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Objective: To establish a rat model of experimental varicocele (varicocele, VC), by measuring the testicular tissue androgen-binding protein, inhibin B expression changes, and the concentration of testosterone in the testicular tissue to understand the VC of the Sertoli cells secrete the damaging effects of the function, and secretion of endogenous testosterone testicular tissue to explore the mechanism of VC lead to infertility. Method: Select the mice age (50 ± 10) days (puberty), weighing 240 ~ 280g SD (Sprayue-Dawley) male rats 40. The rats were divided into the varicocele group (experimental group) 20, 20 of the sham group (control group) principle of randomization. The animal model is established based on Turner's method: the experimental group rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (3.33 ml / kg), the abdominal midline incision approximately 5cm incision full thickness abdominal wall, exposed to the abdominal cavity. Left renal vein, left adrenal central vein, left spermatic vein location is confirmed, the careful separation of the left renal vein between the center of the left adrenal vein and the inferior vena cava, left renal vein placed a diameter of about 0.8 mm metal rod parallel with 4-0 silk ligation of the left renal vein with venous diameter is approximately reduced by half. Ligation out after the completion of the metal rod, visible to the rapid expansion of the left renal vein. With 4-0 silk suture wounds. Control rats only wear line knotted to expose the left renal vein, without narrowing of the left renal vein. After 12 weeks, the rats were killed by cervical dislocation after taking abdominal incision, based on the maximum diameter of the spermatic vein gt; 1 mm left and right kidney weight was no significant difference in success criteria for modeling pushed into the abdominal cavity, and then on the left testicle observe changes in testicular size, testicular epididymis, cut, carefully cut off the epididymis and surrounding fascia and other affiliated organizations, washed with cold saline, filter paper moisture. Testicular average cut in half, half cut off albuginea weighed and put into a glass homogenizer, adding phosphate buffer after grinding ratio of 1:2 (weight: volume), 4 ℃ overnight, the next day to 3000 rev / min, centrifugal 15min, the supernatant was placed Eppendof tube at -20 ℃ under test. The other half of the testicular tissue was fixed in 10% neutral buffered formalin solution 24 hours, after gradient dehydration, paraffin-embedded tissue blocks. The testicular tissue androgen binding protein, inhibin B was measured by tissue sections and immunohistochemical staining methods. Testis testosterone concentrations were measured using Access Testosterone (chemiluminescence method), BECKMAN COULTER DX Ⅰ 800 automated immunoassay analyzer and determination of sample concentration of testosterone in strict accordance with the provisions of processes. Measurement data were x_ ± s said that the number of samples using two independent samples t-test, nonparametric tests using the Wilcoxon rank sum test, using SPSS 13 0 software for statistical analysis, the use of two-sided test, P lt; 0.05 statistically significant. Results: 1 testicular tissue androgen binding protein measurement results: after 12 weeks, measured by immunohistochemistry method, the experimental group rat the left testis tissue androgen binding protein (ABP) immunohistochemical score (IHS) the Median = 6.00 , P25 = 4.00, P50 = 6.00, P75 = 8.25, control group the IHS rated the Median = 6.00, P25 = 4.50 The P50 = 6.00, P75 = 9.00, Wilcoxon rank sum test was no significant difference, P gt; 0.05 2 testicles inhibin B measurement results within the organization: after 12 weeks, the same determined using immunohistochemical methods, experimental rats the left testicular inhibin B (InhB) immunohistochemical score (IHS) is the Median = 5.00, P25 = 4.00, P50 = 5.00, P75 = 6.00, control group the IHS rated the Median = 7.50, P25 = 4.50 The P50 = 7.50, P75 = 9.00, and the control group experimental group compared InhB expression reduced rank sum test differences statistically significant P lt; 0.05 3 testicular tissue testosterone levels Determination: After 12 weeks, the testosterone concentration measured by Access Testosterone testicular tissue homogenates extract, the experimental rats left testis tissue homogenates extract the concentration of testosterone for the control group, 94.79 ± 3.31nmol / L, 109.10 ± 6.37nmol / L, testosterone levels were significantly decreased, compared with the control group, a statistically significant difference, P lt; 0.05 4 pathological changes of testicular damage: testicular tissue damage The outstanding performance of the pathological changes in seminiferous tubules varying degrees of spermatogenic cell shedding, pathological changes of the experimental group SD rat testis, spermatogenic cell shedding, severe cases can occur lumen blockage, but did not find the CD. support cell syndrome and other serious pathological changes. 2, visible limiting membrane thickening, fibrosis and hyalinization;, testicular interstitial edema, interstitial cell degeneration (cytoplasmic vacuolar degeneration); 4, the Leydig vascular pathological change, including small artery lesions (wall thickening, luminal narrowing or occlusion) small venous dilatation and congestion and lymphangiectasia. Conclusion: after varicocele testicular androgen-binding protein expression is not significantly reduced. 2 varicocele Sertoli cells inhibit cytochalasin B decreased production, and its weakened role of mesenchymal cells. 3 varicocele reduce the level of testosterone in the testicular tissue, affect sperm development, maturation, leading to male infertility.
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