Production of nitric oxide by Sertoli cell and its influence upon the cells

从未成熟大鼠分离的大鼠生精上皮细胞（Sertoli细胞：SC）在双室系统中或在15 mm圆形盖玻片上培养，以验证一氧化氮（NO）对SC蛋白分泌的影响和SC单层形成的渗透屏障（紧密连接）的功能。研究结果如下：1. SC在培养过程中自发产生的NO量相对较少.在培养系统的基础储库中加入1 μg/ml的LPS并不能显著增加条件培养液中NO的浓度.加入NO供体（NOC 18）释放的外源性NO对SC单层的电阻（ER）没有影响. SC分泌的总转铁蛋白（TF）的量在对照组和经NOC 18处理的上皮片层之间没有显著的统计学差异。然而，SC分泌TF的极性在NO-do时显著降低， 关于我们 也不包含文化。在2天的收集期内，在添加外源性NOC 18（50和100 μM）的培养物中，TF的分泌比（顶/底=1.86 ± 0.22）分别显著下降至1.26 ± 0.04和1.47 ± 0.08.在未处理的SC中，闭合蛋白和ZO-1的免疫染色显示在细胞-细胞连接复合物区域的细胞周边周围的连续标记，并且这些标记与TJ相关的F-肌动蛋白抑制剂共定位。然而，用NOC 18（200 μM和400 μM）孵育在48小时内导致闭合蛋白染色的完全瓦解和ZO-1染色的部分破坏。此外，occludin，ZO-1和corpendin的表达下降，观察在NO供体处理的SC的NOC 18的剂量依赖性的方式。这些结果表明，NO产生过度睾丸内急性或慢性病理生理条件下破坏TJ相关蛋白的SC和NO可能会扰乱血睾屏障的形成，反过来调节正常的精子发生。少

Rat seminiferous epithelial cells (Sertoli cells : SC) isolated from immature rats were cultured either in the bicameral chamber system or on 15-mm round glass cover slide to validate the effect of nitric oxide (NO) on the protein secretion from the SC and the function of permeability barrier (tight junction) formed by the SC monolayer. The results were as follows.1. The amount of NO produced spontaneously by SC in culture was relatively minute.2. The NO concentration in the conditioned media was not significantly increased by the addition of LPS (1 μg/ml) to the basal reservoir of the culture system.3. The electrical resistance (ER) of SC monolayer was not influenced by the addition of the exogenous NO that released from a NO donor (NOC18).4. The amount of the total transferrin (TF) secreted by SC was not significantly different statistically among the control and the NOC18 treated epithelial sheets. However, the polarity of the TF secretion by SC was significantly lower for the NO-do … More nor contained cultures. The secretion ratio (apical/basal=1.86 ± 0.22) of TF during 2-day collection period was significantly declined to 1.26 ± 0.04 and 1.47 ± 0.08 in the culture with the exogenously added NOC18 (50 and 100 μM), respectively.5. Immunostaining for occludin and ZO-1 in untreated-SC showed continuous labeling around the cell periphery in the region of the cell-cell junctional complex, and these colocalized with TJ-associated F-actin filaments.6. However, incubation with NOC 18 (200 μM and 400 μM) caused complete disorganization of occludin staining and partial disruption of ZO-1 staining within 48 h.7. Furthermore, descending expressions of occludin, ZO-1 and cortactin were observed in a dose-dependent manner of NOC 18 in NO donor treated SC.These results suggest that NO produced excessively within testis during acute or chronic pathophysiological conditions disrupts TJ-associated proteins of SC and that NO may perturb blood-testis barrier formation and in turn the regulation of normal spermatogenesis. Less