【作者】 杨阳；

【导师】 郝桂敏； 曹金凤；

【作者基本信息】 河北医科大学 ， 妇产科学（专业学位）， 2015， 硕士

【摘要】 目的:本研究旨在通过观察多囊卵巢综合征(PCOS)患者子宫内膜上内源性大麻素受体CB1和水解酶FAAH的变化,比较其在月经周期不同时相与正常女性的差异,探讨内源性大麻素系统与PCOS的关系,旨在从分子水平更为深入的探究PCOS的发病机制,并为临床上该疾病的治疗提供新的思路。方法:选取2014.10-2015.1在河北医科大学附属第二医院生殖医学科门诊就诊的确诊为PCOS患者59例,平均年龄27±4岁。另外选择月经周期规律,排卵正常的女性患者54例为对照组,平均年龄28±5岁。两组患者均随机选择入组。研究对象均无心、肝、肾、甲状腺相关疾病,无子宫卵巢手术史,近三个月无激素类药物使用史,无输卵管积水等可能影响子宫内膜的疾病史。所有入组患者均在测定尿HCG阴性排除妊娠前提下行诊断性刮宫术,留取子宫内膜组织,PCOS组标本部分送病理及分期,刮出内膜组织经无菌纱布涂抹PBS冲洗去除多余血液,子宫内膜标本取体积小于0.5cm×0.5cm×0.1cm组织块4%四聚甲醛固定,乙醇脱水石蜡包埋,切片厚度3-5μm,每例标本做5张切片,用于免疫组织化学染色,图像经积分光密度分析,得出标本组织中CB1和FAAH含量。结果用SPSS13.0行统计学分析。结果:1免疫组化结果显示:CB1和FAAH主要表达在子宫内膜腺上皮的胞浆中,呈棕色颗粒状。2 PCOS组子宫内膜中CB1平均表达量在月经期、增殖期、分泌期分别为:334.85±130.63,345.03±87.91,382.94±117.82,各组平均表达量之间无明显统计学差异(P>0.05)。对照组子宫内膜中CB1平均表达量在月经期、增殖期、分泌期分别为:280.03±92.83,303.77±56.64,309.62±109.34,各组平均表达量之间无明显统计学差异(P>0.05)。3 PCOS组子宫内膜中FAAH平均表达量在月经期、增殖期、分泌期分别为:269.11±74.01,298.32±114.52,385.71±128.66,各组平均表达量之间有明显统计学差异(p<0.05),该差异在分泌期与月经期间有统计学意义(p<0.05),在分泌期与增殖期之间有统计学意义(p<0.05),而增殖期与月经期之间没有统计学意义(p>0.05)。对照组子宫内膜中faah平均表达量在月经期、增殖期、分泌期分别为:317.18±99.47,372.71±116.83,480.73±132.05,各组平均表达量之间差异有统计学意义(p<0.05),该差异在分泌期与月经期间有统计学意义(p<0.05),在分泌期与增殖期之间有统计学意义(p<0.05)。而增殖期与月经期之间没有明显统计学意义(p>0.05)。4月经期pcos组与对照组子宫内膜组织中cb1平均表达量为334.85±130.63、280.03±92.83,无明显统计学差异(p>0.05);增殖期pcos组与对照组阳性组织中cb1平均表达量为345.03±87.91、303.77±56.64,无明显统计学差异(p>0.05);分泌期pcos组与对照组阳性组织中cb1平均表达量为382.94±117.82、309.62±109.34,无明显统计学差异(p>0.05)。5月经期pcos组与对照组组织中faah平均表达量为269.11±74.01、317.18±99.47,无明显统计学差异(p>0.05);增殖期pcos组与对照组faah平均表达量为298.32±114.52、372.71±116.83,有明显统计学差异(p<0.05);分泌期pcos组与对照组faah平均表达量分别为385.71±128.66、480.73±132.05,有明显统计学差异(p<0.05)。结论:1正常女性子宫内膜中内源性大麻素受体cb1和水解酶faah主要表达在腺上皮的胞浆中。cb1在子宫内膜组织的表达与女性的月经周期时相无关。faah在子宫内膜组织的表达与女性月经周期时相有关,从增殖期到分泌期表达量有所升高,说明排卵后子宫内膜中内源性大麻素浓度处于低浓度水平,这可能有利于胚胎植入。2pcos患者子宫内膜组织中cb1受体的表达与正常女性无明显差异,cb1可能在pcos的发病和疾病演进的过程中未起到主要作用。而pcos患者子宫内膜组织中的faah平均表达量在增殖期和分泌期显著低于正常女性,说明高浓度的内源性大麻素水平可能是造成pcos患者非排卵性不孕、排卵后胚胎不易着床的原因之一。3内源性大麻素系统可能参与了PCOS疾病的发生与演进过程,该系统的调节与控制可能为治疗PCOS提供新的思路。更多还原

【Abstract】 Objective: To observe the expressions of the endogenous cannabinoid receptors CB1 and the hydrolytic enzymes FAAH in the endometrium at different menstrual phase in normal women and polycystic ovary syndrome patients, and compare the differences. Through this study, investigate the relationship between the endocannabinoid system and polycystic ovary syndrome,and explore the pathogenesis of polycystic ovary syndrome at molecular level. Try to provide a new clinical thought of the disease. Methods:Selected patients who were treated in the Reproductive Center of Second Hospital of Hebei Medical University from October 2014 to January 2015, including 59 cases of PCOS(experimental group),and 54 non-PCOS patients(control group).They were randomly selected. The average age was 27±4 and 28±5 respectively. All the patients had no heart, liver, kidney, thyroid-related diseases, had no ovary or uterus surgery, had no steroids drug use in the last three months, had no hydrosalpinx history which may affect the endometrium. All patients were negative urine HCG. All PCOS specimens were send to pathology test to determine stage.The Endometrial tissue was fixed in 4% paraformaldehyde, dehydrated by ethanol and embedded in paraffin. The slice thickness was 3-5um, 5 slices per specimen were immunohistochemistry stained and analysised by the integrated optical density, obtained tissue samples CB1 and FAAH content. Analysised the experimental data with SPSS software package. Results: 1 CB1 and FAAH mainly expressed in endometrial epithelial cytoplasm 2 The average expression of CB1 in PCOS’s endometrium during the menstrual period, the proliferative phase, the secretory phase were: statistically significant(P<0.05); in the secretory phase were 385.71±128.66、480.73±132.05, the difference is statistically significant(P <0.05). Conclusions: 1 female endometrial endogenous cannabinoid receptors CB1 and FAAH enzyme hydrolysis is expressed mainly in the cytoplasm of the glandular epithelium. CB1’s expression in the endometrial tissue has no relationship with women’s menstrual cycle. FAAH ’s expression in the endometrial tissue is related with the menstrual cycle phase, expression was increased in the amount from the proliferative phase to the secretion phase,indicating that within the endometrium after ovulation endocannabinoid concentrations are at low concentration levels, which may have conducive to embryo implantation. 2 PCOS endometrium CB1 receptor expression in normal women has no significant difference, indicating that CB1 may not play a major role in the onset and evolution of disease polycystic ovary syndrome. The average expression in patients with polycystic ovary syndrome, endometrial tissue volume FAAH in proliferative and secretory phase was significantly lower than normal women, indicating a high concentration of endogenous cannabinoid levels may be caused by non-ovulation in patients with polycystic ovary syndrome infertility, or it is another reason for difficult embryo implantation after ovulation. 3 endocannabinoid system may be involved in the pathogenesis of polycystic ovary syndrome and the evolution of the disease, regulation and control of the system may become a new target for the treatment of polycystic ovary syndrome.334.85±130.63, 345.03±87.91, 382.94±117.82, there is no significant difference between the average expression level of each group(P>0.05). The average expression of the non-PCOS’s CB1 during the menstrual period, the proliferative, and the secretory phase were: 280.03±92.83,303.77±56.64,309.62±109.34, there is no significant difference between the average expression level of each group(P>0.05). 3 The average expression of FAAH in PCOS’s endometrium during the menstrual period, the proliferative phase, secretory phase were: 269.11±74.01,298.32±114.52,385.71±128.66, the difference was statistically significant(P<0.05).The difference between the secretory phase and the menstrual period was statistically significant(P <0.05), between the secretory phase and the proliferative phase was statistically significant(P <0.05),between the proliferative phase and the menstrual period had no statistically significant(P>0.05).The average expression of non-PCOS’s FAAH during the menstrual period, the proliferative, and the secretory phase were: 317.18±99.47,372.71±116.83,480.73±132.05, the difference was statistically significant(P<0.05). The difference between the secretory phase and the menstrual period was statistically significant(P<0.05), between the secretory phase and the proliferative phase was statistically significant(P <0.05), between the proliferative phase and the menstrual period had no statistically significant(P> 0.05), 4 The average expressions of CB1-positive tissue of the PCOS group and the non-PCOS group in menstrual period endometrium were 334.85±130.63、280.03±92.83, no significant difference(P>0.05); in the proliferative phase were 355.10 ± 127.46, 268.60 ± 73.40, no significant difference(P>0.05); in the secretory phase were 382.94±117.82 、309.62±109.34, no significant difference(P>0.05). 5 The average expressions of FAAH-positive tissue of the PCOS group and the non-PCOS group in menstrual period endometrium were 269.11±74.01 、 317.18±99.47, no significant difference(P>0.05); in the proliferative phase were 298.32±114.52、372.71±116.83, the difference is更多还原