【作者】 刘秀；

【导师】 葛红山；

【作者基本信息】 大连医科大学 ， 妇产科学（专业学位）， 2023， 硕士

【摘要】 多囊卵巢综合征(polycystic ovary syndrome,PCOS)是常见的一种妇科疾病,具有的临床症状一般为排卵异常、月经紊乱、高雄激素和胰岛素抵抗等。全球多囊卵巢综合征的发病率在2%-26%。事实上,70%被诊断为多囊卵巢综合征的女性会继发不孕。越来越多的研究证明,PCOS与慢性炎症有关,炎症学说现已然成为大家研究的热点,但以往对PCOS慢性低度炎症的研究主要集中在血清或子宫内膜,而非卵泡液(follicular fluid,FF)。PCOS患者血清中多种炎症因子升高,例如TNF-α、IL-6、IL-8、IL-18等,而细胞因子被认为是卵泡微环境的主要成分,与感染和损伤引起的炎症一样,在促黄体生成素(luteotropic hormone,LH)激增或人绒毛膜促性腺激素(human chorionic gonadotropin,h CG)刺激下在卵巢中产生多种细胞因子,如IL-1、IL-6等。但在体外受精(in vitro fertilization,IVF)周期控制性超促排卵(controlled ovarian hyperstimulation,COH)过程中PCOS患者血清和卵泡液中不同时期下IL-8、IL-17A、IL-18的变化和表达仍需进一步研究。目的:探讨IVF周期中PCOS患者血清和卵泡液中IL-8、IL-17A和IL-18的表达和相关性分析,了解COH过程中卵泡早期血清、排卵前血清和卵泡液中炎症因子的变化,为炎症学说提供更多临床依据。材料与方法:(1)研究对象:本研究选取2021年11月-2023年1月就诊于泰州市人民医院生殖医学中心的进行IVF的PCOS患者98例和因其他因素导致不孕而就诊的对照组患者146例。(2)研究方法:查阅患者门诊医院HIS、LIS病历系统信息,收集患者年龄、身体质量指数、疾病史,以及行体外受精-胚胎移植术(in vitro fertilization-embryo transfer,IVF-ET)的术前常规检查包括血常规、空腹血糖、空腹胰岛素、血脂4项等结果。采用ELISA方法检测PCOS组和对照组患者卵泡早期血清、排卵前血清和卵泡液中IL-8、IL-17A和IL-18水平,比较三组样本中炎症因子的表达以及IVF周期中炎症因子的变化。结果:(1)PCOS组和对照组的BMI分别为24.69±4.31(kg/m~2)和22.62±3.64(kg/m~2);AMH分别为7.31±4.07和2.36±1.49;HOMA-IR分别为3.38±2.63和2.23±1.36;FIN分别为91.87±48.34(pmol/L)和66.82±37.28(pmol/L);T分别为1.82±1.00(mmol/L)和1.30±0.62(mmol/L);LDL分别为3.03±0.87(mmol/L)和2.78±0.64(mmol/L);WBC分别为6.62±1.88(10^9/L)和6.13±1.70(10^9/L);LH/FSH分别为0.87±0.74和0.66±0.53均高于对照组,差异均具有统计学意义。两组的年龄、FPG、TC、TG、VD均无明显差异。(2)PCOS患者卵泡液中IL-8的水平与FPG呈正相关(r=0.732,P<0.001),与TG同时呈正相关(r=0.518,P<0.05);IL-17A与糖脂代谢指标无明显相关;卵泡早期血清和排卵前血清中的IL-18水平与HDL-C呈明显负相关(r=-0.429,P<0.05;r=-0.562,P≤0.01)。(3)在对照组中,卵泡早期血清中IL-8、IL-17A和IL-18的浓度与排卵前血清浓度无明显差异(3.91±1.22pg/ml VS 3.89±1.98pg/ml,P>0.05;7.24±3.37pg/ml VS 7.06±4.12pg/ml,P>0.05;27.01±9.12pg/ml VS29.38±12.36pg/ml,P>0.05),卵泡早期和排卵前血清中IL-8、IL-17A和IL-18浓度小于卵泡液中浓度(3.91±1.22pg/ml、3.89±1.98pg/ml VS66.53±18.53pg/ml,P<0.001、P<0.001;7.24±3.37 pg/ml、7.06±4.12pg/ml VS 43.22±12.78 pg/ml,P<0.0001、P<0.001;27.01±9.12pg/ml、29.38±12.36pg/ml VS 34.85±9.53pg/ml,P<0.0001、P<0.05)。(4)在PCOS组中,卵泡早期血清中IL-8的浓度大于排卵前血清中IL-8浓度(4.07±1.25pg/ml VS 3.17±0.87pg/ml,P<0.01);卵泡早期血清中IL-17A的浓度小于排卵前血清中IL-17A浓度(7.41±3.11pg/ml VS17.09±14.37pg/ml,P<0.001);卵泡早期血清中IL-18的浓度与排卵前血清中IL-18浓度无明显差异(27.95±10.92pg/ml VS 26.96±8.82pg/ml,P>0.05)。其中卵泡早期血清和排卵前血清中IL-8、IL-17A和IL-18浓度均小于卵泡液中的浓度(4.07±1.25pg/ml、3.17±0.87pg/ml VS 76.86±31.12pg/ml,P<0.0001、P<0.001;7.41±3.11pg/ml、17.09±14.37pg/ml VS 55.52±12.34pg/ml,P<0.0001、P<0.001;27.95±10.92pg/ml、26.96±8.82pg/ml VS40.79±18.26pg/ml,P<0.0001、P<0.001;),差异均具有统计学意义。对照组和PCOS组相比,血清和卵泡液中IL-8和IL-18的浓度均无明显差异,对照组中排卵前血清和卵泡液中IL-17A的浓度均小于PCOS组,差异均具有统计学意义。结论:(1)PCOS组中BMI、AMH、HOMA-IR、FIN、LDL、LH/FSH、T、WBC明显高于对照组,说明PCOS女性存在肥胖、糖脂代谢与内分泌紊乱。(2)PCOS患者卵泡液中IL-8的水平与FPG和TG呈正相关,卵泡早期血清和排卵前血清中的IL-18水平与HDL-C呈负相关,其余未发现明显相关性,说明炎症因子与PCOS患者糖脂代谢紊乱有关。(3)PCOS人群排卵前血清和卵泡液中的IL-17A明显高于对照组人群,说明PCOS人群在排卵过程中的炎症程度高于对照组人群。(4)不论在PCOS人群还是对照组人群,卵泡液中IL-8、IL-17A和IL-18的水平明显高于血清中水平,说明排卵过程中产生的炎症因子主要在卵泡液中产生并积聚。更多还原

【Abstract】 polycystic ovary syndrome(PCOS)is a common gynecological disease,mainly characterized by abnormal ovulation,menstrual disorder,hyperandrogenemia and insulin resistance.The global incidence of polycystic ovary syndrome ranges from 2% to 26%.In fact,70% of women diagnosed with PCOS go on to have secondary infertility.More and more studies have proved that PCOS is related to chronic inflammation,and the inflammation theory has become a hot research topic.However,previous studies on chronic low inflammation of PCOS mainly focused on serum or endometrial,rather than follicular fluid(FF).Studies have shown that the serum levels of various inflammatory factors in PCOS patients increase,such as TNF-α,IL-6,IL-8,IL-18,etc.In addition,cytokines are considered to be the main components of the follicular microenvironment.As with inflammation caused by infection and injury,it produce cytokines in the ovaries when stimulated by a surge of luteotropic hormone(LH)or human chorionic gonadotropin(h CG),Such as IL-1,IL-6 and so on.But in vitro fertilization(IVF)cycle controlled ovarian hyperstimulation,the changes and expressions of IL-8,IL-17A and IL-18 in serum and follicular fluid of PCOS patients at different periods during COH process still need further study.Objective: To investigate the expression and correlation analysis of IL-8,IL-17A and IL-18 in serum and follicular fluid of PCOS patients during IVF cycle,and to understand the changes of inflammatory factors in serum at the early stage of follicle,serum before ovulation and follicular fluid during COH process,so as to provide more clinical basis for inflammation theory.Materials and Methods:(1)Subjects: 98 patients with PCOS who underwent in vitro fertilization-embryo transfer(IVF-ET)and 146 patients in the control group who were treated for infertility due to other factors were selected in this study from November 2021 to January 2023 in the Reproductive Medicine Center of Taizhou People’s Hospital.(2)Research methods: The information of HIS and LIS system in the outpatient department of the patient was consulted,and the patient’s age,weight and disease history were collected,as well as the preoperative routine examinations including blood routine,fasting blood glucose,fasting insulin and blood lipid,etc.The levels of IL-8,IL-17A and IL-18 in early follicular serum,pre-ovulation serum and follicular fluid of PCOS group and control group were detected by ELISA,and the expression of inflammatory factors in the three groups and the changes of inflammatory factors during IVF cycle were compared.Results:(1)The BMI of PCOS group and control group was 24.69±4.31(kg/m2)and22.62±3.64(kg/m2),respectively.AMH were 7.31±4.07 and 2.36±1.49,respectively.HOMA-IR were 3.38±2.63 and 2.23±1.36,respectively;FIN were 91.87±48.34(pmol/L)and 66.82±37.28(pmol/L),respectively.T was 1.82±1.00(mmol/L)and1.30±0.62(mmol/L),respectively.LDL was 3.03±0.87(mmol/L)and 2.78±0.64(mmol/L),respectively.WBC were 6.62±1.88(10^9/L)and 6.13±1.70(10^9/L),respectively.The LH/FSH values of 0.87±0.74 and 0.66±0.53 were higher than those of the control group,and the differences were statistically significant.There were no significant differences in age,FPG,TC,TG and VD between the two groups.(2)The level of IL-8 in follicular fluid of PCOS patients was positively correlated with FPG(r=0.732,P<0.001)and TG(r=0.518,P<0.05).There was no significant correlation between IL-17A and glycolipid metabolism indexes.The levels of IL-18 in serum at the early stage of follicle and serum before ovulation were negatively correlated with HDL-C(r=-0.429,P<0.05;r=-0.562,P≤0.01).(3)In the control group,there was no significant difference between the concentrations of IL-8,IL-17A and IL-18 in the serum in the early follicle period and the serum concentrations before ovulation(3.91±1.22pg/ml VS 3.89±1.98pg/ml,P>0.05;7.24±3.37pg/ml VS 7.06±4.12pg/ml,P>0.05;27.01±9.12pg/ml VS29.38±12.36pg/ml,P>0.05),IL-8,IL-17A and IL-18 concentrations in early follicle and preovulation serum were less than those in follicular fluid(3.91±1.22pg/ml,3.89±1.98pg/ml VS 66.53±18.53pg/ml,P<0.001,P< 0.001;7.24±3.37 pg/ml 、7.06±4.12pg/ml VS 43.22±12.78 pg/ml,P<0.0001、P<0.001;27.01±9.12pg/ml、29.38±12.36pg/ml VS 34.85±9.53pg/ml,P<0.0001 、P<0.05).(4)In the PCOS group,the concentration of IL-8 in the serum in the early follicle period was greater than the concentration of IL-8 in the serum before ovulation(4.07±1.25pg/ml VS 3.17±0.87pg/ml,P<0.01);The concentration of IL-17A in the serum of early follicularity is less than that in the serum before ovulation(7.41±3.11pg/ml VS 17.09±14.37 pg/ml,P<0.001);There was no significant difference between the concentration of IL-18 in the serum of early follicles and the concentration of IL-18 in preovulation serum(27.95±10.92 pg/ml VS 26.96±8.82pg/ml,P>0.05).Among them,the concentrations of IL-8,IL-17A and IL-18 in early follicular serum and preovulation serum were smaller than those in follicular fluid(4.07±1.25pg/ml 、 3.17±0.87pg/ml VS 76.86±31.12pg/ml,P<0.0001,P<0.001;7.41±3.11pg/ml、17.09±14.37pg/ml VS 55.52±12.34 pg/ml,P<0.0001、P<0.001;27.95±10.92pg/ml 、 26.96±8.82pg/ml VS 40.79±18.26pg/ml,P<0.0001 、P<0.001;),the differences were statistically significant.Compared with the PCOS group,there was no significant difference in the concentrations of IL-8 and IL-18 in serum and follicular fluid,and the concentrations of IL-17A in preovulation serum and follicular fluid in the control group were lower than those in the PCOS group,and the differences were statistically significant.Conclusion:(1)BMI,AMH,HOMA-IR,FIN,LDL,LH/FSH,T,WBC in the PCOS group were significantly higher than those in the control group,indicating that women with PCOS had obesity,glycolipid metabolism and endocrine disorders.(2)The level of IL-8 in follicular fluid in PCOS patients was positively correlated with FPG and TG,and the level of IL-18 in early follicular serum and preovulation serum was negatively correlated with HDL-C,and no significant correlation was found in the rest,indicating that inflammatory factors can cause disorders of glycolipid metabolism in PCOS patients.(3)The IL-17A in the preovulation serum and follicular fluid in the PCOS population was significantly higher than that in the control group,indicating that the degree of inflammation in the PCOS population during ovulation was higher than that in the control group.(4)Whether in the PCOS population or the control group,the levels of IL-8,IL-17A and IL-18 in the follicular fluid were significantly higher than the serum levels,indicating that the inflammatory factors produced during ovulation were mainly produced and accumulated in the follicular fluid.更多还原