本文選題：PAF + 卵泡液��； 參考：《河北醫(yī)科大學(xué)》2014年碩士論文

【摘要】：研究目的：通過體外受精-胚胎移植(In Vitro Fertilization embryoTransfer, IVF-ET)助孕的患者卵泡液中血小板激活因子(Platelet ActivatingFactor, PAF)水平及其與年齡、體重指數(shù)(Body mass index, BMI)、雙側(cè)卵巢竇卵泡數(shù)(Antral follicle count, AFC)、性激素及實(shí)驗(yàn)室參數(shù)的關(guān)系研究，以此來評(píng)價(jià)PAF在卵子發(fā)育、受精、胚胎發(fā)育中的作用及對(duì)妊娠結(jié)局的影響，為尋找一種能夠客觀、準(zhǔn)確評(píng)價(jià)卵子質(zhì)量及胚胎發(fā)育潛能的新方法提供理論依據(jù)。 材料和方法： 選擇2013年7月1日~2013年8月31日于河北醫(yī)科大學(xué)第四醫(yī)院生殖中心首次接受IVF-ET（包括常規(guī)IVF和卵泡漿內(nèi)單精子注射Intracytoplasmicsperm injection, ICSI）并在本周期移植患者共162例，作為研究對(duì)象，并排除女性既往患有結(jié)核、子宮內(nèi)膜異位癥及宮腔病變，近3個(gè)月未應(yīng)用過激素類藥物，既往無自然流產(chǎn)史，夫妻雙方染色體正常，未發(fā)生卵巢過度刺激綜合征（Ovarian hyperstimulation syndrome, OHSS）等。 根據(jù)患者的情況選擇標(biāo)準(zhǔn)長(zhǎng)方案或微刺激方案進(jìn)行控制性卵巢刺激。根據(jù)患者采集的精液情況選擇常規(guī)IVF或ICSI方法授精。取卵后48h、72h觀察胚胎發(fā)育情況。優(yōu)質(zhì)胚胎標(biāo)準(zhǔn)：取卵后72h胚胎細(xì)胞數(shù)≥7且≤12個(gè)，卵裂球大小均勻或略不均勻，碎片≤20%的胚胎為優(yōu)質(zhì)胚胎。 為了研究卵泡液PAF水平與年齡、BMI、基礎(chǔ)卵泡刺激素（Basalfollicle-Stimulating Hormone, bFSH）、基礎(chǔ)卵泡刺激素/基礎(chǔ)黃體生成素（Basal luteinizing hormone, bLH）、AFC、注射絨毛膜促性腺激素(Humanchorionic gonadotropin, HCG)日血清雌二醇(Estradiol, E2)和孕酮(Progesterone, P)、獲卵數(shù)的關(guān)系，將162例患者以35歲為臨界，分析年齡≤35歲和年齡＞35歲患者卵泡液中PAF水平的差異；以BMI=25kg/m2為臨界，分析BMI＜25kg/m2和BMI≥25kg/m2患者卵泡液中PAF水平的差異；以bFSH=10IU/L為臨界，分析bFSH≤10IU/L和bFSH＞10IU/L患者卵泡液中PAF水平的差異；以bFSH/bLH=2為臨界，分析bFSH/bLH≤2和bFSH/bLH＞2患者卵泡液中PAF水平的差異；以AFC=7個(gè)為臨界，分析AFC≤7個(gè)和AFC＞7個(gè)患者卵泡液中PAF水平的差異；以HCG日血清E2=1000pg/ml為臨界，分析HCG日血清E2≤1000pg/ml和HCG日血清E2＞1000pg/ml患者卵泡液中PAF水平的差異；以HCG日血清P=2.6ng/ml為臨界，分析HCG日血清P≤2.6ng/ml和HCG日血清P＞2.6ng/ml患者卵泡液中PAF水平的差異；以獲卵數(shù)=4個(gè)為臨界，分析獲卵數(shù)≤4個(gè)和獲卵數(shù)＞4個(gè)患者卵泡液中PAF水平的差異。 為了進(jìn)一步研究卵泡液中PAF水平對(duì)相同條件人群卵子質(zhì)量及胚胎發(fā)育的影響，將同時(shí)滿足非多囊卵巢綜合癥（Polycystic ovary syndrome,PCOS)、年齡≤35歲、BMI＜25kg/m2、AFC＞7個(gè)及應(yīng)用標(biāo)準(zhǔn)長(zhǎng)方案的68例移植患者，按卵泡液中PAF水平分為三組：①P AF≤5.9ng/ml組（17例），②5.9＜PAF＜11ng/ml組（34例），③P AF≥11ng/ml組（17例），在三組的年齡、不孕年限、BMI等基礎(chǔ)情況及Gn天數(shù)、Gn用量、獲卵數(shù)及移植日子宮內(nèi)膜厚度等COS情況均處于同一水平的前提下比較三組患者受精率、雙原核(Pronuclear,2PN)形成率、優(yōu)胚率、植入率及妊娠率的差異。 應(yīng)用電化學(xué)發(fā)光法(Electro-Chemiluminescence Immuno assay, ECLIA)檢測(cè)上述患者血清中的E2、P、LH、FSH及HCG水平；應(yīng)用酶聯(lián)免疫吸附法(Enzyme-linked immunosorbent assay, ELISA)檢測(cè)患者取卵日第一個(gè)穿刺的成熟卵泡液的PAF水平。 統(tǒng)計(jì)學(xué)處理：數(shù)據(jù)統(tǒng)計(jì)分析采用SPSS13.0版軟件包，計(jì)量資料結(jié)果以均數(shù)±標(biāo)準(zhǔn)差(x±s)表示，比較采用t檢驗(yàn)、方差分析、秩和檢驗(yàn)、x2檢驗(yàn)，相關(guān)性分析采用pearson及spearman相關(guān)分析。P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果: 1年齡＞35歲患者卵泡液中PAF水平明顯低于年齡≤35歲患者（P＜0.05）。 2BMI＜25kg/m2和BMI≥25kg/m2患者之間的卵泡液中PAF水平，無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 3bFSH≤10IU/L和bFSH＞10IU/L患者之間的卵泡液PAF水平無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 4bFSH/bLH≤2患者卵泡液中PAF水平高于bFSH/bLH＞2患者，但無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 5AFC≤7個(gè)和AFC＞7個(gè)患者卵泡液中PAF水平無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 6HCG日血清E2水平＞1000pg/ml患者卵泡液PAF水平高于HCG日血清E2≤1000pg/ml，但無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 7HCG日血清P≤2.6ng/ml和HCG日血清P＞2.6ng/ml患者卵泡液中PAF水平無統(tǒng)計(jì)學(xué)差異(P＞0.05)。 8獲卵數(shù)≤4個(gè)患者卵泡液中PAF水平低于獲卵數(shù)＞4個(gè)患者，但無統(tǒng)計(jì)學(xué)差異(P＞0.05)。 9卵泡液中PAF水平與年齡、BMI、bFSH、bFSH/bLH、AFC、HCG日血清E2和P水平、獲卵數(shù)的相關(guān)性分析：卵泡液中PAF水平與年齡呈明顯負(fù)相關(guān)（r=-0.197，P＜0.05）；PAF與BMI(r=0.006，P＞0.05)、bFSH(r=0.097，P＞0.05)、bFSH/bLH(r=0.039，P＞0.05)、AFC(r=0.098，P＞0.05)、HCG日血清E2水平(r=-0.101，P＞0.05)、HCG日血清P水平(r=0.066，P＞0.05)、獲卵數(shù)(r=0.015，P＞0.05)無明顯相關(guān)性，但發(fā)現(xiàn)HCG日血清E2以1000pg/ml為界，PAF與之有不同相關(guān)性，當(dāng)HCG日血清E2≤1000pg/ml時(shí)，卵泡液中PAF水平與之呈明顯正相關(guān)（r=0.567，P＜0.05）；而HCG日血清E2水平＞1000pg/ml時(shí)，卵泡液中PAF與之呈明顯負(fù)相關(guān)（r=-0.242，P＜0.05）。 10年齡、不孕年限、BMI等基礎(chǔ)情況及應(yīng)用促性腺激（Gonadotrophin,Gn)天數(shù)、Gn用量、獲卵數(shù)及移植日子宮內(nèi)膜厚度等COS情況均無統(tǒng)計(jì)學(xué)差異的68例移植患者按卵泡液中PAF水平分為PAF≤5.9ng/ml、5.9＜PAF＜11ng/ml和PAF≥11ng/ml三組。三組的受精率和植入率隨著PAF水平的升高而升高，但無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 11三組的2PN形成率有統(tǒng)計(jì)學(xué)差異，PAF≥11ng/ml組患者的2PN形成率在三組中最高，為71.16%（P＜0.05），且明顯高于PAF≤5.9ng/ml組的59.81%，兩組比較有統(tǒng)計(jì)學(xué)差異（P＜0.05），而PAF≥11ng/ml組和5.9＜PAF＜11ng/ml組、PAF≤5.9ng/ml組和5.9＜PAF＜11ng/ml組比較無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 12三組的優(yōu)胚率有統(tǒng)計(jì)學(xué)差異，PAF≥11ng/ml組患者的優(yōu)胚率在三組中最高，為64.96%（P＜0.05），明顯高于PAF≤5.9ng/ml組的46.09%，兩組比較有統(tǒng)計(jì)學(xué)差異（P＜0.05），也明顯高于5.9＜PAF＜11ng/ml組的50.63%，兩組比較有統(tǒng)計(jì)學(xué)差異（P＜0.05），但PAF≤5.9ng/ml組和5.9＜PAF＜11ng/ml組比較無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 13三組的妊娠率有統(tǒng)計(jì)學(xué)差異，PAF≥11ng/ml組患者的妊娠率在三組中最高，為70.59%（P＜0.05），且明顯高于PAF≤5.9ng/ml組的23.53%，兩組比較有統(tǒng)計(jì)學(xué)差異（P＜0.05），而PAF≥11ng/ml組和5.9＜PAF＜11ng/ml組、PAF≤5.9ng/ml組和5.9＜PAF＜11ng/ml組比較無統(tǒng)計(jì)學(xué)差異（P＞0.05）。 結(jié)論：PAF是可以促進(jìn)卵子發(fā)育的細(xì)胞因子，卵泡液中PAF水平受年齡影響。卵巢功能正常的患者取卵日卵泡液PAF水平較高。HCG日血清不同的E2水平對(duì)卵泡液PAF水平有不同影響，PAF不僅可以促進(jìn)顆粒細(xì)胞E2的分泌，E2過高會(huì)導(dǎo)致PAF水平的下降，在IVF助孕過程中，，要適度控制COS程度，減少過高E2產(chǎn)生及獲卵數(shù)的過多，從而改善IVF結(jié)局。取卵日卵泡液PAF水平可以作為預(yù)測(cè)妊娠結(jié)局的指標(biāo)，也可以作為一種評(píng)價(jià)卵子質(zhì)量及胚胎發(fā)育潛能的新方法。
[Abstract]:Objective: the level of platelet activating factor (Platelet ActivatingFactor, PAF) in follicular fluid of patients with In Vitro Fertilization embryoTransfer (IVF-ET) and its age, body mass index (Body mass index, BMI), double ovarian follicles and ovarian follicles, sex hormones and reality In order to evaluate the effect of PAF on the development of egg, fertilization, embryo development and the effect on the outcome of pregnancy, this study provides a theoretical basis for finding a new method that can objectively and accurately evaluate the quality of egg and the potential of embryonic development.
Materials and methods:
For the first time in the reproductive center of the fourth hospital of Hebei Medical University, July 1, 2013 ~2013, IVF-ET (including Intracytoplasmicsperm injection, ICSI) in the routine IVF and follicle syrup were given for the first time, and 162 cases were transplanted in this period. In the last 3 months, no hormone drugs have been used. There was no history of spontaneous abortion. The chromosomes of both husband and wife were normal, and no ovarian hyperstimulation syndrome (Ovarian hyperstimulation syndrome, OHSS) occurred.
According to the patient's condition, the control ovarian stimulation was selected by the standard long prescription or microstimulus. According to the condition of the semen collected by the patient, the conventional IVF or ICSI method was selected. The embryo development was observed by 48h and 72h after the egg. The standard of high quality embryos: the number of 72h embryos after the oocyte was more than 7 and less than 12, and the size of the blastomere was uniform or slightly uneven. The embryo of less than 20% of the fragment is a good embryo.
In order to study the PAF level and age of follicular fluid, BMI, basal follicle stimulating hormone (Basalfollicle-Stimulating Hormone, bFSH), basal follicle stimulating hormone / basal luteinizing hormone (Basal luteinizing hormone, bLH), AFC, and serum estradiol (Humanchorionic gonadotropin) and progesterone (Humanchorionic gonadotropin). Sterone, P), the relationship between the number of eggs was obtained. The difference in the level of PAF in follicular fluid of age < 35 years and age > 35 years old was analyzed in 162 patients at the age of 35 years. The difference of PAF levels in the follicle fluid of BMI < 25kg/m2 and BMI > 25kg/m2 patients was analyzed with BMI=25kg/m2 as critical. The difference in the level of PAF in the follicular fluid of the patients and the difference in the level of PAF in the follicular fluid of bFSH/bLH < 2 and bFSH/bLH > 2 were analyzed with bFSH/bLH=2 as critical. The difference of PAF levels in the follicular fluid of AFC less than 7 and AFC > 7 were analyzed with AFC=7 as critical. The difference in the level of PAF in follicular fluid of patients with E2 > 1000pg/ml in daily serum; the difference between HCG day serum P < 2.6ng/ml and HCG day serum P > 2.6ng/ml patient's PAF level was analyzed with HCG day serum P=2.6ng/ml, and the difference between the number of ovum number less than 4 and the number of ovum more than 4 patients was analyzed.
In order to further study the effect of PAF level in follicular fluid on oocyte quality and embryo development in the same condition population, it will meet non polycystic ovarian syndrome (Polycystic ovary syndrome, PCOS), age less than 35 years old, BMI < 25kg/m2, AFC > 7 and 68 cases of applied standard long prescription, divided into three groups according to the level of PAF in the follicular fluid: (1) P AF < 5.9ng/ml group (17 cases), 5.9 < PAF < 11ng/ml group (34 cases), P AF > 11ng/ml group (17 cases). In the three groups of age, infertile years, BMI and other basic conditions and Gn days, Gn dosage, the number of eggs and the thickness of the endometrium of the transplant day were at the same level and compared the fertilization rate of the three groups. The difference in the rate of adult, the rate of optimal embryo, the rate of implantation and the rate of pregnancy.
Electro-Chemiluminescence Immuno assay (ECLIA) was used to detect the levels of E2, P, LH, FSH and HCG in the serum of the above patients. The level of the mature follicle fluid was detected by enzyme linked immunosorbent assay (Enzyme-linked immunosorbent assay).
Statistical analysis: SPSS13.0 software package was used for statistical analysis of data, and the results of measurement data were expressed with mean standard deviation (x + s), and t test, variance analysis, rank sum test, x2 test, and correlation analysis with Pearson and Spearman correlation analysis.P < 0.05 were statistically significant.
Result:
1 the level of PAF in follicular fluid of patients aged 35 to 35 was significantly lower than that of patients younger than 35 years old (P < 0.05).
PAF levels in follicular fluid between patients with 2BMI < 25kg/m2 and BMI > 25kg/m2 were not significantly different (P > 0.05).
There was no significant difference in PAF level of follicular fluid between patients with 3bFSH < 10IU/L and bFSH > 10IU/L (P > 0.05).
The level of PAF in follicular fluid of patients with 4bFSH/bLH < 2 was higher than that of bFSH/bLH > 2, but there was no significant difference (P > 0.05).
PAF levels in follicular fluid of patients with 5AFC < 7 and AFC > 7 patients were not significantly different (P > 0.05).
The level of serum E2 in patients with 6HCG > 1000pg/ml > PAF level was higher than that in HCG day serum E2 < 1000pg/ml, but there was no significant difference (P > 0.05).
There was no significant difference in PAF level in follicular fluid of patients with serum P < 2.6ng/ml and HCG days in serum 7HCG > 2.6ng/ml on day 7HCG (P > 0.05).
8 the level of PAF in follicular fluid was lower than that in 4 patients, but there was no significant difference between the 4 patients (P > 0.05).
9 the level of PAF in follicular fluid and age, BMI, bFSH, bFSH/bLH, AFC, HCG day levels of serum E2 and P, and the correlation analysis of the number of eggs obtained: the level of PAF in the follicle fluid was negatively correlated with age (r=-0.197, P < 0.05); (r=-0.101, P > 0.05), the level of serum P (r=0.066, P > 0.05) on HCG day, the number of acquired eggs (r=0.015, P > 0.05) was not significantly correlated, but it was found that HCG day serum E2 was bounded by 1000pg/ml, and there was a different correlation between PAF and it. 1000pg/ml showed a negative correlation with PAF in follicular fluid (r=-0.242, P < 0.05).
The 10 age, infertile years, BMI and other basic conditions as well as the use of gonadotropin (Gonadotrophin, Gn) days, the amount of Gn, the number of eggs and the thickness of the endometrium in the transplantation day were not statistically different in the 68 cases of transplant patients, which were divided into PAF < 5.9ng/ml, 5.9 < PAF < 11ng/ml and PAF > 11ng/ml three. Three groups of fertilization rate and planting. The intake rate increased with the increase of PAF level, but there was no significant difference (P > 0.05).
11 the formation rate of 2PN in the three groups was statistically different. The 2PN formation rate in the group of PAF > 11ng/ml was the highest in the three groups, 71.16% (P < 0.05), and significantly higher than that in the PAF < 5.9ng/ml group. The two groups were statistically different (P < 0.05), and PAF > 11ng/ml group and 5.9 < PAF < 11ng/ml group. 5.9 Statistical difference (P > 0.05).
12 the optimal embryo rate in the three groups was statistically different. The best embryo rate in the PAF group was the highest in the three groups, 64.96% (P < 0.05), which was significantly higher than the 46.09% in the PAF < 5.9ng/ml group. The two groups were statistically different (P < 0.05), and were significantly higher than 50.63% in the 5.9 < PAF < 11ng/ml group. There was a statistically significant difference between the group two (P < 0.05), but PAF < < 5.9ng/ml. There was no significant difference between the group and 5.9 < PAF < 11ng/ml group (P > 0.05).
13 the pregnancy rate in the three groups was statistically different. The pregnancy rate in the group PAF > 11ng/ml was the highest in the three groups, 70.59% (P < 0.05), and significantly higher than that in the PAF group (23.53%). The two groups were statistically different (P < 0.05), while PAF > 11ng/ml and 5.9 PAF < 11ng/ml, PAF < 5.9ng/ml group and 5.9 < 5.9 Differences in learning (P > 0.05).
Conclusion: PAF is a cytokine that can promote the development of egg. The level of PAF in follicular fluid is influenced by age. The level of PAF in the follicle fluid of the ovarian follicle fluid is higher on.HCG day and the level of E2 has different effects on the level of PAF in follicular fluid. PAF can not only promote the secretion of granular E2, but the high E2 will lead to the decrease of PAF level. In the process of IVF pregnancy, it is necessary to control the degree of COS moderately, reduce the excessive E2 production and the excessive number of eggs, thus improve the IVF outcome. The PAF level of the egg day follicle can be used as an index for predicting the outcome of pregnancy, and can also be used as a new method to evaluate the quality of egg and the potential of embryonic development.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R714.8

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