异体子宫移植的动物实验研究

发布时间：2018-05-19 20:50

本文选题：子宫移植 + 动物实验　；参考：《中国人民解放军医学院》2017年博士论文

【摘要】：研究背景：完全性子宫因素不孕(abso1ute uterine factor-inferti1ity AUFI)是女性不孕因素的一个主要类型,一直被认为无法医治,主要是因为这类女性先天性无子宫比如MRKH (Mayer-Rokitansky-Kuster-Hauser)综合征患者或早期因为宫颈癌、产科出血被迫切除子宫和因为严重的子宫内粘连导致没有生育功能的育龄期女性,子宫移植(uterus transp1antation-UTx)的目的就是为先天或后天性因素导致子宫缺如或子宫畸形的育龄期女性移植一个有功能的子宫并生出一个孩子。人体子宫移植技术如果想要达到更安全的妊娠及活产仍需要大量的动物实验研究来提供技术支持,在国内,异体子宫移植的大动物实验研究一直处于探索阶段,鲜有相关研究报道;另外,在临床领域,育龄期子宫因素不孕女性对UTx的态度与意愿是不知的。研究目的:1.探讨小型猪异体子宫移植活体捐赠技术的可行性和安全性,观察子宫移植后动物的长期存活及生殖功能恢复;2.探讨称猴异体子宫移植活体捐赠的手术技巧,评估猕猴子宫肌层组织耐受冷缺血损伤的时限;3.调查MRKH综合征患者对子宫移植的态度和意愿,为下一步人体子宫移植提供临床数据和志愿者。方法:1.选用10头雌性、月经规律的小型猪,根据一母同源(同胞生)和体重匹配5头作为供体,5头作为受体(n=5),从活体供体盆腔获取子宫及主要血管,使用4℃C器官保存液对离体器官冷保存、灌注,之后将供体子宫原位移植到受体盆腔,移植的髂内动脉与受体髂外动脉端-侧吻合;2.术后5例受体均给予三联免疫抑制剂维持治疗,术后1周和1个月行多普勒超声观察移植子宫的动脉血流信号及盆腔情况,术后2个月全麻下行二次开腹观察移植子宫及盆腔情况,术后刺激发情期并对对明显有发情期的动物进行人工胚胎移植;3.选用性成熟、月经规律的雌性猕猴2只,根据血型和体重选取供体1例和受体1例(n=1),将切除的供体子宫原位移植到受体盆腔,术后1个月复查超声观察移植子宫情况;并将切除的受体子宫在冷保存前快速取部分宫角组织作为对照组,剩余组织使用4℃CUW (university of wisconsin)器官保存液中冷保存4h和8 h作为实验组,固定后的标本使用电镜及光镜观察子宫肌层组织不同冷缺血保存时限的病理改变;4.收集2005年12月-2015年12月在解放军总医院、武警总医院因先天性无子宫、无阴道行阴道成形术住院治疗的育龄期MRKH(mayer-rokitansky-kuster-hauser)综合征患者共31例,多次电话随访这类患者实施子宫移植的态度和意愿,调查MRKH综合征患者对子宫移植、收养、代孕的看法和选择。结果:1.5例小型猪异体子宫移植中无1例出现术中、术后并发症,髂血管吻合后均可触及血管搏动,5例受体中4例存活3个月以上,术后经腹超声于移植子宫旁及髂血管旁均可见动脉血流信号,UTx5术后第3天死亡,死亡原因为急性排斥或感染引起的子宫坏死,总的子宫移植成功率100%,受体长期存活率80%, UTx2术后3个月出现发情期并行人工胚胎移植,21天后行超声未发现妊娠;2.1例猕猴异体子宫移植术中髂血管吻合成功,术后动物存活28天,死亡原因为慢性衰竭。与对照组相比,猕猴子宫肌层组织在电镜下,使用4℃CUW器官保存液冷保存4h后未见明显改变,冷保存8h后发现核膜肿胀,核分裂,线粒体肿胀;光镜下,冷保存4h和8h后的子宫肌组织均未发现明显改变;3.31例育龄期MRKH综合征患者,有子宫移植意愿:6例,19.3%,有意愿但需要考虑或讨论的:18例,58%,无子宫移植意愿:4例,12.9%,没有表达观点:3例,9.6%; 18例需要考虑或讨论的问题:经济6例,33.3%、自己和供体可能存在的手术风险5例,27.7%、术后免疫抑制剂的副作用3例,16.6%,需要家人的支持4例,22.2%,在选择拥有孩子的方式中:子宫移植8人,25.8%,代孕 6 人,19.3%,收养 12 人,38.7%,其他 5 人,16.1%。结论:1.小型猪作为异体子宫移植活体捐赠手术是可行的,仅使用动脉系统作为移植血管受体能够长期存活且能够恢复发情期,术后超声监测能较好的评估移植子宫动脉血流;2.猕猴的是进行异体子宫移植最合适的动物模型,且子宫肌层组织至少能够耐受UW器官保存液冷保存4小时;3.育龄期MRKH综合征患者与代孕相比更愿意选择子宫移植技术来拥有一个孩子。
[Abstract]:Background: complete uterine factor infertility (abso1ute uterine factor-inferti1ity AUFI) is a major type of female infertility, which has been considered untreatable, mainly because of this type of female congenital absence of uterus such as MRKH (Mayer-Rokitansky-Kuster-Hauser) syndrome or early due to cervical cancer, obstetric hemorrhage Women of childbearing age who are forced to remove the uterus and cause no reproductive function because of severe intrauterine adhesions. The purpose of uterus transp1antation-UTx is to transplant a functional uterus and produce a child for a female with congenital or acquired factors that cause uterine deformity or uterine malformation. In order to achieve safer pregnancies and live births, a large number of animal experiments are needed to provide technical support. At home, large animal experimental studies of allogeneic uterine transplants have been at the exploratory stage, and there are few related reports. In addition, in the clinical field, the attitude and willingness of women with childbearing age subuterine factors to UTx are not known. Objective: 1. to explore the feasibility and safety of living donor donation technique for small pig uterus allograft, observe the long-term survival and reproductive function recovery of the animals after the uterus transplantation; 2. explore the surgical techniques for the living donor donation of the monkey allograft, evaluate the time limit for the cold ischemia injury in the macaque myometrium group, and 3. investigate the MRKH synthesis The patient's attitude and willingness to transplant the uterus to provide clinical data and volunteers for the next step of human uterine transplantation. 1. small pigs with 10 female and menstrual regularities were used as donors and 5 heads (n=5) based on a parent homologous (SIB) and body weight as a donor (n=5). The uterus and the main blood vessels were obtained from the pelvic cavity of the living body, using 4 centigrade C The organ preservation solution was cryopreserved and perfused to the isolated organs, then the donor uterus was transplanted in situ to the recipient pelvic cavity, and the transplanted iliac artery was anastomosed with the recipient external iliac artery end to side. 2. after the operation, 5 cases were treated with triple immunosuppressive agents, and the arterial blood flow signals and pelvic cavity were observed by Doppler ultrasound at 1 and 1 months after the operation. 2 months after the operation, the uterus and the pelvic cavity were observed two times under general anesthesia. After the operation, the oestrus was stimulated and the artificial embryos were transplanted to the apparently estrous animals. 3. female rhesus monkeys with sexual maturity and menstruation were selected to select the donor 1 cases and 1 recipients (n=1) according to the blood type and body weight, and the original displacement of the donor uterus was removed. The transplanting uterus was observed after 1 months after 1 months, and the uterus of the resected receptor uterus was taken as the control group quickly before the cold preservation. The remaining tissues were treated as the experimental group with cold preservation 4H and 8 h in the 4 CUW (University of Wisconsin) organ preservation solution. The specimens after the fixation were used by electron microscopy and light microscopy. Observe the pathological changes of different cold ischemic preservation time limits in the uterine myometrium; 4. collect 31 cases of childbearing age MRKH (Mayer-Rokitansky-Kuster-Hauser) syndrome in the General Hospital of PLA in December 2005 -2015, at the General Hospital of the PLA, because of congenital no uterus and vaginal vaginoplasty. The attitudes and wishes of the uterus transplantation were carried out to investigate the views and choices of MRKH syndrome patients for uterine transplantation, adoption, and surrogate. Results: there were no 1 cases in 1.5 cases of miniature pig allograft, postoperative complications, vascular pulsation after iliac vascular anastomosis, 4 cases in 5 patients survived more than 3 months, and postoperatively transabdominal ultrasound in transplantation The arterial blood flow signals were observed at the paranoal and iliac vessels. UTx5 died third days after the operation. The cause of death was acute rejection or infection induced uterine necrosis. The total success rate of the uterus was 100%, the long-term survival rate of the receptor was 80%. The oestrus parallel artificial embryo transfer was found at 3 months after the operation, and the pregnancy was not detected by ultrasound in 21 days; 2.1 cases of macaque allograft were not found. The iliac vascular anastomosis was successful during the operation of the uterus transplantation. The postoperative animal survived 28 days and the cause of death was chronic failure. Compared with the control group, the myometrium of the macaque was not significantly changed after the cryopreservation of 4H under the electron microscope. After the cold preservation of 8h, the swelling of the nuclear membrane, the mitosis, the swelling of mitochondria, and the cold preservation of 4H and 8h after the cold preservation were found. No significant changes were found in the myometrium of the uterus; 3.31 cases of MRKH syndrome in the childbearing age had the intention of transplanting uterus: 6 cases, 19.3%, willing but need to be considered or discussed: 18 cases, 58%, no uterine transplant Willingness: 4 cases, 12.9%, no expression of Views: 3 cases, 9.6%; 18 cases need to be considered or discussed: the economy 6, 33.3%, oneself, and donor may deposit 5 cases of operation risk, 27.7%, side effects of immunosuppressants after operation, 3 cases, 16.6%, 4 cases of family support, 22.2% in the choice of children with 8, 25.8%, 6, 19.3%, 12, 38.7%, and other 5, 16.1%. conclusion: it is feasible to operate as a living donor for allogeneic uterine transplantation. The use of the arterial system as a transplant recipient can survive for a long time and can restore the oestrus. The postoperative ultrasound monitoring can better evaluate the blood flow of the transplanted uterine artery; 2. the macaque is the most suitable animal model for the allograft of the uterus, and the myometrium of the uterus can tolerate at least 4 hours of UW organ preservation and 4 hours; 3. of the childbearing age. The combined patients are more willing to choose uterine transplant technology to have one child than surrogacy.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R-332;R711.6

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