白对肝脏组织的氧化损伤作用,我们检测了肝脏组织中丙二醛(MDA),脂质过氧化物(LPO)以及也β-半乳糖苷酶的含量情况,检测结果表明,鱼体内的出血显著增加了肝脏氧化损伤作用,同时,荧光定量PCR和酶活检测结果揭示,血红蛋白的氧化损伤作用促进了肝脏组织细胞凋亡的发生。最后,我们也检测了肝脏组织中抗氧化酶的表达情况,结果显示,体内的出血显著激活了肝脏组织中的抗氧化系统。综上所述,本研究表明,鱼体内大量出血释放的高氧化活性的血红蛋白显著激活了肝脏组织中的炎症和氧化损伤的发生,促进了肝脏组织中的细胞凋亡,同时也激活了机体内的抗氧化系统。
英文摘要:
      A variety of bacterial or viral diseases usually cause massive hemolysis in fish, and release a large amount of cytotoxic hemoglobin into the tissues. At present, the damage mechanism of cytotoxic hemoglobin to fish tissues has not been well studied. In this study, grass carp was used as the research model to systematically explore the damage mechanism of high oxidation hemoglobin to liver. Firstly, the hemoglobin was injected into the grass carp, HE staining assay results showed that injection of hemoglobin caused the dead cells increased obviously in liver, and Prussian blue staining revealed that more iron deposited in liver, the fluorescence quantitative PCR (qRT-PCR) detection results showed that the injection of hemoglobin activated the expression of iron metabolism related genes. In order to further explore the damage of hemolysis to the liver, we injected phenylhydrazine into grass carp, HE and Prussian blue staining results showed that the hemolysis caused the liver cell necrosis and iron deposits increased, the content of hemoglobin and iron in the liver all significantly increased with time extended. Furthermore, the increase of iron also activated the expression of iron metabolism related genes. Followed, we detected the expression of inflammatory cytokines in liver after the injection of phenylhydrazine, qRT-PCR results showed that high dose of hemoglobin activated the expression of various cytokines, such as pro-inflammatory cytokines TNF-α, IL-1β and IL-6, anti-inflammatory cytokines IL-10, and chemokines IL-4 and IL-8. In order to further explore the oxidation damage of hemoglobin in liver, we tested the content of malondialdehyde (MDA) and lipid peroxide (LPO) and β - galactose glucoside enzyme in liver, the results showed that the hemolysis significantly increased liver oxidative damage effect, at the same time, the qRT-PCR and enzyme activity test results revealed that the oxidative damage of hemolysis promoted the occurrence of liver cell apoptosis. Finally, we also examined the expression of antioxidant enzymes in liver, and the results showed that in vivo hemolysis significantly activated the antioxidant system in liver. In conclusion, the present study revealed that massive hemorrhage in fish released the high oxidative activity of hemoglobin which activated the occurrence of inflammation, oxidative damage and apoptosis in liver, and simultaneously up-regulated the expression of antioxidant system.
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