辅助生殖技术对小鼠胎盘氨基酸转运通道载体蛋白表达的影响

Research on the protein expression of amino acid transport channel of mice placenta from assisted reproductive technology

目的探讨辅助生殖技术(ART)对胎盘氨基酸转运通道载体蛋白表达水平的影响。方法分别选取体外受精(IVF)18.5 d昆明仔鼠胎盘组织(IVF组)和自然妊娠18.5 d昆明仔鼠的胎盘组织(对照组),对两组出生体质量、胎盘相对重量及胎盘效率(出生体质量与胎盘相对重量比值)进行比较和统计分析,采用实时定量聚合酶链反应(RT-q PCR)及蛋白质印迹(Western blotting)方法检测两组胎盘组织中氨基酸转运体A系统(Snat1、Snat2、Snat4)及L系统(Lat1、Lat2)的m RNA和蛋白表达水平。结果 (1)IVF组新生仔鼠体质量[(1 609.14±128.23)mg]与对照组[(1 640.20±148.13)mg]间差异无统计学意义(P>0.05);IVF组小鼠胎盘相对重量[(192.86±28.87)mg]显著高于对照组[(126.20±21.50)mg](P=0.000);IVF组小鼠胎盘效率[(8.52±1.54)%]显著低于对照组胎盘效率[(13.38±2.73)%](P=0.000)。(2)与对照组相比,IVF组A系统氨基酸转运体Snat1、Snat2、Snat4 m RNA和其蛋白表达水平显著下调(m RNA:P=0.000、P=0.008、P=0.005;蛋白:P=0.008、P=0.006、P=0.000)。(3)与对照组相比,IVF组L系统氨基酸转运体Lat1 m RNA和蛋白表达水平差异无统计学意义(P>0.05);Lat2 m RNA和蛋白表达水平显著下调(P=0.011、P=0.000)。结论 ART导致小鼠足月胎盘氨基酸转运体Snat1、Snat2、Snat4及Lat2的表达水平显著下调,提示ART会影响妊娠末期胎盘氨基酸的转运能力。

Objective To study the effect of assisted reproductive technology（ART） on the protein expression of amino acid transport channel in placenta. Methods We chose the placenta tissue of mice by in vitro fertilization（IVF） in 18.5 d（IVF group） and by natural pregnancy in 18.5 d（control group） to compare the birth weight, placental relative weight and ratio of the fetal/placental weight. We then detected the m RNA and protein expressions of placental amino acid transporter（Snat1, Snat2, Snat4, Lat1 and Lat2） by realtime quantitative polymerase chain reaction（RT-q PCR） and Western blotting. Results 1） There were no statistical differences in new birth weight between IVF group [（1 609.14±128.23） mg] and control group [（1 640.20±148.13） mg]（P〈0.05）. Compared with control group [（126.20±21.50） mg,（13.38±2.73）%], the placenta relative weight of mice was significantly higher in IVF group [（192.86±28.87） mg, P=0.000] and ratio of the fetal/placental weight was significantly lower in IVF group [（8.52±1.54）%]（P=0.000）. 2） Compared with control group, the m RNA and protein expression of A system amino acid transporter Snat1, Snat2 and Snat4 in placenta were significantly down-regulated in IVF group（m RNA： P=0.000, P=0.008, P=0.005; protein： P=0.008, P=0.006, P=0.000）. 3） There were no statistical differences of the m RNA and protein expression of L system amino acid transporter Lat1 between these two groups（P〈0.05）, but the m RNA and protein expression of amino acid transporter Lat2 in placenta was significantly down-regulated in IVF group（P=0.011, P=0.000）. Conclusion ART leads to the expressions of amino acid transporter Snat1, Snat2, Snat4 and Lat2 significantly down-regulated in placenta, which suggests that ART may affect the function of placenta amino acid transporter during the late pregnancy.