【作者】 郭艺红；

【导师】 孙莹璞； 乔玉环；

【作者基本信息】 郑州大学 ， 妇产科， 2002， 硕士

【摘要】 白细胞介素-1β（interleukin-1β，IL-1β）是一种重要的细胞因子和多肽调节因子，参与白细胞不同群体间信息传递，具有广泛的生物学活性，可以对多种免疫活性细胞具有调节作用。白血病抑制因子（leukaemia inhibitory factor，LIF）是一种高度糖机化的分泌型蛋白，具有促进骨髓白血病细胞系分化，抑制正常胚胎干细胞分化并保持其增殖能力等多种生物学活性。以往的研究认为IL-1β、LIF主要来源于血液中的单核细胞和巨噬细胞，最近的研究认为IL-1β、LIF还来源于卵泡液中的巨噬细胞、颗粒细胞和卵泡膜细胞等，且与生殖关系密切。迄今为止有关卵泡液中IL-1β及性激素水平与体外受精-胚胎移植（In vitro fertilization-embryo transfer,IVF-ET）结局关系的研究较少，尤其是多囊卵巢综合征（polycystic ovarian syndrome，PCOS）患者卵泡液中LIF及性激素水平与体外受精结局关系的研究甚少，国内尚未见报道。本研究采用酶联免疫双抗夹心法和时间分辨免疫荧光方法前瞻性研究了卵泡液中IL-1β、LIF的定量表达及雌二醇（Estradiol,E₂）和孕酮（Progesterone,P）的定量表达，旨在探2＊)2儿研卜研究’【嗽文 卵地液小IJ＊、1【，－】n及付激毁水。卜51VF刁Z”r纳而关系的研究讨卵巢局部几刁、LIF与性激素、IVFET结局之问的关系及规律，于价IL－ID、LIF在体外受龋－胚胎移植卜对口子受犯、胚胎发育及胚胎右床的作川和临床应用价俏。 研究对象H”OO年8厂－二001年6月在郑州人二孤一附属医院生殖中心行常规体外受精．胚胎移植不孕患者。第一部分：妊娠组23名不孕患者经IVFET后获得妊娠；非妊娠组 33名不孕患者经 IVFET后未获得妊娠。第二部分：11名 PCOS患者作为研究组，随机挑选 14名同期行 IVFET的不孕患者作为对照组。卵泡液为经阴道超声引导下抽吸排卵前卵泡所得。 研究方法1 采用酶联兔疫双抗夹心法检测卵泡液中LIF、IL刁 水平。2采用时间分辨免疫荧光方法检测卵泡液中 EZ和 P水平。3 体外受精书胎移植方法见我中心常规方法［’］。 结果 1．妊娠组与非奸娠组卵泡液IL二、LIF水平与性激索水平及IVF＊T结局的关系 1．1 卵泡液。卜1卜lp水平在妊娠组的浓度为 门3．62士12．18)p分ml，明显高 于非妊娠组门 ＊4土 15 ＊0)pyml中功刀5人而LIF浓度在妊娠组与非妊娠组的含量无明显差异（p＞0刀5）。 1．2 歹泡液＊ IL－ID水平和 I级胚胎形成率呈正相关 （＊0．352 P二0＊08）；卵泡液中IL－lfi和受精率亦呈正相关（r＝0．299 p—0＊25）。 2y)《)2儿帧＊J火’卜沦久 W州浓小门1。、川－1卜／扑卜淤术水丫‘J＊卜厂L叩入｛义九】扑叮叭 1．3 梦泡液叫。IL－lp和孕酮水平呈正 十关（，0．5”p、0＊01＼召泡液巳” 比值与受粕率成负相乡（FO．＊4P二0刀06）。 1．4 卵泡液＊ 水平与受粘率、I级胚胎形成率及孕酮水平无十关卜。 ZP＊OS组与对照组卵泡液几－lp、＊W 水平与性激素水平及IVFEI”结局的关系 二．IPCOS组卵泡液厂宇’LIF浓度为 ZI刀7pg／ml较对照组335沙g／lll明显降＜（＊＜O刀5） 而＊＊OS组L－ID的浓度为39．93pg／1lll I刃显高习对照组 28．30 pg／n。l（p＜0刀5）。 2．2 PCOS乡乒泡液中孕酮P浓度为 191．85 X 10’lllflol／L，对尸圭孕酮 P浓度为 305．89 XIO’nlnol／L，两者相比有明显差异…刃＊引；乒泡漱仁Q浓住＊＊＊S全为3334＊；l；n01／，叨显高于下照组＊兆互n。n＊／。（＊＜0刀5）。 二．3 ＊＊OS组和又只乡受粘率，二级胚刀形成率无明显差异（P＞0＊5）；＊＊OS 乡胚胎不植率为8．82％（3门4），对照组为20刀0％（9／45），PCOS组明 上 ｛ 于对照 组（P＜0刀5）；门床妊娠率 PCOS组为 18．18O（2／11），对二 组为 42．86o（／14），PCOS乡 明显低 刁又只乡（P＜0．05）。PCOS组OHSS发生率为27．27OO／11）明显高d对照组7．14o（／14）（P＜0．05）。 ＊4 卵泡液中LIF水平与E。水平在不孕组瞩COS组和对贝乡)呈负柏关（一0．442 p－0．027）：LIF水平与 LH／FSH比值在PCOS组呈负千Z关（r－0．682 p－0刀21），在不孕组（PCOS组不对照组）亦呈负个关（卜－O．至4p二O．OO75）。卵泡液中 32（）（）2年硕上研究生论文 卵泡液小 1．if。、l！．＊ fi及性傲紊水‘IL’jlVI。士丁拈n关系的研究IL－lp水平与 E。水平在 PCOS组呈正相关（r＝0＊12 p—0刀45）。在＊＊OS 组＊wF＊H＊值与P呈负相关（卜0．780 p二o＊05）。 2．SLIF与1卜lp水平两者门无相 关卜。 结论： l 好娠组与非妊娠组卵泡液几－［ p、LIF水平与性激素水平及IVF结局的关系研究表明： 1玉 口泡液卜。存刀I可F和 11J－lp。 1．2 卵泡液中的ILl 参与卵子受精过程，且和胚胎质量有关，可能是卵于受精和胚胎发育的营养因子。更多还原

【Abstract】 Interleukin-1 (IL-1 ) is an important cytokin and regulating factor of polypeptides, which takes part in message communication among different kinds of Leukocyte. It also has abroad spectrum of biological actions and functions of regulating various immunoactive cells. Leukaemia inhibitory factor (LIF)is a kind of highly glycosylated secretary protein .It has various biological functions of stimulating the differentiation of marrow leukaemia cell line, at the same time inhibits the normol embryo stem cells differentiation and matains the reproductive function . The previous researchers thought that IL-I , LIF mainly came from blood monocytes and macrophages ,but mordern studies indicate that IL-I , LIF are also released from macrophages , granulosa cells ,thecal ovarian cells ,ect. And have a close relationship with reproduction .Nevertheless, there has been few research on the relationships for IL-I ,sex hormone infollicuar fluid and the results of IVF -ET until today .There is no report about the relationships of LIF in follicuar fluid of the PCOS patients with sex hormone and result of IVF -ET in our country .The enzyme-linked immunosorbent assay(ELISA) is used to quantify IL-1 , LIF in follicuar fluid , and time-resolved fluoroimmunoassay is used to quantify estradiol and progesterone in follicular fluid.The purpose of our study was to investigate prospectively the relationships and rules of IL-I ,LIF , sex hormone in follicuar fluid and the results of IVF -ET , evaluate the action of IL-1 , LIF in insemination ,embryo development , implantation and assess their clinnical values . Patients: From August 2000 to June 200 1 , Patients undergoing treatment for IVF-ET were recruited from the reproductive center of The First Affiliated Hospital of Zheng Zhou University . Part I: Pregnancy group consisted of 23 infertile women who conceived after undergoing IVF -ET treatment. No pregnancy group consisted of 33 infertile women who failed to conceive after undergoing IVF -ET treatment. Part II: The study group consisted of 11 PCOS Patients,and the control group consisted of 14 infertile women. The follicular fluid was obstain on the day of oocyte retrieval.Methods: 1. The enzyme-linked immunosorbent assay(ELISA) is used to quantify IL-1 , LIF in follicuar fluid .2. The time-resolved fluoroimmunoassay is used toquantify estradiol and progesterone in follicular fluid.3 .The methods of I VF-ET reference to the routin ways in our center[1] Result1 The relationships of IL-I P , LIF, sex hormorne in follicle fluild and IVF-ET results in pregnancy group and non-pregnancy group1 . 1 The concentration of IL-1 3 in follicular fluid of pregnancy group is (33. 62 +12. 18) pg/ral, significantly higher than that in non-pregnancy group ( 25.74 + 15.00 ) pg/ml (p<0.05) .but there is no difference between the two group for the concentration of LIF in follicular fluid .1.2 The level of IL-1 P in follicular fluid is positively correlated with the grade I embryo rate ( r=0.352 P=0.008) ,and the fertilization rate (r-0.299 p=0.025) .1.3 The level of IL-1 & in follicular fluid is positively correlated with progesterone (r^O. 520 pO.OOl) .E2/P ratio is negatively con-elated with the fertilization rate(r=-0.364 p=0.006).1 .4 The level of LIF in follicular fluid is not correlated with the grade I embryo rate, the fertilization rate and Progesterone.2 The relationships of IL-I P , LIF, sex hormorne in follicle fluild and IVF-ET results in PCOS group and control group2.1 The concentration of LIF in follicular fluid of PCOS group(21.07pg/ml) is significantly lower than that in control group(33.53pg/ml) (p<0.05) . The concentration of IL-1 & in follicular fluid of PCOS group( 39.93pg/ml) is significantly higher than that in control group(28.30pg/ml) (p<0.05 ) .2.2 The concentration of Progesterone in follicular fluid of PCOS group(l 9 1.85X1 03nmol/L) is significantly lower than tha更多还原