Expression Of Basic Fibroblast Growth Factor And Cystic Fibrosis Transmembrane Conductance Regulator On Granulosa Cells In Polycystic Ovarian Syndrome And The Research On Relationships Of The Two Factors And IVF-ET Results

ObjectivePolycystic ovarian syndrome (PCOS) is the most common endocrine disorder diseases in the women of the childbearing ages and its main feature is the androgen excess and the chronic anovulation.The incidence rate is about 5% to 10% of all women, accounting for 75% of women with no ovulating, and up to 85% of the hairy women. In 1935, Stein and Leventhal initially reported, and it is also known as Stein and Leventhal syndrome. The clinical characters of this disease are menstrual disorders, obesity, masculine performance, infertility and the bilateral ovarian polycystic changes. Because of the highly heterogeneous clinical syndrome, the difference of the pathophysiology is very significant, including of genetics, glycometabolism, neuroendocrinology, protein-metabolism, adipic-metabolism and the abnormality of local regulating and controlling factors of ovary. So far, the etiology is not very clear, because of the complex pathogenesis, so its research is still one of the focuses in Gynecological endocrinology field. Recent studies indicated that the microenvironment changes of the ovarian became the focus and the functional disorder of some local cytokines in the ovary might be the important component of PCOS. The Basic fibroblast growth factor (bFGF) and Cystic fibrosis transmembrane conductance regulator (CFTR) are two kinds of important local regulatory factors in the ovary.bFGF, which includes of 146 sequences of amino acids, is a kind of polypeptide growth factor. It has a wide range of biological function. bFGF plays the role in the process of reproduction including select and development of follicle,and improve ovulatio,the development of embryon,and so on. CFTR is a small conductance CL channel, which is saccharified highly. CFTR regulates endocrine of women, sperm capacitation and embryo implantation and so on. The research of bFGF was on the focus of ovarian tissue and the study of CFTR was on OHSS. No report about the expression of bFGF and CFTR on granulosa cells of PCOS is read. In our study, we select two groups people, one is the PCOS patients, the other is control guoup. We investigate the espression of bFGF and CFTR on granulosa cells by using immunohistochemical methods and reverse transcription polymerase chain reaction (RT-PCR), so as to discuss the function of bFGF and CFTR in the pathogenesis of polycystic ovarian syndrome.Material and MethodsFrom patients undergoing in vitro fertilization treatment by short time in vitro fertilization, we select 30 PCOS patients and 40 controls.IVF stimulation protocol:GnRHa-based protocol (TriPtorelin/Gonal-F and HCG). A transvaginal ultrasound-guided oocyte retrieval was scheduled 36-37h HCG later. Embryos were transferred 2-3 days after the oocyte retrieval. Luteal phase support was given by daily intramuscular injection of P in oil (60mg). A pregnancy test was administered 14 d and 18 d after the embryo transfer. If the test was positive, a abdominal ultrasound study was proformed 5 weeks later to confirm a clinical pregnancy. Blood samples were collected:basal, at7:00 am on the day of injection HCG. The follicular fluid (FF) (r>15mm) was taken from 30 PCOS and 40 control group subjects and centrifuged at 2000 r for10 min immediately. Granulosa cells were collected during oocyte Pick-up.Serum and Granulosa cells samples were stored at -80℃until assayed. bFGF mRNA and CFTR mRNA expression levels on luteinizing granulose cells in IVF cycles were investigated by RT-PCR. bFGF and CFTR protein stainings in luteinizing granulose cells were measured by using immunohistochemical methods. The concentrations of oestradiol(E2),Progesterone(P) and testerone(T) in follicular fluid and in serum IVF cycles were measured by full-automatic electrochemistry luminescence immunity analyzer.Results1. The expression of bFGF mRNA in luteinizing granulose cells during IVF cycles was positive by RT-PCR. The expression level of bFGF mRNA in PCOS group was higher than control group, which has significant differences(P<0.05). The immunoreactivity for bFGF was localized in luteinizing granulose cells during IVF cycles.2. Multiple regression analysis showed that bFGF levels in serum and follicular fluid correlated with estradiol (r=0.643, P<0.05)3. The expression of bFGF levels did not correlat with the outcomes in IVF-Cycles (P>0.05)4. The expression of CFTR mRNA in luteinizing granulose cells during IVF cycles was positive by RT-PCR. The expression level of CFTR mRNA in PCOS group was higher than control group, which has significant differences (P< 0.05). The immunoreactivity for CFTR was localized in luteinizing granulose cells during IVF cycles.5. Multiple regression analysis showed that CFTR levels in serum and follicular fluid correlated with estradiol and P (r=-0.672, P<0.05;r=0.457, P<0.05)6. The expression of bFGF levels did not correlat with the outcomes in IVF-Cycles (P>0.05)Conclusions1. bFGF expresses from PCOS luteinizing granulose cells during IVF cycles, suggesting that bFGF may play an important role in the follicle development through regulating E2 to affect the development of follicular.2. The expression of bFGF levels correlated with estradiol, without the outcomes in IVF-Cycles.3. CFTR expresses from PCOS luteinizing granulose cells during IVF cycles, suggesting that CFTR may play an important role in the follicle development through regulating conductance CL- channel.4. The expression of CFTR levels correlated with estradiol and P, without the outcomes in IVF-Cycles.