目的分离培养大鼠前列腺毛细血管内皮细胞,体外研究大鼠前列腺毛细血管内皮细胞的屏障功能。方法采用细胞生长特性选择法分离培养大鼠前列腺毛细血管内皮细胞,用第Ⅷ因子相关抗原抗体对分离培养的前列腺毛细血管内皮细胞进行免疫学鉴定。免疫组化观察毛细血管内皮细胞之间紧密连接蛋白claudin-1的表达,采用跨膜电阻测量仪检测毛细血管内皮细胞的跨膜电阻,用酚红渗漏实验检测其通透性,以大鼠肺毛细血管内皮细胞、脑毛细血管内皮细胞作为阴性与阳性对照。结果成功分离大鼠前列腺毛细血管内皮细胞,体外培养可以形成紧密的单层细胞结构,呈铺路石样生长。第Ⅷ因子相关抗原抗体的免疫荧光结果显示:分离培养的大鼠前列腺毛细血管内皮细胞胞浆表达绿色荧光,提示分离的细胞为毛细血管内皮细胞;免疫组化结果显示紧密连接蛋白claudin-1在相邻的毛细血管内皮细胞之间表达;体外培养的大鼠前列腺毛细血管内皮细胞,阴性对照组肺毛细血管内皮细胞,阳性对照组脑毛细血管内皮细胞的跨膜电阻峰值分别可以达到(142.2±3.1)、(43.3±3.5)、(248.2±6.2)Ω/cm2,并稳定保持2~3 d,三者比较差异有统计学意义(P<0.05)。酚红渗漏实验结果显示前列腺毛细血管内皮细胞、阳性对照组脑毛细血管内皮细胞随着细胞单层跨膜电阻的增加,基底侧的酚红浓度减低,阴性对照组肺毛细血管内皮细胞酚红渗透量随跨膜电阻的改变变化趋势不明显。结论大鼠前列腺毛细血管内皮细胞与脑毛细血管内皮细胞等屏障内皮相比拥有相似的结构功能,体外培养的大鼠前列腺毛细血管内皮细胞具有屏障特性,可以作为血前列腺屏障研究的体外模型。 Objective To isolate and culture rat prostatic capillary endothelial cells (ECs) and study the barrier function of rat prostatic capillary endothelial cells (ECs) in vitro. Methods The rat prostatic capillary endothelial cells were isolated and cultured by the method of cell growth selection. Immunohistochemistry was used to identify the prostatic capillary endothelial cells isolated by the factor Ⅷ antibody. Immunohistochemistry was used to observe the expression of tight junction protein claudin-1 between capillary endothelial cells. The transmembrane resistance of capillary endothelial cells was measured by transmembrane resistance meter. The permeability of capillary endothelial cells was detected by phenol red leakage assay. Pulmonary capillary endothelial cells, brain capillary endothelial cells as a negative and positive control. Results The rat prostatic capillary endothelial cells were successfully isolated and cultured in vitro to form a compact monolayer cell structure with paving stone-like growth. The results of immunofluorescence assay showed that the cytoplasm of isolated rat prostatic capillary endothelial cells expressed green fluorescence, suggesting that the isolated cells were capillary endothelial cells. The results of immunohistochemistry showed that claudin-1 Adjacent capillary endothelial cells; the peak value of transmembrane resistance in rat prostatic capillary endothelial cells cultured in vitro, pulmonary capillary endothelial cells in the negative control group and brain capillary endothelial cells in the positive control group were respectively (142.2 ± 3.1), (43.3 ± 3.5) and (248.2 ± 6.2) Ω / cm2, respectively. The difference was statistically significant (P <0.05). Phenol red leakage experimental results showed that with the increase of cell monolayer transmembrane resistance, the concentration of phenol red in the basal side decreased in the prostatic capillary endothelial cells and in the positive control group, while in the negative control group, the level of phenol red in the pulmonary capillary endothelial cells The change of permeability with the change of membrane resistance is not obvious. Conclusions Rat prostatic capillary endothelial cells have similar structural functions as barrier endothelium such as brain capillary endothelial cells. Rat prostatic capillary endothelial cells cultured in vitro have barrier properties that can be used as an in vitro model for studying the prostate gland barrier.