| ObjectiveBefore male sexual maturity,high ligation of unilateral spermatic vessels has been extensively used in the treatment of some urinary system diseases,such as Fowler-Stephens operation for high cryptorchidism and Palomo operation for varicocele.However,since this type of operation cuts off the spermatic vessels,it may cause acute ischemia on the affected side of the testis.Therefore,it is extremely important to evaluate the function of the testis regularly after the operation.Due to the poor compliance of children with long-term follow-up after operation and the high rate of loss to follow-up,it is difficult in obtaining sufficient data for long-term evaluation of testicular function after sexual maturity,so there are few studies in clinical on the long-term effects of testicular function after sexual maturity about high ligation of unilateral spermatic vessels before sexual maturity.Therefore,since the characteristics of SD rats’ testicular development are similar to those of humans,this study established a model of unilateral spermatic vessels ligation in SD rats before sexual maturity,and regarded analysis of the pathology,serum sex hormone levels and semen quality of the testis after sexual maturity as the main long-term evaluation index of testicular function.This paper preliminarily explored the effect of unilateral spermatic vessels high ligation before sexual maturity on the long-term testicular function after sexual maturity in SD rats.It provides basic research basis and data support for the clinical evaluation of the long-term impact of unilateral spermatic vessels ligation on testicular function.MethodsSixteen 30-day-old healthy male SD rats were randomly divided into the experimental group of unilateral spermatic vessels ligation and the sham operation control group,with 8 rats in each group.An animal model of unilateral（right）spermatic vessels ligation was established by surgery.After successful modeling,the rats continued to be raised for 30 days.At 60 days of age,venous blood were drawn under anesthesia,and serum LH,FSH and T levels were detected by ELISA method;bilateral testicular tissues were taken,fixed,embedded,sectioned and HE stained,pathological changes of testicular tissue were observed under microscope,and testicular weights were recorded;bilateral epididymis tissues were taken and made into sperm suspension,then sperm density,viability and abnormality rate were measured under microscope,and the data obtained were analyzed statistically.ResultsWe made successful establishment of unilateral spermatic vessels ligation models and sham operation models respectively in two groups.After modeling,the rats continued to be raised to 60 days of age,and reached sexual maturity.Observation of the general morphology of the testes found that,the testes on the operation side of the experimental group shrank to different degrees,and the morphological size was reduced to different degrees compared with the non-operation side of the experimental group;the testes on the operation side of the control group were not significantly different from those on the non-operation side of the control group.Testicular weight comparison results: the weight of the testes on the operation side of the experimental group（2.26±1.63g）was smaller than that of the non-operation side of the experimental group（4.09±0.40g）,and the difference was statistically significant（P=0.02）;but no significant difference was found between the weight of the testes on the operation side of the control group（3.97±0.26g）and the non-operation side of the control group（3.98±0.31g）;the weight of the testes on the operation side of the experimental group was smaller than that of the operation side of the control group,and the difference was statistically significant（P=0.02）;but no significant difference was found between the weight of testes on the non-operation side of the experimental group and the non-operation side of the control group.HE staining observation revealed that,the experimental group had different degrees of pathological changes in the testes on the operation side,the number of abnormally structured seminiferous tubules increased,the number of seminiferous epithelial layers decreased,and only spermatogonia could be seen on part of the seminiferous tubule walls,some tubules were atrophy,collapse,or in irregular shape,a small amount of parenchymal necrosis and calcification were seen locally;one case showed severely atrophy of the testis on the operation side,the shape of the seminiferous tubules in the testis basically disappeared,large areas of parenchymal cell necrosis,nucleus fragmentation or dissolution,the interstitium was indistinguishable,the tissue structure was disordered,all kinds of inflammatory cell infiltrated,accompanied with connective tissue hyperplasia and granuloma formation,calcification foci and calcium salt deposits could be seen near the center of the testis;the morphology and structure of testes on the non-operation side of the experimental group,the operation side of the control group and the non-operation side of the control group were all normal.ELISA test results are as follows,serum LH concentration: the experimental group（172.37±14.27 mIU/ml）was higher than the control group（164.68±13.38 mIU/ml）;serum FSH concentration: the experimental group（47.56±30.25 mIU/ml）was lower than control group（51.02±20.32 mIU/ml）;serum T concentration: the experimental group（50.99±5.04nmol/L）was lower than the control group（59.47±20.71nmol/L）.There was no statistical difference in LH,FSH and T between the two groups（P>0.05）.The results of semen analysis are as follows,sperm density on the operation side: the operation side of the experimental group（12.50±15.91×106/m L）was lower than the operation side of the control group（45.00±29.06×106/m L）,the difference was statistically significant（P=0.015）;sperm motility on the operation side: the operation side of the experimental group（4.88±4.79%）was lower than the operation side of the control group（17.25±12.85%）,and the difference was statistically significant（P=0.031）;the rate of sperm deformity on the operation side: the operation side of the experimental group（21.63±11.45%）was higher than the operation side of the control group（14.88±6.27%）,but there was no statistically significant difference（P>0.05）.Sperm density on the non-operation side: the non-operation side of the experimental group（57.25±39.45×106/m L）was higher than the non-operation side of the control group（57.25±39.45×106/m L）;sperm motility on the non-operation side:the non-operation side of the experimental group（22.50 ±7.95%）was higher than the non-operation side of the control group（20.38±11.80%）;the rate of sperm deformity on the non-operation side: the non-operation side of the experimental group（14.50±5.93%）was higher than the non-operation side of the control group（9.13±4.22%）,there was no significant difference in sperm density,motility and deformity rate between the non-operation side of the experimental group and the non-operation side of the control group（P>0.05）.Conclusions1.The testes on the operation side of the experimental group showed different degrees of atrophy and pathological changes.The spermatogenic cells fell off,the number of seminiferous epithelium decreased,the lumen of the seminiferous tubules shrank and collapsed,and the local parenchymal cells of some tissues were necrotic and calcified.The morphology and structure of the testes on the non-operation side of the experimental group are normal.These showed that the high ligation of unilateral spermatic vessels in rats before sexual maturity may have varying degrees of long-term influence on the morphology and structure of the testis on the operation side,but has no effect on the morphology and structure of the testis on the non-operation side.2.There was no significant difference in serum sex hormones between the experimental group and the control group,indicating that the high ligation of unilateral spermatic vessels before sexual maturity did not affect the long-term serum sex hormone levels in rats.3.The sperm density and vitality of the epididymis on the operation side of the experimental group decreased significantly,the sperm density,vitality and deformity rate of the epididymis on the non-operation side of the experimental group were not significantly different from those of the control group,indicating that the high ligation of unilateral spermatic vessels before sexual maturity may have a long-term impact on the quality of the epididymal semen on the operation side,but has no impact on the quality of the non-operation epididymis semen.This provides a certain basic research basis and data support for the clinical long-term impact of this type of operation on testicular function. |
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