Analyzining The Effect Of Macrophage Colony-Stimulating Factor And Interleukin-1β In Vitro Fertilization-embryo Transfer

BackgroundWith the increase in infertility couples, human assisted reproductive technology (Assisted Reproductive Technology ART) also received a wide range of applications, In vitro fertilization-embryo transfer technology (in Vitro Fertilization-embryo Transfer IVF) is one of the representatives of technical. With the In-depth studying of Reproductive Medicine, people found that it is utilized not only by the reproductive process in the neuroendocrine system, but also by growth factors and cytokines. Macrophage colony-stimulating factor (M-CSF) is an important member regulating in the hematopoietic system and in the a cytokine network. It is widely distributed in the body and it is important not only for the development of the body on a variety of diseases, but also involved in embryonic, trophoblast cells, the regulation of follicular and other reproductive system, it plays an important impact on the reproductive process. Interleukin-1β(IL-1β), also known as lymphocyte activating factor by the people, is first found in the hypothalamus-pituitary regulation and adjust to the reproductive through a variety of ways. So far, the studies of M-CSF and IL-1βon sex hormone and progesterone in vitro fertilization focus on the follicular fluid and serum of the day oocyte retrieval, and the result is controversial, The aim of this study is to evaluate the clinical value of M-CSF and IL-1βthrough a systematic study of its dynamic changes in Vitro Fertilization-embryo Transfer and relationship on the sex hormone and the expression on granulosa cells and the outcome of progesterone.Subject and Method1. Object Choice60 patients treated by in vitro fertilization-embryo transfer (IVF-ET) were chosen from Aug.2010to Dec 2010 in our Center of Reproductive and 20 in low-response group divided by the numbers of oocytes retrieval(oocytes retrieval number≤5),20 in middle-response group divided by the numbers of oocytes retrieval(5<oocytes retrievalnumber≤15),20 in high-response group divided by the numbers of oocytes retrieval (ocytes retrieval number>15)2. Methods2.1 The relationship of M-CSF and IL-1βlevels in serum and follicular fluid and pregnancy outcomeVenous blood samples were obtained on the following days:the 3rd or 4th of a spontaneous cycle preceding IVF, the 14 days in which injection of Triptorelin was confirmed, the day of human chorionic gonadotropin (HCG) administration, the day on oocyte pick-up (OPU) and the 2nd day after oocyte pick-up(OPU48),The first tube of clear follicular fluid was also collected. Serum and follicular fluid M-CSF and IL-1βlevels were determined using enzyme-linked immunosorbent assay; FSH, LH, E2 and P were measured by measured by full-automatic electrochemistry luminescence immunity analyzer.2.2 The expression of M-CSF and IL-1βin human egg cells of granular60 patients were chosen from the 60 patients above, all of the granulosa cells were collected in follicular fluid on the day of OPU, and the expression of M-CSF and IL-1βmRNA was detected by RT-PCR and immunohistochemistry method. Results1. The levels of M-CSF, IL-1βin serum on the day of HCG administration and oocyte retrieval in pregnancy group(117.69±33.68 ng/l,107.84±36.86 ng/l; 279.46±148.60 pg/ml,170.28±99.98 pg/ml)are higher than in non-pregnant group(79.40±38.37 ng/l,74.40±38.32 ng/l; 178.30±82.29 pg/ml,126.30±110.91 pg/ml). (P<0.05); The levels of IL-1βin serum in pregnancy group(63.34±33.04 pg/ml)is lower than in non-pregnant group(129.08±119.53 pg/ml) on the day of 48 hours after oocyte retrieval (P<0.05).2. The M-CSF level in follicular fluid (956.33±257.83 ng/l; 704.84±339.07 ng/l)was significantly higher than that in serum(107.84±36.86 ng/l; 74.40±38.32) on the day of oocyte retrieval (P<0.05).3. The IL-1βand M-CSF both in serum and follicular fluid increased with the increase of amount of harvested oocyte and the E2(r>0.67) (the day of HCG administration and oocyte retrieval).4. The IL-1βand M-CSF are positively related to the E2 level in the whole pregnancy circle.5. The M-CSF and IL-1βlevels in serum of patients<30years(121.41±23.37 ng/l;277.15±65.77pg/ml) were significantly higher than that>30years(82.35±20.34 ng/l;177.99±61.62 pg/ml) on the day of HCG administration(P<0.05); The M-CSF and IL-1βlevels in serum of patients application of gonadotropin dose≤2250IU(112.84±35.54 ng/l;263.40±132.65 pg/ml) were significantly higher than it> 2250IU(76.02±33.12 ng/l;178.72±84.47 pg/ml) (P<0.01)6. On the day of HCG adminitration, the M-CSF and IL-1βlevel in serum from low response group, the reaction group and high response group was gradually increasing trend (49.95±15.62 ng/ml,98.57±23.85 ng/ml,137.14±18.47 ng/ml; 122.39±46.80 pg/ml,220.32±89.54 pg/ml,325.30±136.43 pg/ml).he difference was statistically significant(P<0.05).7. The expression of M-CSF and IL-1βmRNA and Protein in low response group, the middle reaction group,and high response groups is from low to high order(0.134±0.027,0.502±0.113,0.833±0.1; 0.153±0.046,0.338±0.072 0.619±0.136),the difference was statistically significant(P<0.015).Conclusion1. The M-CSF and IL-1βin serum on the day of HCG and the oocytes can promote embryo implantation, and thus increase the pregnancy rate;The higher of IL-1βafter oocyte retrieval 48 hours can inhibit embryo implantation, reduced pregnancy rate2. The M-CSF, IL-1βin serum can be an indicator for predicting pregnancy outcome.