塔里木马鹿抗氧化基因 PRDX 1的功能初探  

Preliminary Study on the Function of PRDX 1 Antioxidant Gene in Tarim Red Deer

作　　者：布威海丽且姆·阿巴拜科日 艾合麦提尼亚孜·艾合麦提江 乃比江·麦图荪 单文娟 ABABAIKERI Buweihailiqiemu;AIHEMAITIJIANG Aihemaitiniyazi;MOHAMMADTURSUN Nabijan;SHAN Wenjuan(Xinjiang Hetian College,Hotan 848000,China;Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830017,China;Xinjiang Key Laboratory of Hotan Characteristic Chinese Traditional Medicine Research,Hotan 848000,China;College of Life Sciences,Wuhan University,Wuhan 430072,China)

机构地区：[1]新疆和田学院,和田848000 [2]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,乌鲁木齐830017 [3]新疆和田特色中医药研究重点实验室,和田848000 [4]武汉大学生命科学学院,武汉430072

出　　处：《畜牧兽医学报》2025年第3期1216-1230,共15页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：新疆维吾尔自治区自然科学基金项目(2022D01B40)。

摘　　要：旨在探讨塔里木马鹿抗氧化功能基因过氧化物酶1(PRDX 1)在氧化应激中对细胞的保护作用。本研究利用塔里木马鹿PRDX 1基因(CeyPRDX 1)过表达载体(pLJM1-CeyPRDX 1-EGFP)感染人永生化角质形成细胞HaCaT后,通过MTT法建立HaCaT细胞在高温、盐(NaCl)和过氧化氢(H 2O 2)等胁迫条件下的损伤模型。在这3种氧化应激胁迫条件下检测过表达CeyPRDX 1基因对细胞的凋亡率、线粒体膜电位变化(JC-1)、活性氧(ROS)含量、丙二醛(MDA)含量以及SOD1、PTEN和Caspase-3等基因mRNA相对表达量的影响。结果表明,将200 mmol·L^(-1)的NaCl干预细胞24 h、0.50 mmol·L^(-1)的H 2O 2处理细胞12 h和42℃培养细胞4 h选为细胞氧化损伤的最佳胁迫条件。在此胁迫条件下,过表达CeyPRDX 1基因能够增加细胞存活率,并减少由NaCl、H 2O 2和高温等胁迫条件诱导的细胞凋亡率、线粒体膜电位的损伤、ROS以及MDA含量,从而有效降低氧化应激诱导的细胞损伤。qRT-PCR结果表明,在3种胁迫条件下,与未感染和感染空载慢病毒的HaCaT细胞相比,感染CeyPRDX 1过表达慢病毒的HaCaT细胞中SOD 1的mRNA表达量显著升高(P<0.001),而PTEN和凋亡相关基因Caspase-3的mRNA表达量显著下降(P<0.01)。本研究结果表明,CeyPRDX 1基因可能在塔里木马鹿适应极端干旱环境和耐受高盐饮食的过程中起到保护细胞免受氧化应激损伤的作用。其作用可能与SOD1、PTEN和Caspase-3等基因的表达调控有关。This study was conducted to investigate the protective effect of the antioxidant gene peroxiredoxin I(PRDX 1)of Tarim red deer in cells under oxidative stress.The Tarim red deer PRDX1(CeyPRDX 1)overexpression vector(pLJM1-CeyPRDX 1-EGFP)was used to infect human immortalized keratinocyte HaCaT cells,and MTT method was used to establish the injury model of HaCaT cells under high temperature,salt(NaCl)and hydrogen peroxide(H_(2)O_(2))stress conditions.Under this stress conditions,the effects of overexpression of CeyPRDX 1 on cell apoptosis rate,mitochondrial membrane potential changes(JC-1),reactive oxygen species(ROS)content,malondialdehyde(MDA)content,and relative expression levels of SOD1,PTEN,and Caspase-3 genes were examined.The results showed that the 24 h of intervention with 200 mmol·L^(-1) NaCl,12 h of treatment with 0.50 mmol·L^(-1) H_(2)O_(2),and 4 h of cultivation at 42℃ were selected as the optimal stress conditions for cellular oxidative damage.Under stress conditions,overexpression of CeyPRDX 1 gene could increase cell survival rate,and reduce apoptosis rate,mitochondrial membrane potential damage,ROS and MDA content induced by stress conditions such as NaCl,H_(2)O_(2) and high temperature,effectively reducing oxidative stress-induced cell damage.The qRT-PCR results showed that under 3 stress conditions,compared to HaCaT cells that were not infected and HaCaT cells infected with empty lentivirus,HaCaT cells infected with CeyPRDX 1 overexpressing lentivirus showed significantly increased mRNA expression level of SOD1(P<0.001),while significantly decreased mRNA expression levels of PTEN and apoptosis related gene Caspase-3(P<0.01).The results indicate that CeyPRDX 1 gene may play an important role in protecting cells from oxidative stress damage in the adaptation of Tarim red deer to extreme drought environments and tolerance to high salt diets.Its role may be related to the expression regulation of genes such as SOD1,PTEN and Caspase-3.

关 键 词：塔里木马鹿 PRDX 1基因 HACAT细胞 CeyPRDX 1过表达 氧化应激 

分 类 号：S825.2[农业科学—畜牧学]