Influence Of Genital Tract Obstruction And Reconstruction On Epididymal Protein

Objectives:To estimate the application of microsurgery in obstructive azoospermia and to determine the relationship among obstructive interval, patency rate, semen analysis and pregnancy rate.Patients and Methods:We undertook a retrospective analysis of 35 patients treated by microsurgical procedures performed between January 2005 and May 2009. Obstructive interval was compared with patency rate, semen analysis and pregnancy rate.Results:The overall patency rate and the overall pregnancy rate were 94.3% and 51.4% respectively. The overall patency rate was significantly higher than the overall pregnancy rate. The patients whether to make his wife pregnant after surgery were divided into pregnancy and non-pregnancy group. Semen volume(t=2.0823, P<0.05), sperm count (t=2.7147, P<0.05), sperm mobility (t=2.1982, P<0.05) and normal morphology sperm rate(t=6.6762, P<0.01) of the pregnancy group were significantly higher than the non-pregnancy group, respectively. Age, spouse age and obstructive interval in pregnany group and non-pregnancy group were no significant difference. According to obstructive interval less or more than 10 years, the patients were divided into two groups. Semen volume (t=5.7390, P<0.01), sperm count(t=5.9538, P<0.01), sperm mobility (t=3.6776, P<0.01) and normal morphology sperm rate (t=3.9357, P<0.01) of the group with obstructive interval less than 10 years were significantly higher, respectively. Logisti regression analysis showed that sperm count (OR:1.1079, 95% CI:1.0157-1.2084, P<0.05), sperm viability (OR:1.1094,95% CI: 1.0000-1.2308, P<0.05) and normal sperm morphology rate (OR:1.4805,95% CI: 1.0574-2.0729, P<0.05) were significant on the recovery of fertility.Conclusion:(1) This study confirms the effectiveness of microsurgery for obstructive azoospermia men who wish to father children. (2)After microsurgery, the natural pregnancy rate was significantly lower than the rate of re-canalization. (3) Semen volum, perm count, sperm motility, normal morphology sperm and pregnancy rate decreases as the obstructive interval extends. Sperm motility and normal morphology sperm rate play a key role on fertility recovery after vas reversal. Objective:To explore the method of establishing a stable model of obstructive azoospermia and reconstruction of the genital tract in Wistar rats and to study its influence on spermatogenic function of testes.Methods:Nighty male rats were randomly divided into six groups, each group had 15 rats with the average weight of (250±20) g. With intraabdominal anesthesia by chloral hydrate, we made obstructive azoospermia models by intraabdominal vasoligation. The testes of groupⅠtoⅢwere harvested 1 month,2 months and 3 months after suegery respectively. After hematoxylin and eosin stain, the pathology of testes under light microscope was performed. With intraabdominal anesthesia by chloral hydrate, the groupⅣtoⅥunderwent transabdominal six-needle full-thickness vasovasostomy 1 month,2 months and 3 months after vasoligation, respectively. After hematoxylin and eosin stain, the pathology of testes under light microscope was performed. Microsurgery was performed with Laica microscope of 10-time amplificatioin.Results:None of rats died after operation. The pathology of testes in all groups showed multiple layers of spermatogenic cells including sperm covering on seminiferous tubule. We observed the patency rates of three genital reconstruction groups by vasography. Bilateral patency and unilateral patency of the groupⅣtoⅥwere 12 and 3,10 and 2,8 and 2, respective. The total patency rate of three group was 27/30 (90%), (73.3%) and 18/30(60%) respectively. The statistical differences of patency rates were analyzed by chi-square test, the patency rate of groupⅣis significantly higher than groupⅥ(P= 0.015). The patency rate was significantly difference among three groups (x 2=7.13, P=0.028)..Conclusion:(1) Spermatogenesis was not impaired obviously on histopathological examination by obstruction of genital tract. (2)The patency rate of obstructive azoospermia after genital tract reconstruction decreases along with the obstructive interval. (3)We establshed a stable, homogeneous and repetitive obstructive azoospermia and genital tract reconstruction model in Wistar rats by transabdominal vasoligation and six-needle, full-thickness suture. This model can help to research the pathophysiology of obstructive azoospermia and genital tract reconstruction of obstructive azoospermia.Objective:To study the influence of genital tract obstruction on epididymal protein and explore the possible mechanism of the influence on sperm maturity and fertility caused by genital tract obstruction.Method:Rat models of genital tract obstruction were established and the epididymus were harvested after obstructive interval of 1 month,2 months and 4 months respectively. Two-dimensional gel electrophoresis was used to obtain the differentially expressed epididymal proteins. Candidate proteins were determined by mass-spectrometric technique and multi-database search. Differentially expressed proteins were identified by Western Blotting and the expression regulation was analysed by Realtime PCR.Results:(1) Two-dimensional gel electrophoresis of protein showed change of HE2, fertilin, FA-1, Clusterin, CRISP1 and ERABP after genital tract obstruction and there was difference among samples of different obstructive interval. (2) HE2, FA-1 and Clusterin were verified by Western Blotting. These proteins significantly decreased after obstruction. (3) The result of Realtime PCR revealed that the mRNA transcription of HE2 gene was influenced by genital tract obstruction.Conclusion:(1) Genital tract obstruction leaded to change of epididymal function which was showed by the decrease of epididymal functional proteins. (2)The content of HE2, Fertilin, FA-1, Clusterin, CRISP1 and ERABP which could promote sperm muturity and fertility deceasing along with the obstructive interval indicates that the fertility of sperm due to genital tract obstruction is caused by decrease of epididymal proteins. (3)The content change of HE2 after genital obstruction is caused by gene expression indicating that gene expression could be regulated by genital tract obstruction. Objective:To study the influence of re-canalization of genital tract obstruction on epididymal proteins. To explore the possible mechanism of the influence on sperm maturity and fertility caused by re-canalization of genital tract obstruction and the time-related effect for genital tract reconstruction.Method:Rat models of genital tract obstruction were established and genital tract construction was perfermed after 1 month,2 months and 4 months, respectively. The epididymus were harvested 1 month after genital tract reconstruction. Two-dimensional gel electrophoresis was used to harvested the differentially expressed epididymal proteins. Candidate proteins were determined by mass-spectrometric technique and multi-database search. Differentially expressed proteins were identified by Western Blotting and the expression regulation was analysed by Realtime PCR.Results:(1) Two-dimensional gel electrophoresis of protein showed change of protein compositons and content after genital tract obstruction. Hyaluronidase PH-20 protein remained unchanged. Fertilin, FA-1, CRISP1 and ERABP continued to decline. HE2 and Clusterin levels partially recovered. And there were differences among genital tract reconstruction samples of different obstructive interval. (2) Hyaluronidase PH-20 did not significantly change after genital tract reconstruction varified by Western blot and RT-PCR. (3)The result of Western blot and Realtime PCR revealed that the HE2 gene was regulated on the level of mRNA transcription by genital tract reconstruction and the HE2 content recovered in a certain extent. The recovery was obvious after short obstructive interval (1 month and 2 months), but recovery did not happened after long obstructive interval (4 months).Conclusion:(1) It is possible that genital tract reconstruction could lead to recovery of epididymal function to a certain extent. The shorter the interval of genital tract obstruction, the better the epididymal function recovers. (2) Time-effect relationship between epididymal protein restoration and obstructive interval could explain time-effect relationship between fertility restoration after genital tract obstruction and obstructive interval. HE2 proteins may play an important role in fertility restoration.