宫内发育迟缓猪胎盘SLC7A2基因过表达对滋养层细胞增殖和迁移的影响  

Effects of SLC7A2 gene overexpression on trophoblast cell proliferation and migration in the placenta of pigs with intrauterine growth retardation

作　　者：胡光玲 伍治敏 张才在 杨振清 张依裕 敖政 HU Guang-ling;WU Zhi-min;ZHANG Cai-zai;YANG Zhen-qing;ZHANG Yi-yu;AO Zheng(College of Animal Science,Guizhou University/Key Laboratory of Animal Genetics,Breeding and Reproduction in thePlateau Mountainous Region,Ministry of Education/Guizhou Provincial Key Laboratory of Animal Genetics,Breedingand Reproduction,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州大学动物科学学院/高原山地动物遗传育种与繁殖教育部重点实验室/贵州省动物遗传育种与繁殖重点实验室,贵州贵阳550025

出　　处：《南方农业学报》2024年第6期1778-1786,共9页Journal of Southern Agriculture

基　　金：贵州省科技计划项目(黔科合基础-ZK〔2021〕一般166);贵州省科技人才项目(黔科合平台人才〔2021〕5630号);贵州省普通高等学校青年科技人才成长项目(黔教合KY字〔2021〕081号);贵州省生猪产业发展项目(黔财农〔2021〕157号)。

摘　　要：【目的】探究溶质载体家族7成员2基因(SCL7A2)在正常初生重(NBW)和宫内发育迟缓(IUGR)猪胎盘中的表达差异,明确SCL7A2基因过表达对猪滋养层细胞(pTr2)增殖和迁移的影响,为揭示猪IUGR的发生发展机制提供参考依据。【方法】采用实时荧光定量PCR检测NBW和IUGR胎盘中SLC7A2基因相对表达量,通过免疫组织化学试验分析NBW和IUGR胎盘组织中SLC7A2蛋白的定位分布及表达情况,利用CCK-8试验检测SLC7A2基因过表达对pTr2细胞增殖能力的影响,并以细胞划痕试验检测SLC7A2基因过表达对pTr2细胞迁移能力的影响。【结果】IUGR胎盘中的SLC7A2基因相对表达量极显著高于NBW胎盘(P<0.01,下同);SLC7A2蛋白在仔猪胎盘的滋养层细胞和血管内皮细胞中均有表达,且IUGR胎盘中的SLC7A2蛋白表达水平显著高于NBW胎盘(P<0.05,下同)。以构建的SLC7A2基因过表达载体(Vector-SLC7A2+)转染pTr2细胞,细胞中的SLC7A2基因相对表达量较pEGFP-C1阴性对照载体(Vector-NC)转染组极显著上调,但pEGFP-C1的相对表达量与Vector-NC转染组无显著差异(P>0.05)。以Vector-SLC7A2+和Vector-NC分别转染pTr2细胞后,Vector-SLC7A2+转染组的pTr2细胞增殖效果在转染后12、24、48和72 h显著或极显著低于Vector-NC转染组,pTr2细胞迁移率则表现为Vector-SLC7A2+转染组极显著低于Vector-NC转染组,表明SLC7A2基因过表达能有效抑制pTr2细胞的增殖和迁移能力。【结论】SLC7A2基因过表达会抑制猪胎盘滋养层细胞的增殖与迁移,影响胎盘的形态发育和母—胎间的营养转运效率,进而诱发猪IUGR的发生发展。【Objective】This study aimed to investigate the expression differences of solute carrier family 7 member 2 gene(SCL7A2)in the placentas of normal birth weight(NBW)piglets and intrauterine growth retardation(IUGR)pig lets,and determine the effect of SCL7A2 gene overexpression on the proliferation and migration of trophoblast cells(pTr2)to provide a reference for revealing the occurrence and development mechanism of IUGR in pigs.【Method】Realtime fluorescence quantitative PCR(RT-qPCR)was used to detect the relative expression of SLC7A2 gene in NBW and IUGR placentas.The localization,distribution and expression level of SLC7A2 protein in placental tissues of NBW and IUGR were analyzed by immunohistochemistry.The effects of SLC7A2 gene overexpression on the proliferation were de tected by CCK-8 assay,effects of SLC7A2 gene overexpression on migration ability of pTr2 cells were detected by cell scratch assay.【Result】The relative expression of SLC7A2 gene in IUGR placentas was extremely significantly higher(P<0.01,the same bellow)than that in NBW placentas.SLC7A2 protein was expressed in both trophoblast cells and vascular endothelial cells in pig placentas,with a significantly higher expression level in IUGR placentas compared to NBW pla centas(P<0.05,the same bellow).The constructed SLC7A2 gene overexpression vector(Vector-SLC7A2+)was trans fected into pTr2 cells,resulting in an extremely significant up-regulation of the relative expression of SLC7A2 gene in the cells compared to the pEGFP-C1 negative control vector(Vector-NC)transfected group.However,there was no signifi cant difference in the relative expression of pEGFP-C1 between the two groups(P>0.05).After transfection of pTr2 cells with Vector-SLC7A2+and Vector-NC respectively,the proliferation of pTr2 cells in the Vector-SLC7A2+transfected group was significantly or extremely significantly lower than that in the Vector-NC transfected group at 12,24,48,and 72 h post-transfection.Similarly,the migration rate of pTr2 cells in the Vector-SLC7A2+transfec

关 键 词：猪 SLC7A2基因 胎盘 宫内发育迟缓(IUGR) 营养转运 

分 类 号：S828.3[农业科学—畜牧学]