子宫内膜异位症影响妊娠的相关因素分析及动物模型的建立

发布时间：2018-06-06 06:29

本文选题：子宫内膜异位症 + 体外受精-胚胎移植　；参考：《安徽医科大学》2017年硕士论文

【摘要】：1目的探索子宫内膜异位症(EMS)对不孕患者经体外受精-胚胎移植(IVF-ET)治疗后妊娠结局的影响及妊娠相关因素的分析;并以小鼠为实验对象,通过手术造模,探索子宫内膜异位症小鼠模型成模状态最佳的时间,为以后相关研究提供参考,并为研究子宫内膜异位症对小鼠胚胎的影响做准备。2方法2.1、临床研究:选择自2012年1月至2016年6月在我中心行IVF-ET治疗的46例子宫内膜异位症合并不孕患者的55个取卵周期作为内异症组(EMS组);选择同期行IVF-ET治疗的126例非子宫内膜异位症患者的156个周期作为对照组(非EMS组),平衡两组患者的一般情况后,从CA-125水平、窦卵泡数(AFC)、促排卵药物启动量/总量、HCG日内膜厚度/类型、HCG日大卵泡数(直径≥14mm卵泡)、HCG日雌二醇(estrogen,E2)、HCG日每大卵泡数E2、获卵数、受精方式、MII数、正常受精数/卵裂数、优胚数、正常受精率、卵裂率、优胚率、胚胎着床率、生化妊娠率、临床妊娠率、早期流产率、活产率、周期取消率等方面进行回顾性分析,了解EMS对EMS相关性不孕患者的实验室及临床结局的影响;再将两组临床资料按临床结局分别分为妊娠组、未妊娠组,进一步明确EMS组和非EMS组中与妊娠结局相关的因素。2.2、实验研究:1)选择6周龄雌性无特殊病原体(specific pathogen free,SPF)级KM(昆明)小鼠33只,随机分为三组,每组11只。2)适应新环境1周后,手术取出小鼠左侧子宫角,均分为4段,剪开子宫腔,得到四块子宫组织。其中两块组织以内膜面朝向腹壁的方向分别缝合在切口两侧腹壁上,一块组织剪碎后注入小鼠腹腔,剩余一块组织送病理检查,以确认所取组织来源于小鼠子宫;三组小鼠均采用此方法建立子宫内膜异位症模型。3)于手术后2周、3周、4周分别处死三组小鼠,观察内异灶形态,并进行如下统计计算:(1)比较各组小鼠的术后存活率、不同部位病灶形成率、异位灶囊泡形成率及模型成功率;(2)组内比较腹壁、网膜、肠壁三个部位组织碎片的种植成功率;(3)组内比较缝合、碎片种植、缝合+碎片种植三种建模方式的成功率;(4)比较三组小鼠异位病灶的平均体积。3结果3.1、临床研究:EMS组CA-125水平、周期取消率高于对照组(P0.05);基础窦卵泡数(AFC)、HCG日大卵泡数、获卵数、MII数、2PN数、卵裂数、优胚数、优胚率低于对照组(P0.05);HCG日大卵泡数与EMS患者妊娠失败的相关回归系数小于0(P0.05)。3.2、实验研究:本实验采用KM小鼠进行EMS的建模,参与本实验的33只小鼠中,术后存活28只。(1)三组小鼠术前未区分动情周期,但术后存活率、各部位病灶形成率、囊泡形成率及模型成功率比较,差异均无统计学意义(P0.05);(2)三组的组内腹壁、网膜、肠壁三个部位内膜碎片种植成功率的比较,差异均无统计学意义(P0.05);(3)三组中组内缝合、碎片种植、缝合+碎片种植三种方式建模方式成功率的比较,其中仅术后4周组中碎片种植比缝合、缝合+碎片种植两种建模方式的成功率低,差异有统计学意义(P0.05),后两者方式的建模成功率均为100%;(4)三组小鼠异位病灶平均体积比较结果显示,术后2周小鼠异位病灶体积小于术后4周小鼠异位病灶体积(4.00±2.85mm3 Vs.25.64±15.93mm3,P=0.001),差异有显著性统计学意义。4结论4.1、临床研究:子宫内膜异位症合并不孕患者卵巢储备功能下降,胚胎质量下降,更容易取消周期。但其卵子成熟率、正常受精率、卵裂率及临床结局与非内异症不孕患者相比无明显差别;HCG日大卵泡数是EMS患者妊娠的保护因素。4.2、实验研究:1)术后2周EMS模型已成功,且造模前无需统一动情周期;2)子宫组织碎片在腹壁、大网膜、肠壁的种植率无差异;3)术后2-3周行下一步研究的,可采用子宫组织碎片腹腔种植法建模;术后4周行下一步研究的,可采用子宫片段腹壁缝合法建模;4)造模术后4周,异位灶平均体积最大,EMS模型效果最佳。
[Abstract]:1 Objective To explore the effect of endometriosis (EMS) on pregnancy outcome after the treatment of in vitro fertilization and embryo transfer (IVF-ET) and the analysis of pregnancy related factors. In order to study the effect of endometriosis on mouse embryos,.2 method 2.1 was prepared. Clinical study: 55 oval cycles in 46 cases of endometriosis with infertility from January 2012 to June 2016 were selected as endometriosis group (group EMS), and 126 non endometrium were selected for IVF-ET treatment at the same time. 156 cycles of heterotopic patients (non EMS group), balance two groups of patients, from the level of CA-125, the number of sinus follicles (AFC), the amount of oviposit promoter / total, HCG diurnal intima thickness / type, HCG day large follicle number (diameter > 14mm follicle), HCG day estradiol (estrogen, E2), HCG day per large follicle number E2, the number of eggs, fertilized square Formula, MII number, normal fertilization number / cleavage number, optimal embryo number, normal fertilization rate, cleavage rate, excellent embryo rate, embryo implantation rate, biochemical pregnancy rate, clinical pregnancy rate, early abortion rate, survival rate, cycle cancellation rate, etc., to understand the effect of EMS on the laboratory and clinical outcome of EMS related infertile patients; and then two groups of clinical funds According to the clinical outcome, the material was divided into pregnancy group and non pregnancy group, and the factors related to pregnancy outcome in group EMS and non EMS group were further clarified.2.2, experimental study: 1) 33 mice of 6 weeks old female without special pathogen (specific pathogen free, SPF) KM (Kunming) mice were selected, and were divided into three groups, each group of 11.2) adapted to the new environment for 1 weeks and the operation was removed. The left corner of the uterus of the mice was divided into 4 segments, and the uterine cavity was cut open and four uterine tissues were cut. Among them, two tissues were sutured on both sides of the abdominal wall in the direction of the endometrium toward the abdominal wall. One tissue was cut into the abdominal cavity, and the rest of the tissue was sent to the pathological examination to confirm that the tissues were derived from the uterus of mice; the three groups were all of the mice. Using this method to establish endometriosis model.3), three groups of mice were killed at 2 weeks, 3 weeks and 4 weeks after the operation. The morphology of the endometrium was observed and the following statistical calculations were performed. (1) the survival rate, the rate of focal lesion formation in different parts of the mice, the formation rate of the ectopic foci and the success rate of the model were compared. (2) the abdominal wall, omentum, and the wall of the intestine were compared in the group (2) The success rate of tissue fragmentation in three parts; (3) the success rate of three modeling methods: comparison suture, shard planting, suture + fragment planting; (4) the average volume of.3 in three groups of mice was 3.1. Clinical study: the CA-125 level in group EMS was higher than that of the control group (P0.05); the number of basal sinus follicles (AFC), and large follicle of HCG day Number, number of eggs, MII number, 2PN number, cleavage number, optimal embryo number, optimal embryo rate lower than that of control group (P0.05); the correlation coefficient of large follicle number and EMS patient's pregnancy failure was less than 0 (P0.05).3.2, experimental study: this experiment used KM mice to model EMS, and 28 mice survived the 33 mice in this experiment. (1) three groups of mice did not distinguish before operation. The estrous cycle, but the postoperative survival rate, the rate of lesion formation, the rate of vesicle formation and the success rate of the model were not statistically significant (P0.05). (2) there was no significant difference in the success rate of the internal abdominal wall, omentum and the three parts of the intestinal wall of the three groups (P0.05); (3) the three groups of sutures, shards, suture + The success rates of three modeling methods were compared, in which only 4 weeks after operation, the success rate of two kinds of modeling methods, including shard planting ratio suture, suture + fragment planting, was statistically significant (P0.05), and the success rate of the latter two models was 100%. (4) the average volume comparison of the three groups of heterotopic lesions showed that 2 weeks after the operation. The volume of heterotopic focus in mice was less than 4 weeks after the operation (4 + 2.85mm3 Vs.25.64 + 15.93mm3, P=0.001). The difference was statistically significant.4 conclusion 4.1. Clinical study: the ovarian reserve function of endometriosis combined with infertility decreased, the quality of embryo decreased and the cycle was easier to cancel, but the rate of egg maturation was normal. There was no significant difference in fertilization rate, cleavage rate and clinical outcome compared with infertile infertility; HCG day large follicle number was a protective factor for pregnancy in EMS patients.4.2, experimental study: 1) 2 weeks after the operation, EMS model had been successful, and no unified estrous cycle was needed before modeling; 2) the implantation rate of uterine tissue fragments on abdominal wall, omentum, and intestinal wall was no difference; 3) 2-3 after operation. The next step of study in the next week can be modeled by the method of intraperitoneal implantation of uterine tissue fragments. 4 weeks after the operation, the next step of the study can be modeled by the uterine segment abdominal suture; 4) the average volume of the heterotopic focus is the largest after 4 weeks, and the EMS model is the best.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.71;R-332

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