裸鼠人来源性子宫腺肌病造模方法的比较

目的通过不同的造模方法建立裸鼠人来源性子宫腺肌病模型,筛选出理想的造模方法,为研究子宫腺肌病发病机制及相关治疗方案提供理想的动物模型。方法将80只雌性裸鼠随机分为腹腔种植组、皮下种植组、腹腔注射组和皮下注射组,每组20只。各模型组分别采用不同造模方法建立裸鼠人来源性子宫腺肌病模型,术后4周观察移植物的体积及组织形态学。结果腹腔种植组建模成功率95%,皮下种植组建模成功率45%,腹腔注射法及皮下注射法成模率0%。结论腹腔种植法建立子宫腺肌病裸鼠模型成模率高,稳定性良好,是一种理想的人来源性子宫腺肌病动物模型。     

口服他莫昔芬法建立ICR小鼠子宫腺肌病模型

目的使用口服他莫昔芬法建立ICR小鼠子宫腺肌病模型,并检测其病灶特征、动情周期、血管生成、子宫炎症等变化,以介绍和评价这一动物模型。方法新生ICR小鼠（15只）连续4 d滴喂他莫昔芬,并与同龄对照小鼠（15只）分别于42、85-95、135-145日龄处死,使用苏木素-伊红染色检测子宫病理改变;阴道脱落细胞法检测动情周期变化;免疫组化补体31（CD31）染色计算子宫微血管的密度、直径及所占面积比;逆转录-聚合酶链反应（RT-PCR）检测子宫缓激肽受体、神经激肽受体的基因表达。结果使用口服他莫昔芬法建立ICR小鼠子宫腺肌病模型的造模率为100%,且疾病严重程度随病程进展。部分给药小鼠可出现动情周期紊乱。85-95及135-145日龄给药小鼠子宫肌层微血管密度和面积比均高于对照小鼠（P〈0.05）。135-145日龄给药小鼠子宫缓激肽受体、神经激肽受体的基因表达较对照组明显升高（P〈0.05）。结论口服他莫昔芬法可方便、高效的建立腺肌病小鼠模型,出现腺肌病相关的血管生成、炎症状态、疼痛相关受体表达增高等特征,是研究腺肌病发生、发展的良好模型。     

寒冷对雌性C57BL/6小鼠动情周期的影响*

目的:研究寒冷对雌性C57BL/6小鼠动情周期的影响。方法:12只雌性小鼠随机分为对照组、低温组,每组6只;低温组每天4℃暴露4 h,每天阴道涂片法观察小鼠动情状况,对照组饲养于常温动物房;每2 d称量体重,2周后心脏取血、子宫和卵巢,检测小鼠血清E2、FSH、LH、Prl、P水平,进行子宫、卵巢的组织病理学检查。结果:与对照组比较,低温组小鼠体重无显著性差异(P＞0.05),小鼠子宫脏器系数明显较低、动情间期明显延长(P＜0.01),血清FSH显著升高、Prl显著降低(P＜0.01),小鼠子宫腺管扩张,卵巢卵泡数量明显减少。结论:寒冷可使雌性C57BL/6小鼠动情周期延长,进而可能影响生殖功能。     

低剂量双酚A灌胃对去卵巢小鼠子宫的增生作用

目的观察小鼠经消化道途径给予低剂量环境内分泌干扰物双酚A对子宫的作用及机制。方法雌性ICR小鼠切除双侧卵巢,恢复性饲养1周,剔除异常个体后,50只动物随机分成5组,每组10只动物,依次皮下注射(sc)10.0μg/(kg·d)17β-雌二醇,灌胃(ig)0、20.0、60.0和180.0μg/(kg·d)BPA,连续7d。动物每3d称量体重一次,于最后一次给药24h后,阴道涂片检测角化上皮细胞,颈椎脱臼处死动物,取子宫,称其干湿重,计算子宫脏器指数,组织切片,HE染色后利用图像分析系统测量子宫上皮高度和宫腔面积。免疫组化法分析子宫雌激素受体(ER)和孕激素受体(PR)表达。结果①与对照组相比,雌二醇和BPA低剂量组动物体重增加,BPA中高剂量组动物体重无明显差异。②阴道涂片结果显示,处于动情期动物数分别为:对照组0/10,雌二醇组10/10,BPA低剂量组2/10,BPA中剂量组1/10,BPA高剂量组0/10;③BPA低剂量组动物子宫湿重和含水量较对照组增加,脏器系数增大,BPA中高剂量组子宫湿重和脏器系数较对照组降低明显(P0.05);④病理组织学及显微图像分析结果显示,BPA低剂量组动物子宫腔面积较对照组增大,中高剂量组子宫腔面积明显减小(P0.01);低剂量组动物子宫上皮高度较对照组增高(P0.01),中高剂量组子宫上皮高度降低明显(P0.01)。⑤免疫组化结果显示,低剂量BPA能增强小鼠子宫ER和PR表达,但随剂量增加,ER表达下降。结论BPA经消化道途径给予ICR小鼠,人环境暴露剂量下具有雌激素活性,但雌激素活性随BPA剂量增大呈现下降趋势。     

注射法小鼠皮下子宫内膜异位症模型及组织学观察

目的 建立ICR小鼠皮下子宫内膜异位症新的模型及组织学观察.方法 以同系小鼠作为供体,取子宫内膜分别采用皮下注射法和皮下植入法对2组共24只小鼠行皮下造模手术.3周后测量皮下种植结节大小,并处死半数造模成功小鼠,取子宫和异位结节行HE染色组织学研究.6周后,处死剩余小鼠,并再次测量皮下结节,取子宫和异位结节行HE染色组织学研究.结果 皮下植入法和皮下注射法成功率分别为33.33%和83.33%(P<0.01);生长良好的囊肿囊内壁有内膜上皮细胞生长,有与正位子宫内膜接近同步的发情周期改变;病灶随移植时间的推延先逐渐增大之后又有缩小趋势(P>0.05).结论 将ICR小鼠子宫内膜注射到同系小鼠腹部皮下形成子宫内膜异位病理模型,成功率高,且简单可行,异位内膜随动情周期有周期性改变,生理特性与正位子宫内膜基本相同,应用于子宫内膜异位症疾病和药物研究既经济又稳定可靠.     

NGF 及其受体TrkA 在子宫腺肌病中的表达及与痛经的关系

目的:研究NGF及其受体TrkA在子宫腺肌病患者的异位内膜与在位内膜组织的表达情况及与痛经的关系。方法:采用免疫组化MaxVision法检测子宫腺肌病异位内膜(30例)、在位内膜(30例)、正常子宫内膜(19例)标本中NGF、TrkA蛋白的表达,分析其表达差异及与痛经的关系。结果:①子宫腺肌病异位内膜组NGF、TrkA表达显著高于正常内膜组(P<0.01),在位内膜组NGF、TrkA表达显著高于正常内膜组(P<0.01),子宫腺肌病异位内膜组NGF、TrkA表达与在位内膜组无显著差异。②子宫腺肌病异位内膜组NGF、TrkA表达与痛经强度评分呈正相关(相关系数r=0.637,P=0.000;r=0.662,P=0.000)。结论:NGF及其受体TrkA在子宫腺肌病中高表达可能参与子宫腺肌病发病机制,而且可能与痛经有关。     

NGF及其受体TrkA在子宫腺肌病中的表达及与痛经的关系

目的：研究NGF及其受体TrkA在子宫腺肌病患者的异位内膜与在位内膜组织的表达情况及与痛经的关系。方法：采用免疫组化MaxVision法检测子宫腺肌病异位内膜（30例）、在位内膜（30例）、正常子宫内膜（19例）标本中NGF、TrkA蛋白的表达,分析其表达差异及与痛经的关系。结果：①子宫腺肌病异位内膜组NGF、TrkA表达显著高于正常内膜组（P〈0.01）,在位内膜组NGF、TrkA表达显著高于正常内膜组（P〈0.01）,子宫腺肌病异位内膜组NGF、TrkA表达与在位内膜组无显著差异。②子宫腺肌病异位内膜组NGF、TrkA表达与痛经强度评分呈正相关（相关系数r=0.637,P=0.000;r=0.662,P=0.000）。结论：NGF及其受体TrkA在子宫腺肌病中高表达可能参与子宫腺肌病发病机制,而且可能与痛经有关。     

NGF蛋白及前列腺F2α受体与子宫腺肌病痛经的相关性研究

目的:研究NGF蛋白及前列腺F2α受体(PTGFR)与子宫腺肌病痛经的相关性。方法:选择我院2016年7月～2017年7月收治的36例子宫腺肌病痛经患者,按视觉模拟评分法(VAS)将痛经程度分为11例轻度组、14例中度组、11例重度组。同期选择36例月经正常者作为对照组。比较各组血清NGF蛋白和血浆PTGFR水平,分析二者和痛经评分之间的相关性及其单独和联合检测时诊断子宫腺肌病痛经的敏感性、特异性及受试者工作特征(ROC)曲线下面积。结果:子宫腺肌病痛经组血清NGF蛋白和血浆PTGFR水平均显著高于对照组(P0.05)。轻度痛经组血清NGF蛋白及血浆PTGFR水平均显著低于中度组和重度组(P0.05)。血清NGF蛋白和血浆PTGFR水平和子宫腺肌病痛经评分均呈显著正相关,r分别为0.812,0.884(P0.05)。ROC曲线分析结果显示血清NGF蛋白联合血浆PTGFR蛋白检测诊断子宫腺肌病痛经的ROC曲线下面积明显大于血清NFG蛋白及血浆PTGFR水平单独检测(P0.05)。结论:NGF蛋白和PTGFR可能参与了子宫腺肌病痛经的发生和发展,二者联合检测有助于诊断子宫腺肌病痛经。血清NGF蛋白水平及血浆PTGFR蛋白水平和子宫腺肌病痛经程度呈正相关。     

卵巢组织异体移植对去势模型大鼠骨质疏松的治疗作用

目的:研究卵巢组织异体移植对去势模型大鼠骨质疏松的治疗作用及其机制。方法:将40只SD雌性大鼠随机分为假手术组、去卵巢模型组,雌激素组和卵巢组织块移植组。采用背部定位切除双侧卵巢,建立去势大鼠模型。造模的同时,将同种异体卵巢组织块植入去卵巢大鼠背部肌肉层以建立卵巢移植模型。雌激素组大鼠按雌激素0.06 mg/kg灌胃进行替代治疗3个月,其他各组均给予等量生理盐水。实验结束后,称量体重和子宫湿重,计算子宫重量指数;HE染色光镜观察大鼠阴道上皮细胞和股骨细胞形态;骨密度仪法检测大鼠全身、股骨和胫骨近端的骨密度;放射免疫法检测血清雌激素(E2)水平。结果:与假手术组相比,模型组大鼠子宫重量指数明显下降(P0.01);血清雌激素水平显著降低(P0.01);阴道上皮细胞较小,无角质化;股骨中的骨小梁数目显著减少,骨吸收明显,呈典型的"岛屿状",骨髓腔增大,髓腔中基质细胞极少;双能X线显示,模型大鼠全身、股骨和胫骨的骨密度均显著减少(P均0.01);与模型组相比,雌激素组和移植组大鼠的子宫重量指数增加明显(P0.01,P0.01);两组大鼠的血清雌激素水平上升显著(P0.01,P0.01);病理切片显示,雌激素组和移植组大鼠的阴道上皮细胞大多呈角质化;雌激素组大鼠皮质骨增厚,骨小梁粗壮,数量多,排列较整齐,骨小梁间距较小,且骨髓腔缩小,骨基质细胞明显增多;移植组大鼠皮质骨面积较大,骨小梁数量和面积增加明显,骨小梁间距小,排列规则,连接增多,骨基质细胞分布均匀。结论:卵巢组织移植可能是通过与靶肌肉组织产生新生血管连接,不断分泌内源性雌激素,后者通过结合子宫、阴道和股骨组织中靶受体-雌激素受体,进而维持子宫和阴道等靶组织的形态与功能,并抑制骨质组织矿物质的丢失,从而达到改善或促进雌性大鼠内分泌功能的目的。     

选择性雌激素受体β激动剂对子宫腺肌病模型小鼠孕激素受体和胰岛素样生长因子-1的影响

摘要 目的：探讨选择性雌激素β受体（Estrogen Receptor beta,ERβ）激动剂对子宫腺肌病模型小鼠孕激素受体和胰岛素样生长因子-1的影响。方法：子宫腺肌病小鼠(n=30)随机平分为三组-模型组、孕三烯酮组与选择性ERβ激动剂组。模型组每日灌胃蒸馏水0.4 mL/20 g体重,孕三烯酮组每日灌胃孕三烯酮水溶液0.008 mg/20 g体重,选择性ERβ激动剂组每日灌胃WAY-32255水溶液0.008 mg/20 g体重,连续口服14 d。结果：孕三烯酮组与选择性ERβ激动剂组治疗第7 d与第14 d的小鼠体重高于模型组（P<0.05）,选择性ERβ激动剂组高于孕三烯酮组（P<0.05）。孕三烯酮组与选择性ERβ激动剂组治疗第7 d与第14 d的子宫病理评分低于模型组（P<0.05）,选择性ERβ激动剂组低于孕三烯酮组（P<0.05）。孕三烯酮组与选择性ERβ激动剂组治疗第7 d与第14 d的血清孕激素受体（Progesterone receptor,PR）和胰岛素样生长因子（insulin like growth factor,IGF-1）含量低于模型组（P<0.05）,选择性ERβ激动剂组低于孕三烯酮组（P<0.05）。孕三烯酮组与选择性ERβ激动剂组治疗第7 d与第14 d的子宫RhoA和ROCK蛋白相对表达水平低于模型组（P<0.05）,选择性ERβ激动剂组低于孕三烯酮组（P<0.05）。结论：选择性ERβ激动剂对子宫腺肌病模型小鼠的应用能抑制血清PR和IGF-1的释放,降低RhoA和ROCK蛋白的表达,从而能改善子宫病理状况,提高小鼠体重。     

Effects of mifepristone (RU486) treatment on the development of uterine adenomyosis induced by pituitary grafting in mice

To evaluate the effects of mifepristone (RU486) on the development of uterine adenomyosis induced by pituitary grafting (PG), 3 groups of mice receiving pituitary grafts at 7 weeks of age were given RU486 in food (20 mg/kg chow) from 3-14 (RU486-3 group) or 10-14 (RU486-10 group) weeks of age, or were given no further treatment (PG control group), respectively. All the mice were killed at 14 weeks of age. The uterine weight was significantly decreased in both RU486-treated groups compared with the PG control group. The incidence of adenomyosis was also decreased significantly in both the RU486-3 group (0/10 mice) and RU486-10 group (2/10 mice) compared with the PG control group (7/9 mice). To look for vascular changes in the uterine tissues, which have been reported to be related to the development of adenomyosis, immunohistochemical staining of von Willebrand factor in the blood vessels was performed. The mean surface area and minor axis of blood vessels in the uterus were thereby found to be significantly decreased in the RU486-10 group compared to the PG control group. The results clearly indicated that RU486, a potent antiprogestin, could inhibit the genesis of uterine adenomyosis in mice, and at the same time caused shrinkage of the vascular system. As in humans, progesterone as well as the vascular system therefore appear to be important factors in the pathogenesis of uterine adenomyosis in this mouse model.     

Increase of DNA synthesis in uterine adenomyosis in mice with ectopic pituitary isograft.

Ectopic pituitary isografts (EPI) have been found to induce a high incidence of uterine adenomyosis in SHN mice. All the SHN mice given EPI in the right uterus at 40 days of age developed uterine adenomyosis, and more than 80% of mice showed the genesis of subserosal nodules, an advanced state of adenomyosis, 65 days after EPI. Activities of both thymidylate synthetase and thymidine kinase, i.e. DNA-synthesizing enzymes in de novo and salvage pathways of pyrimidine metabolism, respectively, were significantly increased in EPI-induced uterine adenomyosis to approximately 2-fold those in normal control uteri. Bromodeoxyuridine-immunoreactive cells were regarded as the cells in S phase, and the number in the endometrial epithelium and stroma in EPI-induced uterine adenomyosis was more than 1.5-fold that in normal control uteri. EPI may affect the genesis of uterine adenomyosis generally, but not locally, because there were no differences between the right uterus with EPI and the left without EPI in the incidence of adenomyosis, histology or DNA-synthesizing enzyme activities.     

Development of uterine adenomyosis after treatment with dopamine antagonists in mice

Development of uterine adenomyosis was studied in SHN mice treated with psychotherapeutic drugs, sulpiride and perphenazine, and gastroenteric drug, metoclopramide, which act as dopamine antagonists to increase prolactin release from the pituitary gland. Administration of these drugs twice daily for 40-70 or 40-90 days of age induced an elevation in serum level of prolactin. Furthermore, the treated mice showed a prolongation of metestrous plus diestrous phase and a high incidence of uterine adenomyosis compared with vehicle-treated control mice. These results indicate that hyperprolactinemia produced by continuous treatment with psychotherapeutic and gastroenteric drugs is responsible for the occurrence of irregular estrous cycles and the genesis of uterine adenomyosis in mice.     

Priming effects of novel nonsteroidal progesterone receptor modulators CP8816 and CP8863 on the development of adenomyosis in the mouse uterus

The possibility of therapeutic application of novel nonsteroidal progesterone receptor modulators CP8816 and CP8863 for preventing the development of uterine adenomyosis was investigated in mice. First priming effects of CP8816 on 17beta-estradiol (E2)-induced cell division in uterine tissues were examined. As a result, pretreatment with CP8816 or progesterone significantly suppressed the elevation of the mitotic activity in the luminal epithelial cells of mice treated with E2 later. Priming with CP8816 had little effect on the stromal cells, but progesterone priming caused an increase of stromal mitotic activity in mice treated with E2 later. To evaluate the inhibitory effect of these compounds on the development of adenomyosis induced experimentally by pituitary grafting, 7-week-old female mice were isografted with a single anterior pituitary in the uterus and divided into four groups. Two groups of mice were given daily subcutaneous injections of 1 mg of CP8816 or the vehicle alone for 6 weeks from the day after the grafting. Remaining two groups of mice were given oral administration of 1 mg of CP8863 or the vehicle only for 5 weeks starting one week after the grafting. The incidence of adenomyosis was significantly lower in the groups of mice treated with CP8816 and CP8863 than in the respective control groups. The mechanism by which CP compounds inhibited the development of adenomyosis might be related to their priming effects, i.e., their inhibitory effect on epithelial cell division and lack of effect on stromal cell division after subsequent exposure to E2.     

The induction of adenomyosis in mice by intrauterine pituitary isografts

Implantation of a single anterior pituitary into the uterine lumen induced the high incidence of adenomyosis in SHN strain of mice. The subserous nodules of the advanced state of adenomyosis appeared in larger numbers in the right uterine horns bearing the pituitary isografts than in the left ones bearing no isografts. Ovariectomy after implantation completely eliminated the occurrence of adenomyosis. Meanwhile, continuous treatments with estrogen in combination with progesterone recovered such pathological state in the ovariectomized mice receiving the pituitary isografts. The circulating levels of prolactin were consistently higher in mice given pituitary isografts than in the controls given isografts of pieces of submaxillary glands. The results indicate that, in this system, prolactin plays an important role in the genesis of adenomyosis, although the effect of ovarian sex steroids cannot be excluded.     

改良腹腔镜手术治疗子宫腺肌症的可行性及安全性研究

目的:探讨改良腹腔镜手术治疗子宫腺肌症的可行性及安全性。方法:84例子宫腺肌症患者,采用方便抽样方法选取接受改良腹腔镜子宫体楔形切除术的40例患者作为研究组,接受传统腹腔镜全子宫切除术的44例患者作为对照组,比较两组手术情况及术后康复情况。结果:两组术中出血量、手术时间、导尿管留置时间、引流管留置时间、肛门恢复排气时间比较差异无统计学意义(P0.05);术后研究组性生活评分、生活质量评分高于对照组(P0.05),痛经程度评与对照组比较差异无统计学意义(P0.05);研究组压力性尿失禁、阴道顶端脱垂、直肠膨出发生率低于对照组(P0.05)。结论:改良腹腔镜手术治疗子宫腺肌症的治疗效果与传统腹腔镜手术相当,但患者性生活及生活质量较高,且对盆底支撑功能影响较小。     

Evaluation of the Rho A/Rho-kinase pathway in the uterus of the rat model of polycystic ovary syndrome

The aim of this study was to investigate the expression of RhoA/Rho-kinase in the uterus and the effect of Rho-kinase inhibitors on uterine contractions of dehydroepiandrosterone (DHEA) induced polycystic ovary syndrome (PCOS) rats. Forty-four female Sprague-Dawley (21 days old) rats divided into three groups: The control group (n?=?14, any procedure was not performed), vehicle group (n?=?14, 0.2?ml of sesame oil, subcutaneous injection, 20 days) and PCOS group (n?=?16, DHEA 6?mg/100?g in 0.2?ml of sesame oil, subcutaneous injection, 20 days). The myometrium thickness and uterine wet weight were assessed. The mRNA and protein expressions of Rho A, the effect of Rho-kinase inhibitors (fasudil and Y-27632) on KCl, carbachol, and PGF2α induced contractions were evaluated in the uterus. In the PCOS group, the myometrium thickness and uterine wet weight significantly increased compared to the control group and vehicle group. The mRNA expression level and the immunoreactive score of Rho A, ROCK 1, ROCK 2 were similar in all groups. In the PCOS group, KCl, carbachol, and PGF2α induced uterine contractions significantly increased compared to the control group and vehicle group. Fasudil and Y-27632 significantly inhibited KCl, carbachol, and PGF2α induced uterine contractions in all groups. In conclusion, the expression of Rho A, ROCK 1, ROCK 2 not changed although myometrium thickness, uterine wet weight and the contractile responses of uterus increased in the PCOS group. The results suggest that the Rho-kinase inhibitors effectively suppressed increased contractions in the PCOS group they might be potential therapeutic agents.