重组人卵泡刺激素作用下卵泡黄素化颗粒细胞M-CSF及受体表达的临床差异

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (12): 1415-1420.

重组人卵泡刺激素作用下卵泡黄素化颗粒细胞M-CSF及受体表达的临床差异

余蓉，金昊，林金菊，黄学锋

温州医科大学附属第一医院生殖医学中心，温州 325000，浙江

收稿日期:2017-06-09 修回日期:2017-07-10 出版日期:2017-12-26 发布日期:2018-01-02
作者简介:余蓉，女，硕士研究生，主治医师，研究方向：生殖内分泌。 Tel:13615779108 E-mail:nerve1116@163.com
基金资助:
温州市科技局项目（Y20130375）

Comparison of M-CSF and its receptor mRNA expression in luteinized granulosa cells after using recombinant human follicle stimulating hormone

YU Rong, JIN Hao, LIN Jinju, HUANG Xuefeng

Reproductive Medical Center, the First Affiliate Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, China

Received:2017-06-09 Revised:2017-07-10 Online:2017-12-26 Published:2018-01-02

摘要/Abstract

摘要：

目的:比较使用重组人卵泡刺激素（r-FSH）后，患者卵泡黄素化颗粒细胞巨噬细胞集落刺激因子（M-CSF）及其受体（M-CSFR）mRNA表达的差异，以探讨M-CSF及其受体对卵巢反应性的影响。方法: 接受体外受精治疗的96例多囊卵巢综合征（PCOS）患者和157例非PCOS患者，根据其使用r-FSH后反应性不同分为四组：即PCOS快、慢反应组和非PCOS快、慢反应组。收集成熟卵泡穿刺后黄素化颗粒细胞，采用SYBR Green荧光定量RT-PCR法检测颗粒细胞M-CSF、M-CSFR和管家基因GAPDH的mRNA表达，通过比较阈值法（CT值目的基因－CT值管家基因）进行基因相对定量。双独立样本t检验比较组间基因表达差异，并对差异有统计学意义的指标行Spearman相关分析检验。结果:各组患者r-FSH使用量无统计学差异（P>0.05）。PCOS患者整体与非PCOS患者整体比较，M-CSF和M-CSFR的mRNA定量均无统计学差异（P>0.05）。分组比较后，各组间M-CSF mRNA定量无统计学差异（P>0.05）。而PCOS慢反应组M-CSFR mRNA定量低于快反应组患者，差异有统计学意义（P=0.006）。对PCOS组颗粒细胞M-CSFR mRNA定量和r-FSH使用天数的Spearman相关分析提示，两者相关性有统计学意义（P=0.023），相关系数0.511。结论:PCOS卵巢慢反应患者对促排卵药物反应迟缓可能与其颗粒细胞M-CSFR表达减少有关。M-CSF及其受体参与影响卵巢对r-FSH的反应性。

关键词: 巨噬细胞集落刺激因子, 卵泡刺激素, 多囊卵巢综合征, 黄素化颗粒细胞

Abstract:

AIM: To explore the effect of M-CSF and its receptor on the response of ovarian stimulation by comparing the expression of macrophage colony stimulating factor (M-CSF) and its receptor mRNA in luteinized granulosa cells in patients after using combinant human follicle stimulating hormone. METHODS: Ninety-six patients with polycystic ovary syndrome (PCOS) and 157 patients with non-PCOS underwent in vitro fertilization were divided into four groups, i.e. the PCOS fast and slow reaction group, and the non PCOS fast and slow reaction group, according to their response to recombinant human follicle stimulating hormone (r-FSH). Luteinized granulosa cells were then collected after mature follicular puncture. SYBR Green quantitative RT-PCR method was used to detect the expression of M-CSF, M-CSFR and GAPDH in the mRNA gene of the granulosa cells samples. The relative quantity of these genes were determined by comparing the threshold value (CT value of the target gene subtract CT value of housekeeping gene). The difference of gene expression between two groups was compared by t test, and Spearman correlation analysis was used to describe the data relationships. RESULTS: No significant difference was observed in the use of r-FSH among the different groups (P>0.05). Neither was there any significant difference in mRNA quantity of M-CSF or M-CSFR between the entire PCOS and non PCOS patients (P>0.05). After grouping, no significant difference was observed between any two groups in the expression of M-CSF(P>0.05). The expression of M-CSFR in PCOS slow response group was significantly lower than that of PCOS fast response group (P=0.006). Meanwhile, the Spearman analysis showed that the correlation between the quantification of M-CSFR mRNA and the days of r-FSH in PCOS group was statistically significant(P=0.023); the correlation coefficient was 0.511. CONCLUSION: The slow response to ovarian stimulation in PCOS patients is possibly related to the reduction of granulocyte M-CSFR expression. The M-CSF and its receptor may be involved in the ovarian stimulation response process.

Key words: macrophage colony stimulatory factor, human follicle stimulating hormone, polycystic ovary syndrome, luteinized granulosa cells

中图分类号:

R969

余蓉，金昊，林金菊，黄学锋. 重组人卵泡刺激素作用下卵泡黄素化颗粒细胞M-CSF及受体表达的临床差异[J]. 中国临床药理学与治疗学, 2017, 22(12): 1415-1420.

YU Rong, JIN Hao, LIN Jinju, HUANG Xuefeng . Comparison of M-CSF and its receptor mRNA expression in luteinized granulosa cells after using recombinant human follicle stimulating hormone[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(12): 1415-1420.

参考文献

［1］Zhou W, Chu D, Sha W, et al. Effects of granulocyte-macrophage colony-stimulating factor supplementation in culture medium on embryo quality and pregnancy outcome of women aged over 35 years［J］. J Assist Reprod Genet, 2016, 33(1): 39-47.
［2］Ziebe S, Loft A, Povlsen BB, et al. A randomized clinical trial to evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium for in vitro fertilization［J］. Fertil Steril, 2013, 99(6): 1600-1609.
［3］Cohen J, Chabbert-Buffet N, Darai E. Diminished ovarian reserve, premature ovarian failure, poor ovarian responder--a plea for universal definitions［J］. J Assist Reprod Genet, 2015, 32(12): 1709-1712.
［4］Handelsman DJ, Teede HJ, Desai R, et al. Performance of mass spectrometry steroid profiling for diagnosis of polycystic ovary syndrome［J］. Hum Reprod, 2017, 32(2): 418-422.
［5］吴小华, 程立立, 张敏, 等. 卵母细胞分泌因子在人卵丘颗粒细胞中的表达及其与卵母细胞发育潜能的关系［J］. 生殖与避孕, 2016, 36(9): 25-28.
［6］Salmassi A1, Zhang Z, Schmutzler AG, et al. Expression of mRNA and protein of macrophage colony-stimulating factor and its receptor in human follicular luteinized granulosa cells［J］. Fertil Steril, 2005, 83(2): 419-425.
［7］余蓉，陈汉平，严小芳. 单细胞实时荧光定量PCR结合比较阈值法在诊断唐氏综合征［J］. 中华医学遗传学杂志，2007, 24(2): 200-202.
［8］Tarlatzis BC, Zepiridis L, Grimbizis G, et al. Clinical management of low ovarian response to stimulation for IVF: a systematic review［J］. Hum Reprod Update, 2003, 9(1): 61-76.
［9］Xu S, Zhang Z, Xia LX, et al. Role of macrophage colony-stimulating factor (M-CSF) in human granulosa cells［J］. Gynecol Endocrinol, 2016, 32(12): 1005-1008.
［10］朱桂金，聂睿，徐蓓. 卵泡刺激素和黄体生成素阈值与阈值窗及卵泡的发育［J］. 生殖医学杂志，2008, 17(6)：401-404.
［11］Churchill SJ, Wang ET, Pisarska MD. Metabolic consequences of polycystic ovary syndrome［J］. Minerva Ginecol, 2015, 67(6): 545-555.
［12］张玲，贾莉婷，张展，等. 巨噬细胞集落刺激因子在多囊卵巢综合征大鼠卵巢中的表达及意义［J］. 中国实用医刊，2009，36（2）：9-11.
［13］Zhang Z, Fang Q, Wang J. Involvement of macrophage colony-stimulating factor (M-CSF) in the function of follicular granulosa cells［J］. Fertil Steril, 2008, 90(3): 749-754.
［14］钟明. 难治性PCOS不孕症治疗研究进展［J］. 中华全科医学，2010, 8(10)：79-81.

[1]	杜尘, 田禾, 曹阳, 王拥军, 张岩. 从内分泌激素探讨多囊卵巢综合征诱发骨代谢异常的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(3): 328-333.
[2]	欧珺, 侯文文, 唐静文, 栗家平, 徐庆阳. 靶向作用TIMP-2调控人卵巢颗粒细胞的增殖能力[J]. 中国临床药理学与治疗学, 2020, 25(5): 527-532.
[3]	郭良堂，林超，孙云，戴盛锋，张炳才，陈舒. 二甲双胍对多囊卵巢综合征患者疗效及其对患者脂肪组织miR-25水平的影响[J]. 中国临床药理学与治疗学, 2019, 24(5): 554-560.
[4]	金武敏，林佳，王佩玉，金聪聪，余蓉，赵军招. 非降调促排卵方案在多囊卵巢综合征患者行体外授精-胚胎移植治疗中的疗效比较[J]. 中国临床药理学与治疗学, 2016, 21(10): 1178-1184.
[5]	陈雅，叶碧绿，郑九嘉，林金菊，赵军招，张李雅，杨旭. 血清及卵泡液抗苗勒管激素在多囊卵巢综合征患者体外受精促排卵治疗中的价值[J]. 中国临床药理学与治疗学, 2015, 20(12): 1405-1411.
[6]	李维宏, 李斌. 阿司匹林对多囊卵巢综合征大鼠炎症反应的影响[J]. 中国临床药理学与治疗学, 2015, 20(1): 19-23.
[7]	王佩玉, 余蓉, 林金菊, 肖仕全, 金聪聪, 朱如如. GnRH拮抗剂和GnRH激动剂在多囊卵巢综合征患者IVF-ET中的应用比较[J]. 中国临床药理学与治疗学, 2014, 19(7): 795-800.
[8]	颜元良, 龚志成. 卵泡刺激素受体基因多态性对女性卵巢功能影响的研究进展[J]. 中国临床药理学与治疗学, 2013, 18(8): 944-949.
[9]	胡卫华, 徐庆阳, 唐静文. 来曲唑与枸橼氯米芬对多囊卵巢综征患者诱导排卵的效果分析[J]. 中国临床药理学与治疗学, 2013, 18(7): 789-793.
[10]	叶慧君, 江延姣, 李爱萍, 俞渊. 桂枝茯苓胶囊联合达英-35和枸椽酸氯米芬治疗多囊卵巢综合征合并不孕症的临床观察[J]. 中国临床药理学与治疗学, 2012, 17(6): 691-695.
[11]	林佳, 刘广南, 潘竞锵, 邝少松, 黄小琼, 饶子亮. 溴隐停、罗格列酮和二甲双胍对改善多囊卵巢综合征(PCOS)大鼠胰岛素抵抗和下调促炎细胞因子及调血脂作用研究[J]. 中国临床药理学与治疗学, 2010, 15(4): 391-396.
[12]	邓阿黎, 向楠, 周忠明, 姜惠中, 张亚兵. 复方中药对多囊卵巢综合征模型大鼠糖代谢的影响[J]. 中国临床药理学与治疗学, 2009, 14(5): 504-508.
[13]	邓阿黎, 周忠明, 向楠, 姜惠中, 张亚兵, 金晶, 余念, 陈露. 复方中药对多囊卵巢综合征模型大鼠血清性激素、瘦素水平的影响[J]. 中国临床药理学与治疗学, 2008, 13(10): 1139-1143.
[14]	黎平, 阮晓红, 郭江华, 陈晓燕, 邓爱民. 来曲唑治疗排卵异常的临床疗效评价[J]. 中国临床药理学与治疗学, 2006, 11(11): 1279-1284.
[15]	孙艳辉, 侯相麟, 李兴玉. 巴西蘑菇多糖对小鼠脑组织表达粒-巨噬细胞集落刺激因子的影响[J]. 中国临床药理学与治疗学, 2004, 9(7): 799-802.