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为探讨大剂量吡哆醇 (PN)对睾丸结构和功能损害的分子机制 ,研究其对睾丸一氧化氮合酶(NOS)和细胞凋亡的影响。实验采用吖啶橙荧光染色法显示睾丸细胞DNA和RNA ,黄递酶 (NADPH -d)组织化学法显示睾丸细胞NOS ,采用原位缺口平移技术检测睾丸生精细胞凋亡的数目。结果表明 ,与对照组相比 ,注射PN 3d ,大鼠睾丸间质细胞NOS反应活性轻度增加 ;注射PN 7d后 ,大鼠睾丸细胞DNA和RNA荧光显色明显减弱 ,间质细胞NOS活性明显增强 ,睾丸生精细胞凋亡数目 (38 41± 8 2 0 )较相应对照组 (11 39± 4 86 )明显增多 ,差异有显著性 (P <0 0 1)。提示大剂量PN腹腔注射后 ,大鼠睾丸细胞NOS活性和凋亡细胞数目明显增加 ,这些变化可能是PN损害睾丸结构和功能的分子机制之一
To investigate the molecular mechanism of high dose of pyridoxine (PN) on the structure and function of testis, the effects of PN on testis nitric oxide synthase (NOS) and apoptosis were studied. Acridine orange fluorescence staining was used to detect the DNA and RNA of testis cells. NADPH-d histochemical method was used to detect testicular cell NOS. The number of spermatogenic cells apoptosis was detected by in situ nicking shift assay. The results showed that, compared with the control group, the NOS reactivity of testicular stromal cells was slightly increased after PN 3d injection. Fluorescence staining of DNA and RNA of testicular cells was significantly attenuated and the NOS activity of interstitial cells was significantly increased (38 41 ± 8 2 0) than the corresponding control group (11 39 ± 4 86), the difference was significant (P 0 01). Prompted high-dose intraperitoneal injection of PN, the rat testicular cell NOS activity and the number of apoptotic cells increased significantly, these changes may be one of the molecular mechanisms of PN damage testicular structure and function