猪胎盘营养转运相关基因在不同妊娠期的表达

Expression of Nutrition Transport-related Genes in Porcine Placenta at Different Gestation Periods

【目的】探究不同妊娠时期猪胎盘的氨基酸、葡萄糖、脂肪酸转运体的表达模式。【方法】选择15头遗传背景、产仔数接近的杜洛克2~4胎经产健康母猪平均分为3组,所有母猪发情后使用相同公猪精液进行人工授精,在妊娠第40天(D40)、65天(D65)和95天(D95)通过麻醉分别取出每组母猪子宫,快速打开子宫分离出每个胎儿的胎盘组织,提取胎盘组织总RNA并反转录合成cDNA,利用合成的引物进行普通PCR扩增,用2.0%琼脂糖凝胶检测扩增产物。采用实时荧光定量PCR检测并比较3个时期胎盘中氨基酸、葡萄糖、脂肪酸转运体相关基因mRNA相对表达水平。【结果】PCR检测结果显示,氨基酸转运体相关基因(SLC7A1、SLC7A2、SLC7A3、SLC7A4、SLC7A10、SLC1A3、SLC1A5、SLC38A10、SLC36A1)、葡萄糖转运体相关基因(SLC2A1、SLC2A2、SLC2A3、SLC2A10、SLC2A12、SLC2A13)及脂肪酸转运体相关基因(FATP1、FATP2、FATP3、FATP4、FABP3、FABP5、FABP7、CD36)的片段长度均与预期相符。实时荧光定量PCR结果显示,在氨基酸转运体中,D65胎盘中SLC7A4、SLC7A10、SLC38A10基因表达水平显著高于D40胎盘(P<0.05),而SLC7A2基因表达水平显著低于D40胎盘(P<0.05),且D65胎盘的SLC1A3和SLC7A4基因表达水平均显著低于D95胎盘(P<0.05);在葡萄糖转运体中,D65和D95胎盘的SLC2A3和SLC2A13基因表达水平显著高于D40胎盘(P<0.05),D95胎盘的SLC2A1、SLC2A2和SLC2A12基因表达水平显著低于D65胎盘(P<0.05);在脂肪酸转运体中,D65胎盘的FATP2、FATP4、FABP3、FABP5、FABP7和CD36基因表达水平显著高于D40胎盘(P<0.05),而FATP1、FATP4和CD36基因表达水平显著低于D95胎盘(P<0.05)。【结论】在猪妊娠过程中,胎盘中SLC7A10、SLC38A10、SLC7A4、SLC2A3、FATP1、FATP4、FABP5、CD36等基因可能是影响胎儿生长发育的重要营养转运基因。

【Objective】The aim of this study was to explore the expression patterns of amino acids,glucose and fatty acid transporters in porcine placenta at different gestation periods.【Method】15 Duroc sows with similar genetic background and litter size were randomly divided into 3 groups.After estrus,all sows were artificially inseminated with semen collected from the same Duroc boar.The uteruses of sows in each group were taken out by anesthesia on day 40(D40),65(D65)and 95(D95)of gestation.The uterus was quickly opened to separate the placental tissue of each fetus and the total RNA of placental tissue was extracted and synthesized cDNA.The synthesized primers were used for general PCR amplification and 2.0%agarose gel was used to detect the amplification products.Real-time quantitative PCR was used to detect and compare the relative mRNA expression levels of amino acid,glucose,fatty acid transporters associated genes in placenta at three pregnancy stages.【Result】PCR results showed that the length of amino acid transporter-related genes(SLC7 A1,SLC7 A2,SLC7 A3,SLC7 A4,SLC7 A10,SLC1 A3,SLC1 A5,SLC38 A10,SLC36 A1),glucose transporter related genes(SLC2 A1,SLC2 A2,SLC2 A3,SLC2 A10,SLC2 A12,SLC2 A13),and fatty acid transporter-related genes(FATP1,FATP2,FATP3,FATP4,FABP3,FABP7,CD36)were consistent with the expected.Real-time quantitative PCR results showed that in amino acid transporters,the expression of SLC7A4,SLC7A10 and SLC38A10 genes in D65 placenta were significantly higher than those in D40 placenta(P<0.05),while SLC7A2 gene was significantly lower than that in D40 placenta(P<0.05),and the expression SLC1A3 and SLC7A4 genes in D65 placenta were significantly lower than those in D95 placenta(P<0.05).In glucose transporters,the expression of SLC2A3 and SLC2A13 genes in D65 and D95 placenta were significantly higher than those in D40 placenta,while the expression of SLC2A1,SLC2A2 and SLC2A12 genes in D95 placenta were significantly lower than those in D65 placenta(P<0.05).In fatty acid transporters,the expression of FATP2,FATP4,FABP3,FABP5,FABP7 and CD36 genes in D65 placenta were significantly higher than those in D40 placenta(P<0.05),while the expression of FATP1,FATP4 and CD36 genes were significantly lower than those in D95 placenta(P<0.05).【Conclusion】The placental SLC7A10,SLC38A10,SLC7A4,SLC2A3,FATP1,FATP4,FABP7 and CD36 might be nutrition transport-related genes affecting fetal growth and development in porcine pregnancy.