藏猪(Sus scrofa)Ⅱ型肺泡上皮细胞低氧应答关键时间点筛选  

Screening of Critical Time Points for Hypoxic Response of Type Ⅱ Alveolar Epithelial Cells in Tibetan Pigs(Sus scrofa)

作　　者：席彬鹏 杨雅楠[1] 黄志强 牛瑞来 樊海 赵生国[1] XI Binpeng;YANG Yanan;HUANG Zhiqiang;NIU Ruilai;FAN Hai;ZHAO Shengguo(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou,730070)

机构地区：[1]甘肃农业大学动物科学技术学院,兰州730070

出　　处：《基因组学与应用生物学》2023年第7期736-743,共8页Genomics and Applied Biology

基　　金：国家自然科学基金(32060730);甘肃农业大学青年导师扶持基金(GAU-QDFC-2018-03);国家级大创计划项目(202010733015)共同资助。

摘　　要：为探究藏猪(Sus scrofa)Ⅱ型肺泡上皮细胞(typeⅡalveolar epithelial cells, ATⅡ)对低氧环境应答的关键时间点,本研究在常氧(21%O_(2))和低氧(2%O_(2))水平下培养藏猪和长白猪的ATⅡ,观察并分析不同时间点(0~144 h)ATⅡ的三维构象特征,内皮PAS结构域包含蛋白(endothelial PAS domain protein 1,EPAS1)和血红蛋白β(hemoglobin beta locus,HBB)基因的表达量,以及细胞增殖率的变化,寻找藏猪对低氧环境应答的关键时间点,并使用流式细胞仪分析其凋亡率。结果表明,成功分离到纯度达90%以上的藏猪ATⅡ,低氧培养的藏猪ATⅡ呈现明显的纤维化。0~36 h时,藏猪和长白猪EPAS1的表达量变化较为平缓,在48 h时表达量变化显著(P<0.05)。在低氧环境下,与长白猪相比,与长白猪相比,藏猪ATⅡ的EPAS1基因表达量更低,此变化趋势一直延续至96 h;HBB基因在藏猪和长白猪ATⅡ中的相对表达量随着时间的变化呈现先升高后降低的趋势,在低氧环境下,与长白猪相比,藏猪ATⅡ中更高;在低氧24~120 h时,藏猪ATⅡ在2%O_(2)培养下其细胞增殖率变化明显,且在培养120 h时是高于其他组的;48 h低氧组(藏猪ATⅡ2%O2培养,长白猪ATⅡ2%O_(2)培养)的ATⅡ晚期凋亡率均高于常氧组(藏猪ATⅡ21%O_(2)培养,长白猪ATⅡ21%O_(2)培养)的。结果表明,48 h是藏猪ATⅡ的低氧应答的关键时间点。To explore the critical time points of type Ⅱ alveolar epithelial cells(ATII)in response to hypoxia in Sus scrofa,in this study,AT Ⅱ of Tibetan pigs and Landrace pigs were cultured under normal oxygen(21%O_(2))and hypooxygen(2%O_(2))levels.The three-dimensional comformational characteristics of AT Ⅱ,the change in the gene expression levels of EPASI(endothelial PAS domain protein 1)and HBB(hemoglobin beta locus),and the proleferation rate of AT Ⅱ were observed and analyzed ATⅡ at different time points(0~144 h).The key change points of Tibetan pigs'response to hypoxic environment were investigated and the apoptosis rate was analyzed by flow cytometry.Results showed that ATⅡ of Tibetan pigs with purity of more than 90% was isolated successfully.ATⅡ of Tibetan pigs cultured with low oxygen showed obvious fibrosis.The expression level of EPASI in Tibetan pigs and Landrace pigs changed gently at 0~36 h.and significantly at 48 h(P<0.05).Compared to Landrace pigs,the expression level of EPASI gene in AT Ⅱ of Tibetan pigs was lower in hypoxia environment,and the change trend continued until 96 h.Compared to Landrace pigs,the rela-tive expression of HBB gene AT Ⅱ in the two groups firstly increased and then decreased with the change of time,and AT Ⅱ in Tibetan pigs was higher under hypoxia environment.At low oxygen levers of 24 h to 120 h,the cell proleferation rate of Tibetan pigs AT Ⅱ cultured with 2%O_(2),was significantly higher than that of other groups,and at 120 hours of cultivation,it was higher than other groups.The late apoptosis rate of AT Ⅱ in 48 h hypoxia group(Tibetan pigs ATⅡcultured under 2%O_(2),Landrace pigs AT Ⅱ cultured under 2%O_(2))was higher than that in normal oxygen group(Tibetan pigs AT Ⅱ cultured under 21%O_(2),Landrace pigs AT Ⅱ cultured under 21%O_(2)).The results showed that 48 h was the key time point for hypoxia response of Tibetan pigs AT Ⅱ.

关 键 词：藏猪 低氧适应 肺脏 Ⅱ型肺泡上皮细胞 

分 类 号：S828[农业科学—畜牧学]