空气诱导小鼠子宫内膜蜕膜化
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摘要
目的探讨空气对小鼠子宫内膜蜕膜化的诱导作用,建立非手术法小鼠子宫内膜蜕膜化诱导技术。方法利用小鼠非手术法胚胎移植器,通过子宫颈向30只假孕3.5 d小鼠单侧子宫角移植2μl空气,诱导子宫内膜蜕膜化,观察蜕膜化子宫形态并测量其直径和重量,HE染色观察细胞形态,RT-PCR检测蜕膜化相关标记基因表达。结果移植空气诱导25只假孕小鼠出现不同程度的蜕膜化现象。与对照侧子宫角相比,空气移植侧子宫角明显肿大、增粗、增重,子宫内膜毛细血管通透性增加,蜕膜化标记基因环氧化物水解酶-2(COX-2)、泌乳素(Prl)3c1、Prl8a2等高表达,且细胞出现多核现象。结论利用非手术法子宫内移植空气可以高效诱导小鼠子宫内膜蜕膜化。
Objective To establish a non-surgical induced decidualization technique in mice,and to explore the air-induced decidua in this study. Methods Clean air( 2 μl) was transferred into unilateral uterine horn of 30 pseudopregnant 3. 5 days mice by a non-surgical transfer method. The morphology, weight, diameter, cellular differentiation,and decidua-related marker genes in air-induced decidua were detected. Results After air transfer four days,twenty-five pseudopregnant mice displayed different degree of decidualization phenomenon. Compared to the control side uterine horn,air-transferred uterine horns appeared significantly dilation and overweight with increased diameter. The permeability of uterine endometria was extensively spread along the whole horns. Three decidualization-realated marker genes [cyclooxygenase-2( COX-2),prolactin( Prl) 3 c1 and Prl8 a2] had a higher expression level. In addition,polykaryocytes was observed in air-induced decidua. Conclusion Mouse uterine endometrial decidualization can be effectively induced by a non-surgical air transfer method.
Objective To establish a non-surgical induced decidualization technique in mice,and to explore the air-induced decidua in this study. Methods Clean air( 2 μl) was transferred into unilateral uterine horn of 30 pseudopregnant 3. 5 days mice by a non-surgical transfer method. The morphology, weight, diameter, cellular differentiation,and decidua-related marker genes in air-induced decidua were detected. Results After air transfer four days,twenty-five pseudopregnant mice displayed different degree of decidualization phenomenon. Compared to the control side uterine horn,air-transferred uterine horns appeared significantly dilation and overweight with increased diameter. The permeability of uterine endometria was extensively spread along the whole horns. Three decidualization-realated marker genes [cyclooxygenase-2( COX-2),prolactin( Prl) 3 c1 and Prl8 a2] had a higher expression level. In addition,polykaryocytes was observed in air-induced decidua. Conclusion Mouse uterine endometrial decidualization can be effectively induced by a non-surgical air transfer method.
引文
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[6]Zhan S,Lin HY,Kong SB,et al.Physiological and molecular determinants of embryo implantation[J].Mol Aspects Med,2013,34(5):939-980.
[7]Das SK,Lim H,Paria BC,et al.Cyclin D3 in the mouse uterus is accociated with decidulization process during early pregnancy[J].J Mol Endocrinol,1999,22(1):91-101.
[8]Duan XCh,Li Y,Di KQ,et al.A nonsurgical embro transfer transfer technique in mice[J].Chinese Journal of Biotechnology,2016,32(4):440-446.(in Chinese)段新崇,李阳,邸科前,等.一种非手术性小鼠胚胎移植技术[J].生物工程学报,2016,32(4):440-446.
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[10]Krampl E,Zegermacher G,Eichler C,et al.Air in the uterine cavity after embryo transfer[J].Fertil Steril,1995,63(2):366-370.
[11]Tiras B,Korucuoglu U,Polat M,et al.Effect of air bubble localization after transfer on embryo transfer outcomes[J].Eur J Obstet Gynecol Reprod Biol,2012,164(1):52-54.
[12]Tan J,Raja S,Davis MK,et al.Evidence for coordinated interaction of cyclin D3 with P21 and cdk6 in directing the development of uterine stromal cell decidulazation and polyploidy during implantation[J].Mech Dev,2002,111(1-2):99-113.
[13]Hao J,Gao YM,Liu K,et al.Immunolocalization of TGFβ1and its receptor in mouse endometium during the process of implantion[J].Acta Anatomica Sinica,2001,32(1):64-67.(in Chinese)郝晶,高英茂,刘凯,等.转化生长因子β1及其受体在着床期小鼠子宫内膜中的免疫组织化学研究[J].解剖学报,2001,32(1):64-67.
[14]Zhang X,Fu LJ,Liu XQ,et al.Nm23 regulates decidualization through the PI3K-Akt-m TOR signaling pathways in mice and humans[J].Hum Reprod,2016,31(10)1-13.
[15]Zhang D,Chang X,Bai J,et al.The study of cyclooxygenase 2 in human decidua of preeclampsia[J].Biol Reprod,2016,95(3):56.
[2]Yu YSh,Tian Zh,Luo XT,et al.Study on the induced decidualization of mouse endometrium[J].Journal of Anhui Agricultural Sciences,2008,36(20):8615-8616,8644.(in Chinese)于永生,田振,罗晓彤,等.小鼠子宫内膜诱导蜕膜化的研究[J].安徽农业科学,2008,36(20):8615-8616,8644.
[3]Herington JL,Bany BM.Do molecular signals from the conceptus influence endometrium decidualization in rodents[J]?J Exp Zool B Mol Dev Evol,2011,312(8):797-816.
[4]Orsini MW.Attempted decidualization in the hamster and rat with pyrathiazine[J].J Reprod Fertil,1963,5(3):323-330.
[5]Orsini MW,IDonovan BT.Implantation and induced decidualization of the uterus in the guinea pig as indicated by pontamine blue[J].Biol Reprod,1971,5(3):270-281.
[6]Zhan S,Lin HY,Kong SB,et al.Physiological and molecular determinants of embryo implantation[J].Mol Aspects Med,2013,34(5):939-980.
[7]Das SK,Lim H,Paria BC,et al.Cyclin D3 in the mouse uterus is accociated with decidulization process during early pregnancy[J].J Mol Endocrinol,1999,22(1):91-101.
[8]Duan XCh,Li Y,Di KQ,et al.A nonsurgical embro transfer transfer technique in mice[J].Chinese Journal of Biotechnology,2016,32(4):440-446.(in Chinese)段新崇,李阳,邸科前,等.一种非手术性小鼠胚胎移植技术[J].生物工程学报,2016,32(4):440-446.
[9]Cui L,Zhang Z,Sun F,et al.Transcervical embryo transfer in mice[J].J Am Assoc Lab Anim Sci,2014,53(3):228-231.
[10]Krampl E,Zegermacher G,Eichler C,et al.Air in the uterine cavity after embryo transfer[J].Fertil Steril,1995,63(2):366-370.
[11]Tiras B,Korucuoglu U,Polat M,et al.Effect of air bubble localization after transfer on embryo transfer outcomes[J].Eur J Obstet Gynecol Reprod Biol,2012,164(1):52-54.
[12]Tan J,Raja S,Davis MK,et al.Evidence for coordinated interaction of cyclin D3 with P21 and cdk6 in directing the development of uterine stromal cell decidulazation and polyploidy during implantation[J].Mech Dev,2002,111(1-2):99-113.
[13]Hao J,Gao YM,Liu K,et al.Immunolocalization of TGFβ1and its receptor in mouse endometium during the process of implantion[J].Acta Anatomica Sinica,2001,32(1):64-67.(in Chinese)郝晶,高英茂,刘凯,等.转化生长因子β1及其受体在着床期小鼠子宫内膜中的免疫组织化学研究[J].解剖学报,2001,32(1):64-67.
[14]Zhang X,Fu LJ,Liu XQ,et al.Nm23 regulates decidualization through the PI3K-Akt-m TOR signaling pathways in mice and humans[J].Hum Reprod,2016,31(10)1-13.
[15]Zhang D,Chang X,Bai J,et al.The study of cyclooxygenase 2 in human decidua of preeclampsia[J].Biol Reprod,2016,95(3):56.