金属蛋白硫在体外受精-胚胎移植胎盘及胚胎组织中表达及意义
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摘要
目的研究辅助生殖中体外受精-胚胎移植(IVF-ET)技术本身对早期胎盘绒毛和胚胎组织中金属蛋白硫及其基因表达影响。方法收集辅助生殖IVF-ET来源,双胚胎移植妊娠45～60天,经超声引导下减胎获得胎盘绒毛和胚胎组织各8例,作为研究组,对照组采用自然妊娠双胎45～60天人工流产中获得的胎盘绒毛和胚胎组织各10例。从采用免疫组织化学方法和实时荧光定量RT-PCR方法检测组织中金属蛋白硫蛋白和mRNA的表达及分布差异。结果表达部位:金属蛋白硫表达于胎盘绒毛细胞滋养层细胞和合体滋养层细胞的细胞膜和细胞质,广泛表达于胚胎各个胚层。表达特点:IVF-ET来源胎盘绒毛组织中金属蛋白硫表达的吸光度值显著低于自然妊娠组,差异有统计学意义。金属蛋白硫在IVF-ET和自然妊娠胚胎组织中表达比较,差异无统计学意义。基因表达:金属蛋白硫mRNA在IVF-ET胎盘绒毛组织中低表达,而在胚胎组织中表达比较,差异无统计学意义,与免疫组织化学结果相一致。结论金属蛋白硫蛋白和基因在IVF-ET胎盘绒毛组织中表达低于自然妊娠组织,而在胚胎组织中表达比较,差异无统计学意义,显示IVF-ET技术本身更加倾向于影响胎盘发育和功能,金属蛋白硫可能在IVF-ET相关疾病发病机制中发挥作用。
Objective To study the effect of IVF-ET on the expression of metallothionein and its gene in placenta villus and embryonic tissues. Methods In vitro fertilization-embryo transfer patients were collected and transplanted 45-60 days after pregnancy by double embryo transfer. Ultrasound-guided embryo reduction was performed to obtain 8 cases of placental villus and embryonic tissue. As the study group, the control group used natural pregnancy twins 45-60 days abortion. Ten cases of placental villus and embryonic tissue were obtained. The expression and distribution of metallothionein protein and mRNA in tissues were detected by immunohistochemistry and real-time fluorescence quantitative RT-PCR. Results Metallothionein was expressed in the cell membrane and cytoplasm of placental villous trophoblast cells and syncytiotrophoblasts, and metallothionein was widely expressed in embryonic germ layers. The optical density of metallothionein expression in IVF-ET-derived placenta villi was significantly lower than that in natural pregnancy group. The difference was statistically significant. There was no significant difference in the expression of metallothionein between IVF-ET and natural pregnancy embryos. Metallothionein mRNA was lowly expressed in IVF-ET placenta villi, but there was no significant difference in embryonic tissue expression, consistent with immunohistochemical results. Conclusion The expression of metallothionein protein and gene in IVF-ET placenta villi was lower than that in natural pregnancy tissues, but there was no significant difference in embryonic tissues. It was shown that IVF-ET itself was more likely to affect placental development and function. Metallothionein may play a role in the pathogenesis of IVF-ET related diseases.
Objective To study the effect of IVF-ET on the expression of metallothionein and its gene in placenta villus and embryonic tissues. Methods In vitro fertilization-embryo transfer patients were collected and transplanted 45-60 days after pregnancy by double embryo transfer. Ultrasound-guided embryo reduction was performed to obtain 8 cases of placental villus and embryonic tissue. As the study group, the control group used natural pregnancy twins 45-60 days abortion. Ten cases of placental villus and embryonic tissue were obtained. The expression and distribution of metallothionein protein and mRNA in tissues were detected by immunohistochemistry and real-time fluorescence quantitative RT-PCR. Results Metallothionein was expressed in the cell membrane and cytoplasm of placental villous trophoblast cells and syncytiotrophoblasts, and metallothionein was widely expressed in embryonic germ layers. The optical density of metallothionein expression in IVF-ET-derived placenta villi was significantly lower than that in natural pregnancy group. The difference was statistically significant. There was no significant difference in the expression of metallothionein between IVF-ET and natural pregnancy embryos. Metallothionein mRNA was lowly expressed in IVF-ET placenta villi, but there was no significant difference in embryonic tissue expression, consistent with immunohistochemical results. Conclusion The expression of metallothionein protein and gene in IVF-ET placenta villi was lower than that in natural pregnancy tissues, but there was no significant difference in embryonic tissues. It was shown that IVF-ET itself was more likely to affect placental development and function. Metallothionein may play a role in the pathogenesis of IVF-ET related diseases.
引文
1 de Ziegler D,Pirtea P,Poulain M,et al.Time to think about neonatal outcome in assisted reproductive technology[J].Fertil Steril,2018,109(5):789-7901
2 Sekovani■,Jurasovi■ gene polymorphism and trace elements in mother-newborn pairs in the Croatian population[J].J Trace Elem Med Biol,2018,45(5):163-170
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4 Purewal S,Chapman SCE,van den Akker OBA.Depression and state anxiety scores during assisted reproductive treatment are associated with outcome:a meta-analysis[J].Reprod Biomed Online,2018,36(6):646-657
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6 Horie M,Shimizu K,Tabei Y.Validation of metallothionein,interleukin-8,and heme oxygenase-1 as markers for evaluation of cytotoxicity caused by metal oxide nanoparticles[J].Toxicol Mech Methods,2018,8(1):1-25
7 Mairinger FD,Schmeller J,Borchert S,et al.Immunohistochemically detectable metallothionein expression in malignant pleural mesotheliomas is strongly associated with early failure to platin-based chemotherapy[J].Oncotarget,2018,9(32):22254-22268
8 Abril SIM,Costa PG,Bianchini A.Metal accumulation and expression of genes encoding for metallothionein and copper transporters in a chronically exposed wild population of the fish Hyphessobrycon luetkenii[J].Comp Biochem Physiol C Toxicol Pharmacol,2018,211(8):25-31
9 Kalsotra T,Khullar S,Agnihotri R,et al.Metal induction of two metallothionein genes in the ectomycorrhizal fungus Suillus himalayensis and their role in metal tolerance[J].Microbiology,2018,164(6):868-876
10 Fabrin TMC,Diamante NA,Mota TFM,et al.Performance of biomarkers metallothionein and ethoxyresorufin O-deethylase in aquatic environments:a meta-analytic approach[J].Chemosphere,2018,205:339-349
11 Baldwin HJ,Patterson JA,Nippita TA,et al.Antecedents of abnormally invasive placenta in primiparous women:risk associated with gynecologic procedures[J].Obstet Gynecol,2018,131(2):227-233
12 Wang X,Wu SP,DeMayo FJ.Hormone dependent uterine epithelial-stromal communication for pregnancy support[J].Placenta,2017,60(11):S20-S26
13 Duranthon V,Chavatte-Palmer P.Long term effects of ART:what do animals tell us?[J].Mol Reprod Dev,2018,85(4):348-368
14 Li C,Zhang Y,Tang L,et al.Expression of factors involved in the regulation of angiogenesis in the full-term human placenta:effects of in vitro fertilization[J].Reprod Biol,2016,16(2):104-112
15 Shi XM,Wang Y,Zhang Y,et al.Effect of primary elective cesarean delivery on placenta accreta:a case-control study[J].Chin Med J(Engl),2018,131(6):672-676
2 Sekovani■,Jurasovi■ gene polymorphism and trace elements in mother-newborn pairs in the Croatian population[J].J Trace Elem Med Biol,2018,45(5):163-170
3 de Ziegler D,Andersen CY,Stanczyk FZ,et al.Endocrine mechanisms and assay issues in premature progesterone elevation in assisted reproductive technology[J].Fertil Steril,2018,109(4):571-576
4 Purewal S,Chapman SCE,van den Akker OBA.Depression and state anxiety scores during assisted reproductive treatment are associated with outcome:a meta-analysis[J].Reprod Biomed Online,2018,36(6):646-657
5 Henningsen AA,Bergh C,Skjaerven R,et al.Trends over time in congenital malformations in live-born children conceived after assisted reproductive technology[J].Acta Obstet Gynecol Scand,2018,97(7):816-823
6 Horie M,Shimizu K,Tabei Y.Validation of metallothionein,interleukin-8,and heme oxygenase-1 as markers for evaluation of cytotoxicity caused by metal oxide nanoparticles[J].Toxicol Mech Methods,2018,8(1):1-25
7 Mairinger FD,Schmeller J,Borchert S,et al.Immunohistochemically detectable metallothionein expression in malignant pleural mesotheliomas is strongly associated with early failure to platin-based chemotherapy[J].Oncotarget,2018,9(32):22254-22268
8 Abril SIM,Costa PG,Bianchini A.Metal accumulation and expression of genes encoding for metallothionein and copper transporters in a chronically exposed wild population of the fish Hyphessobrycon luetkenii[J].Comp Biochem Physiol C Toxicol Pharmacol,2018,211(8):25-31
9 Kalsotra T,Khullar S,Agnihotri R,et al.Metal induction of two metallothionein genes in the ectomycorrhizal fungus Suillus himalayensis and their role in metal tolerance[J].Microbiology,2018,164(6):868-876
10 Fabrin TMC,Diamante NA,Mota TFM,et al.Performance of biomarkers metallothionein and ethoxyresorufin O-deethylase in aquatic environments:a meta-analytic approach[J].Chemosphere,2018,205:339-349
11 Baldwin HJ,Patterson JA,Nippita TA,et al.Antecedents of abnormally invasive placenta in primiparous women:risk associated with gynecologic procedures[J].Obstet Gynecol,2018,131(2):227-233
12 Wang X,Wu SP,DeMayo FJ.Hormone dependent uterine epithelial-stromal communication for pregnancy support[J].Placenta,2017,60(11):S20-S26
13 Duranthon V,Chavatte-Palmer P.Long term effects of ART:what do animals tell us?[J].Mol Reprod Dev,2018,85(4):348-368
14 Li C,Zhang Y,Tang L,et al.Expression of factors involved in the regulation of angiogenesis in the full-term human placenta:effects of in vitro fertilization[J].Reprod Biol,2016,16(2):104-112
15 Shi XM,Wang Y,Zhang Y,et al.Effect of primary elective cesarean delivery on placenta accreta:a case-control study[J].Chin Med J(Engl),2018,131(6):672-676