脂多糖诱导黑龙江林蛙炎症损伤中抗氧化机制的研究  

Study on the antioxidant effect of Rana amurensis induced by LPS in inflammatory damage

作　　者：刘依铭 许冬梅 楼柏丹[1] 刘玉芬[1] 刘鹏[1] 赵文阁[1] LIU Yi-ming;XU Dong-mei;LOU Bai-dan;LIU Yu-fen;LIU Peng;ZHAO Wen-ge(Life Science and Technology,Harbin Normal University,Harbin 150025,China)

机构地区：[1]哈尔滨师范大学生命科学与技术学院,黑龙江哈尔滨150025

出　　处：《中国预防兽医学报》2023年第7期744-749,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省自然科学基金联合引导项目(LH2021C053)。

摘　　要：为探究在细菌脂多糖(LPS)诱导下,黑龙江林蛙(Rana amurensis)抗氧化酶活性的变化,以及铁蛋白重链亚基基因(Ferritin-H,Fer-H)参与林蛙机体抗氧化反应的作用机制,本研究以1 m L/只LPS溶液(1 mg/mL)腹腔注射黑龙江林蛙,构建LPS诱导的黑龙江林蛙炎症模型,诱导后各时间点(6 h、24 h、48 h、72 h和120 h)采集组织样品,采用HE染色法观察各组织病变,结果显示,LPS诱导下黑龙江林蛙的肝脏、皮肤以及肌肉组织均出现了损伤,LPS诱导的林蛙炎症模型可以用于后续实验。利用相应的活性测定试剂盒对采集LPS诱导后各时间点林蛙各组织中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)以及谷胱甘肽过氧化物酶(GSH)等抗氧化酶活性检测,同时采用荧光定量PCR检测黑龙江林蛙各组织中Fer-H基因转录水平的变化。抗氧化酶活性检测结果显示,和未加LPS的对照组相比,LPS诱导后黑龙江林蛙各组织中SOD、CAT的活性均极显著升高(P<0.01),而GSH活性极显著下降(P<0.01);荧光定量PCR检测结果显示,和对照组相比,在LPS诱导后黑龙江林蛙各组织中的Fer-H基因转录水平均极显著上调(P<0.01)。综上所述,各组织Fer-H基因会通过上调其转录水平参与林蛙的抗氧化应激反应。本研究首次验证了LPS诱导下黑龙江林蛙各组织会出现氧化损伤,并为探究Fer-H基因与两栖类动物抗氧化反应之间的关系奠定了基础。In order to explore the mechanism of the Ferritin heavy chain subunit gene(Fer-H) of Rana amurensis participating in the body's antioxidant response under the induction of lipopolysaccharide(LPS),LPS-induced inflammation model of R.amurensis was established.1 m L LPS solution(1 mg/m L) was intraperitoneally injected into Heilongjiang wood frogs.After 6 hours,24 hours,48 hours,72 hours,and 120 hours of induction,samples were collected and HE staining was performed to observe the tissue lesions.The results showed that the liver,skin and muscle tissues of Heilongjiang forest frog were damaged under LPS-induced treatment,thus the LPS-induced forest frog inflammation model was proved feasible.The activities of antioxidant enzymes such as superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH) in tissues was measured,the changes of transcription level of Fer-H gene in different organs were detected by fluorescence quantitative PCR.Compared with the control group,the activities of SOD and CAT in the treatment group increased significantly(P0.01),while the activities of GSH decreased significantly(P0.01).The results of fluorescence quantitative PCR showed that the transcription level of Fer-H gene was significantly up-regulated after LPS induction(P0.01).In summary,LPS-induced oxidative damage occurred in R.amurensis,and the Fer-H gene would participate in the antioxidant stress response by up-regulating its transcription level.This study lays a foundation for exploring the relationship between Fer-H gene and amphibian antioxidant response.

关 键 词：黑龙江林蛙 脂多糖 氧化应激 铁蛋白H亚基基因 

分 类 号：S852.4[农业科学—基础兽医学]