低强度超声对大鼠早孕药物流产后子宫复旧的影响及其机制的初步研究

发布时间：2018-04-11 03:10

本文选题：低强度超声 + 子宫复旧　；参考：《重庆医科大学》2014年硕士论文

【摘要】：目的 1.观察低强度超声对早孕药流子宫的收缩作用，分析其对子宫复旧的影响。 2.探讨低强度超声促进早孕大鼠药物流产后子宫复旧的可能机制，为低强度超声促进产后子宫复旧提供理论依据。1 方法 1.随机选择3月龄以内的雌性普通级远交群大鼠（SpragueDawley，SD）55只，雄性普通级SD大鼠5只，按照雌雄2:1的比例夜晚（20:00）合笼，12小时后（次日8:00）观察阴道栓，然后进行阴道涂片于显微镜下观察精子。以显微镜发现精子或明确查见阴道栓的当天为妊娠第1天。若未发现，雌雄大鼠分笼喂养7天后，按前法再次合笼至妊娠，妊娠第7天，依次灌胃米非司酮16.6mg/kg（8:00）与米索前列醇0.1mg/kg（18:00），建立30只大鼠早孕药物完全流产模型。 2.将建模成功的30只大鼠采用完全随机分组法分组到超声辐照组和假辐照组各15只，超声辐照组：药流后第一天开始，使用超声波子宫复旧仪照射子宫覆盖区域（声强=2W/cm2，频率=0.8MHz）。每日1次，每次30分钟，连续5天；假辐照组：从药流后第一天开始使用超声波子宫复旧仪照射子宫覆盖区域（声强=0W/cm2，频率=0MHz）。每日1次，每次30分钟，连续5天。 3.孕囊排出后，用眼科镊于大鼠阴道内置入定量棉球一个，每2~6小时取出血棉，检测其出血量，并记录出血时间。 4.治疗完成后次日，取大鼠左侧子宫组织黄豆大小3块，采用酶联免疫吸附法（ELISA）、化学检测法、免疫组织化学法（IHC）分别检测内皮素-1（endothelin-1，ET-1）、一氧化氮（nitrogen monoxide，NO）和缩宫素受体（oxytocin receptor，OTR）。结果 1.受孕成功率为41.7%（40/96）。三种检查方法，灵敏度阴栓+精子检查（0.9000）最高，与精子检查（0.8750）相差不大，阴栓检查灵敏度（0.7000）最低，特异度三种方法相差不大；阴栓检查假阴性率高达30.00%，与精子检查（12.50%）和联合检查相比（10.00%）差异均有统计学意义（*P＜0.05），三种检测受孕方法比较假阳性率无显著性差异（P＞0.05）。药物流产成功率为88.24%（30/34）。 2.肉眼观：假辐照组，子宫已不见孕囊窝，但较正常大鼠子宫粗约2mm~3mm，表面充血明显，呈樱红色；超声辐照组，大鼠子宫大体观已基本恢复至未孕状态。病理：超声辐照组，子宫肌纤维细长，排列规则，内环外纵；假辐照组，子宫肌纤维轻度增生肥大，排列稍紊乱，子宫肌层整体较超声辐照组厚，子宫腔较大。 3.阴道出血量、出血时间的比较，超声辐照组的大鼠阴道出血量明显少于假辐照组，差异有统计学意义（*P＜0.05）；但阴道出血时间超声辐照组和假辐照组之间没有显著性差异（P＞0.05） 4.超声辐照组的大鼠子宫组织匀浆中ET-1的含量明显高于假辐照组，具有显著的统计学意义（*P＜0.05），，而超声辐照组大鼠子宫匀浆中NO的含量较低，但没有明显的统计学意义（P＞0.05）。 5.超声辐照组OTR表达多为强阳性，假辐照组OTR表达多数为弱阳性；无论子宫肌层还是内膜层超声辐照组强阳性表达率均高于假辐照组，且差异均有显著统计学意义（*P＜0.05）。结论 1.精子检查法操作简便，方法易于掌握，灵敏度高，为最佳的判断大鼠雌鼠受孕的方法；米非司酮16.6mg/kg＋米索前列醇0.1mg/kg剂量组建立完全药流模型成功率高，可有效的运用于大鼠完全流产模型的建立。 2.声强=2W/cm2，频率=0.8MHz的低强度超声作用于大鼠完全流产子宫，可促进早孕大鼠流产后的子宫复旧。 3.增加子宫组织ET-1的含量，对ET-1和NO平衡的调节可能是低强度超声促进子宫收缩的重要途径之一。 4.低强度超声促进子宫平滑肌收缩的机制与增加子宫肌和内膜细胞的OTR表达有关。
[Abstract]:objective
1. to observe the contraction effect of low intensity ultrasound on uterine pregnancy abortion, analyzes its influence on uterine involution.
2. to explore the mechanism of low intensity ultrasound can promote uterine involution in early pregnancy rats after drug abortion, and provide a theoretical basis for the.1 of low intensity ultrasound in promoting postpartum uterine involution
Method
The female general level within 1. randomly selected 3 month old outbred rats (SpragueDawley, SD) 55, male SD 5 rats, according to the proportion of male and female 2:1 night (20:00) cage, after 12 hour (8:00 the next day) to observe vaginal suppository, and vaginal smear under a microscope to observe sperm. In order to find microscope sperm or definitely check the day to see vaginal suppository for pregnancy first days. If not found, male and female rats were divided into the cage for 7 days, according to the law again before cage to pregnancy, pregnancy seventh days, followed by intragastric administration of mifepristone and misoprostol 0.1mg/ 16.6mg/kg (8:00) kg (18:00), established in 30 rats early pregnancy abortion model completely.
30 rats 2. successful modeling were randomly grouped into ultrasound group and sham irradiation group with 15 rats in each group, ultrasound group: the first day after medical abortion, the use of ultrasonic irradiation of uterine involution of uterus coverage area (intensity of =2W/cm2, frequency =0.8MHz). 1 times a day, every 30 minutes, continuous 5 days; sham irradiation group: from the first day of the medical abortion after the start of the use of ultrasonic irradiation of uterine involution of uterus coverage area (intensity of =0W/cm2, frequency =0MHz). 1 times a day, every 30 minutes, for 5 consecutive days.
After 3. gestation sac was discharged, a quantitative cotton ball was inserted into the vagina of the rat with an eye tweezers. The blood cotton was removed every 2~6 hours, and the amount of bleeding was detected and the bleeding time was recorded.
The 4. day after treatment, the size of soybean left uterine tissue of rats 3, using enzyme-linked immunosorbent assay (ELISA), chemical assay, immunohistochemistry (IHC) were used to detect endothelin -1 (endothelin-1, ET-1), nitric oxide (nitrogen monoxide, NO) and oxytocin receptor (oxytocin receptor, OTR).
Result
The success rate of pregnancy was 41.7% (40/96) 1.. Three methods, the sensitivity of the pessary + sperm examination (0.9000) and the highest sperm examination (0.8750) or less, the vaginal examination (0.7000) the lowest sensitivity, specificity three methods had little difference; vaginal plug the false negative rate is as high as 30%, and sperm check (12.50%) compared with the joint inspection (10%) the differences were statistically significant (*P < 0.05), there was no significant difference between the three methods for detection of pregnancy compared with the false positive rate (P > 0.05). The success rate of drug abortion was 88.24% (30/34).
2. eye view: sham irradiation group, the uterus has not seen the pregnancy sac fossa, but compared with the normal rat uterus about 2mm~3mm thickness, surface congestionsignificantly, was cherry red; ultrasonic irradiation group, rat uterus concept has been basically restored to the nonpregnant state. Pathology: ultrasound irradiation group, uterine muscle fiber slender, regular arrangement the inner ring, outer longitudinal; sham irradiation group, mild uterine muscle fiber hypertrophy, arranged a little disorder. The myometrium overall than the ultrasound group thickness of uterine cavity is larger.
3., compared with vaginal bleeding and bleeding time, the amount of vaginal bleeding in the ultrasonic irradiation group was significantly less than that in the sham irradiation group (*P < 0.05), but there was no significant difference in vaginal bleeding time between the ultrasonic irradiation group and the sham irradiation group (P > 0.05).
4. the content of ET-1 in the uterine tissue homogenate of the ultrasonic irradiation group was significantly higher than that of the sham irradiation group (*P < 0.05), while the content of NO in the uterine homogenate of the ultrasonic irradiation group was lower, but there was no significant statistical difference (P > 0.05).
5., the expression of OTR in ultrasonic irradiation group was strongly positive. Most of the OTR expression in sham irradiated group was weakly positive. The expression of strong positive in ultrasound and irradiation group was higher than that in sham irradiation group, and the difference was statistically significant (*P < 0.05).
conclusion
1. sperm examination method is simple, the method is easy to grasp, high sensitivity, as the best way of female pregnancy rats were assessed by mifepristone misoprostol; 16.6mg/kg + 0.1mg/kg group to establish complete abortion model with high success rate and can be effectively applied to establish the rat model of complete abortion.
2. sound intensity =2W/cm2, low intensity ultrasound frequency =0.8MHz in rat uterus can promote complete abortion, early pregnancy rats after abortion uterine involution.
3. increasing the content of ET-1 in the uterus and regulating the balance of ET-1 and NO may be one of the important ways to promote the contraction of the uterus by low intensity ultrasound.
4. the mechanism of promoting contraction of uterine smooth muscle by low intensity ultrasound is related to the increase of OTR expression in the endometrium and endometrium cells.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R445.1;R714.63

【参考文献】