| ObjectiveTo observe the level of estradiol (E2) in serum by different doses of adenine on infertile rats with kidney-yang deficiency and the localization and expression of P450arom,CYP19 and TGF-β1 in the testis of infertile rats.To investigate the pathological mechanism of spermatogenesis caused by kidney-yang deficiency.Methods32 SD male rats were averagely randomized into 4 groups: normal control group and groups treated with low (100 mg/kg daily), middle (150 mg/kg daily) and high (200 mg/kg daily) dosage of adenine. The rat model of kidney-yang deficiency infertility was induced by intragastric injection of adenine for 30 days. Symptoms, signs and serum levels of sex hormones, sperm count with automatic sperm analyzer ,blood urea nitrogen (BUN) and serum creatinine (SCr) as well as pathological changes of the testis and kidney were observed. The expression and change levels of P450arom,CYP19,TGF-β1 were measured with immunohistochemical SABC and Western blot methods. ResultsCompared with those of normal control group, levels of testosterone (T) and estradiol (E2) were significantly decreased in groups treated with low, middle and high dosage of adenine (P<0.05), levels of SCr and BUN were significantly increased (P<0.05), and the number of sperm was significantly reduced (P<0.05), while levels of follicle-stimulating hormone (FSH) and luteotrophic hormone (LH) did not show significant changes (P>0.05), but there are no significant changes in three model groups. Some pathological changes were found in the testis and the kidney of rats treated with different dosage of adenine. The array of spermatogenic cells in the testis was disorder, and spermatogenic cells became sparse. There was more connective tissue between contorted seminiferous tubules. The number of interstitial cells was obviously decreased. Germinal epithelium within the lumen became thinner. The number of sperms was decreased. Caducous spermatoblasts were found in bulk of lumens. Crystal object deposits were found in renal tubule. Foreign-body granuloma formed and occupied integral lumens. The endepidermis of renal tubule became swollen, necrotic and caducous, and renal tubule was capsularly broadened. Fibroplasia in the mesenchymal of the cortex and medulla as well as bulk of inflammatory cell infiltrate and fibrinoid necrosis were found.With the increase of adenine dose, the damage of kidney and testis was progressively increased. CYP19 was expressed in Leydig cells, elongating spermatids in rats'testis. P450arom and TGF-β1 were positive in Leydig's cells. Compared with the normal control group, the expression of TGF-β1 was significantly higher in testis of model groups (P<0.05); the expression of CYP19 and P450arom was significantly lower (P<0.05). There was no significant difference (P>0.05) in the expression of TGF-β1, P450arom and CYP19 among three model groups. ConclusionThe level of E2 in serum and sperm counts were decreased in different doses of adenine on infertile rats with kidney-yang deficiency, which accompanied the abnormal changes of P450arom, CYP19 and TGF-β1 in the model groups. It explained that estrogen and related factors may be related to the spermatogenesis with kidney-yang deficiency.
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