| Objective By measuring vascular endothelial growth factor (VEGF)levels in both serum on the day of human chorionic gonadotrophin (HCG) administration and embryo transfer and follicular fluid on the day of oocyte harvest of ovarian hyperstimulation syndrome (OHSS)patients and non -OHSS patients, and by studying VEGF mRNA expression, We try to explore the role of VEGF in pathogenesis of OHSS so as to provide experimental evidence for early diagnosis of OHSS.Methods 1: 14 moderate or severe OHSS patients and 22 non-OHSS patients were enrolled in this experiment. Serum was collected on the day of HCG injection and embryo transfer and follicular fluid was collected on the day of oocyte harvest from both group of OHSS and non-OHSS in vitro fertilization -embryo transfer (IVF-ET) patients. VEGF levels were measured by enzyme-labeled Immunsorbent assay. The results were shown in X 盨, t test were used in statistical analysis. 2:The expression of VEGF mRNA of granulosa cells was detected by RT-PCR. Granulosa cells were collected on the day of oocyte harvest. Total RNA of them was prepared and calculated according to formula A260 40 diluent times/1 OOO.cDNA was synthesized by reverse transcription (RT).The total volume of reaction system was 20ulincluded 4uL MgCL2, 2ul IQxRNAPCRbuffer, 2ul dNTP mixture, O.Sul Rnase Inhibitor, lul Random 9mers, lul Reverse Transcriptase, lug sample. The reaction condition was :30 10min, 42 30min, 99 5min, 5 5min Following RT reaction , 6uL MgCL2, 8ul 10XRNA PCR buffer, 61.5ul pure water , 0.5ul TaKaRa Taq, lul of VEGF upstream and downstream primer respectively, lul of P -actin upstream and downstream primer respectively were put in. The reaction condition was: 94 Imin, 62 1min, 72 Imin, 28 cycles/72 7 min. PCR product was separated by 1.5%sepharose electrophoresis. The results were analysed by Alpha-Imager.Results 1 The results of ELISA show as following: There is no significance difference between two groups on levels of VEGF on HCG injection day in serum (102.5+40.80 and 90.23+34.62 pg/ml respectively,P>0.05); There is significance difference between two groups on levels of VEGF on ET day in semm( 150.93+59.91 and 104.18+20.76 pg/ml respectively, P<0.01); There is significance difference between two groups on levels of VEGF in follicular fluid, they were782.86+136.87 and 362.50+231.85 respectively, P<0.05. 2 The comparison of Serum VEGF on HCG injection day and embryo transfer day shows significance difference in OHSS group (102.5+40.80 and 150.93+59.91 pg/ml respectively, P<0.05);but no significance difference in non-OHSS group (90.23+34.62 and 104.18 +20.76 pg/ml respectively, P>0.05). 3 VEGF levels in follicular fluid were several times higher than that in serum (782. 86+136. 87 versus 150.93+59.91,102.5+40. 80 pg/ml, 362. 50+231.85versus 104.18+20. 76 90. 23+34. 62pg/ml) .4 The results of RT-PCR revealed that the ratio of VEGF/ P -actin in OHSS group and non OHSS group were 0.54+0.24 and 0.27+0.12 respectively, shows significant difference between two groups (PO.01). The expression of VEGF mRNA in OHSS group was stronger than that in non-OHSS group.Conclusion 1 VEGF level increase in serum on embryo transfer day and in follicular fluid on oocyte-harvest day was more obvious in OHSS group than that in non-OHSS group. The expression of VEGF mRNA also shows stronger in OHSS group. The levels of VEGF in serum on embryo transfer day and in follicular fluid on oocyte-harvest day can be used to predict OHSS occurance. 2 VEGF levels in serum on embryo transfer day was higher than that in HCG injection day in OHSS group, but not in non-OHSS group. The rise of VEGF levels after HCG administration might be used as an item in early diagnosis of OHSS occurance. 3 VEGF levels in follicular fluid were several times higher than that in serum .This means that in the pathophysiological process of OHSS, the expression of VEGF in the ovary become more intense. 4 There were no significant difference of VEGF levels on HCG injection day between OHSS group and non-OHSS group. This indicat...
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