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目的:通过测定无精子症患者睾丸组织中内皮型一氧化氮合酶运输介导物(NOSTRIN)和内皮型一氧化氮合酶(eNOS)的表达,探讨其与无精子症发病的相关性。方法:采用免疫组化法检测17例特发性无精子症患者(病例组)和10例正常男性(正常组)睾丸组织中NOSTRIN的定位和定量表达;RT-PCR检测睾丸组织中NOSTRINmRNA的表达;分光光度法测定睾丸组织中eNOS的活性;应用硝酸还原酶法测定睾丸组织上清液中NO代谢产物亚硝酸基/亚硝基(NO2-/NO3-)水平。结果:NOSTRIN在睾丸组织中表达于生精细胞、支持细胞以及血管内皮细胞,NOSTRIN在特发性无精子症患者睾丸中的表达水平显著低于正常男性;病例组患者睾丸组织中NOSTRINmRNA呈弱表达(0.312±0.076),明显低于正常组(0.793±0.082,P<0.01);病例组患者睾丸组织eNOS活性[(33.727±3.58)U/mg]与正常组[(17.69±3.84)U/mg]比较显著增高(P<0.01);病例组患者睾丸中NO2-/NO3-水平为[(48.56±8.49)μmol/L],与正常组[(25.37±9.61)μmol/L]比较显著升高(P<0.01);病例组睾丸组织中NOSTRIN表达水平与eNOS活性呈负相关(r=-0.57,P<0.01),与NO代谢产物NO2-/NO3-同样呈负相关(r=-0.61,P<0.01)。结论:无精子症患者睾丸组织中NOSTRIN表达水平降低,eNOS活性增强,NO-/NO-水平升高,这可能与无精子症的发生有着相关性。
OBJECTIVE: To investigate the relationship between the expression of endothelial nitric oxide synthase transporter (NOSTRIN) and endothelial nitric oxide synthase (eNOS) in testicular tissue of azoospermic patients and explore its relationship with the development of azoospermia. Methods: The localization and quantitative expression of NOSTRIN in 17 cases of idiopathic azoospermia (case group) and 10 cases of normal male (normal group) were detected by immunohistochemistry. The expression of NOSTRIN mRNA in testis tissue was detected by RT-PCR ; Determination of eNOS activity in testis tissue by spectrophotometry; NO metabolite nitrite / nitrite (NO2- / NO3-) levels in testis tissue supernatant was determined by nitrate reductase method. Results: NOSTRIN was expressed in spermatogenic cells, supporting cells and vascular endothelial cells in testis tissue. The expression of NOSTRIN in testes of patients with idiopathic azoospermia was significantly lower than that in normal males. The expression of NOSTRINmRNA in testis was weak (0.312 ± 0.076), which was significantly lower than that of the normal group (0.793 ± 0.082, P <0.01). The eNOS activity in the testicular tissue was significantly higher than that in the normal group [(33.727 ± 3.58) U / mg and (48.56 ± 8.49) μmol / L], which was significantly higher than that in the normal group [(25.37 ± 9.61) μmol / L] (P <0.01). There was a negative correlation between the expression of NOSTRIN and the activity of eNOS in testis (r = -0.57, P <0.01), and also negatively correlated with NO metabolites NO2- / P <0.01). CONCLUSIONS: The expression of NOSTRIN, the activity of eNOS and the level of NO / NOS in testicular tissue of azoospermia patients may be correlated with the development of azoospermia.