Impaction Of Hydrosapinx On Endometrial Receptivity

Objective: To study the impaction of hydrosapinx on the outcome of IVF-ET. To explore gene expression changes of hydrosapinx patient in the window of implantation (WOI), demonstrate differentially expressed genes, and explore their clinical significance. Validation of array data was accomplished by mRNA quantification by real time quantitative fluorescent PCR (RT-PCR) and by level of protein expression of some differentially expressed genes. And explore their clinical meaning.Methods: Statistic Analysis: Two groups composed of patients between August 2003 and August 2005 are built. Hydrosapix group is patients with hydrosapinx treated with IVF-ET for 143 cycles. Tubal obstruction group is patients with other pathologic change without hydrosapinx treated with IVF-ET for 263 cycles. Compare the two groups' IVF-ET outcome.Oligonucleotide Microarray : The differentially expressed genes (Signal Log Ratio (?)2 or (?)2) of endometrium were analyzed among hydrosapinx, tubal obstruction without hydrospinx and healthy volunteer in WOI, using Affymetrix U133A 2.0 gene chip, Functional classification of differentially expressed genes were established by searching in the human data-base.RT-PCR : Osteopontin (OPN or SPPI), Placenta protein-14 (PP-14, Glycodelin, GdA or PAEP) and Integrinβ₃, which is differentially expressed or closely related with endometrium receptivity, were verified by PT-PCR. The mRNA quantification of 31 hydrosapinx patients, 29 tubal obstruction patients without hydrosapinx and 21 healthy volunteers were analyzed.Immunohistochemistry: Analyze and verify the changes of three groups' OPN, GdA and Integrinβ₃ by Immunohistochemistry.SPSS 13.0 statistic package was used and p<0.05 was adopted as significance level.Results:1. The treatment outcome of IVE-ETThe quality of transferred embryo between hydrosapinx and tubal obstruction group has no statistical difference. In the hydrosapinx group, the rates of clinical pregnancy, embryo implantation, early abortion and ectopic pregnancy were 32.87%, 18.61%, 19.15% and 17.02%. In the bubal obstruction group, the rates of clinical pregnancy, embryo implantation, early abortion and ectopic pregnancy were 49.8%, 31.87%, 6.87% and 4.58%.2. Oligonucleotide Microarray:(1) The differentially expressed genesThere were 588 genes significantly differentially expressed between hydrosapinx patient and healthy volunteer, 180 genes between tubal obstruction patient and healthy volunteer, 750 genes between hydrosapinx and tubal obstruction patient without hydrosapinx. Those genes were classified into 10 groups according to their main function. They are enzyme, cell adhesion, structural protein, regulation of cell cycle, ion binding protein, signal-transporting proteins, immunomodulators, carrier proteins, unknown function ect.(2) Differentially expressed genes between hydrosapinx patient and healthy volunteerTotal of 588 genes were significantly differentially expressed. 351 genes were up-regulated, and 237 genes were down-regulated. Enzyme related genes were at most among both up-regulated and down-regulated genes. Signal-transporting proteins were second.(3) Differentially expressed genes between tubal obstruction patient without hydrosapinx and healthy volunteerTotal of 180 genes were significantly differentially expressed. 106 genes were up-regulated, and 74 genes were down-regulated. Enzyme related genes were at most among significantly differentially expressed genes. Signal-transporting proteins were next.(4) Differentially expressed genes between hydrosapinx and tubal obstruction patientTotal of 750 genes were significantly differentially expressed. 302 genes were up-regulated, and 448 genes were down-regulated. Signal-transporting proteins were the most among significantly differentially expressed genes except Enzyme.3.RT-PCR(1) The different expression of OPN genes mRNA among three groupsThe OPN relative expression quantification among hydrosapinx group, tubal obstruction group and healthy control group were 0.6238±0.2690, 0.8955±0.3887 and 1.1065±0.7821. There was no obvious difference between any two groups.(2) The different expression of Integrinβ₃ genes mRNA among three groupsThe Integrinβ₃ relative expression quantification among hydrosapinx group, tubal obstruction group and healthy control group were 0.1945±0.1024, 0.8285±0.1945 and 1.2541±0.5245.There was obvious difference between hydrosapinx group and tubal obstruction group. So was hydrosapinx group and health control group.(3) The different expression of GdA genes mRNA among three groupsThe GdA relative expression quantification among hydrosapinx group, tubal obstruction group and healthy control group were 2.0815±0.3425, 3.1578±0.6712 and 4.1257±0.9476. There was obvious difference between hydrosapinx group and health control group. So was hydrosapinx group and health control group.4. Immunohistochemistry(1) The different expression of OPN among three groupsOPN was situated in endometrium glandular epithelium and cavitas epithelial cells. There was little expression in stroma-cell. There was no significant difference among three groups.(2) The different expression of Integrinβ₃ among three groupsIntegrinβ₃ was situated in endometrium glandular epithelium and (or) cavitas epithelial cells. There was only a little expression in stroma-cell. The expression of Integrinβ₃ in most hydrosapinx patients' endometrium is absent or weakly positive. The intension of expression in hydrosapinx group is obviously lower than tubal obstruction group and healthy control group.(3) The different expression of GdA among three groupsGdA was situated in endometrium glandular epithelium and (or) cavitas epithelial cells. There was little expression in stroma-cell. Both hydrosapinx group and tubal obstruction group are obviously lower than healthy control group.Conclusion:1. Hydrosapinx patient treated with IVF-ET has low clinical pregnancy rate, while high early abortion rate.2. Hydrosapinx has influence on many endometrium genes' expression. The most differentially expressed genes are enzyme related genes and signal-transporting protein genes.3. Hydrosapinx has no effects on OPN gene expression, transcription, protein synthesis and secretion in WOI endometrium. So does tubal obstruction4. Hydrosapinx inhibits Integrinβ₃ gene expression, transcription, protein synthesis and secretion in WOI endometrium. So that destroys the ability of endometrium recepting embryo adhesion and implantation.6. Hydrosapinx inhibits GdA gene expression, transcription, proteinsynthesis and secretion in WOI endometrium. And destroys the immune tolerance of endometrium to embryo.