Jin Yin-Shan, Cui Yuan-Qing, Xu Yan-Ping, Chen Jie, Zhang Xue-Bao, Wang Xiong
Department of Reproductive Medicine, Yantai Yuhuangding Hospital, Yantai, 264000, People's Republic of China.
J Inflamm Res. 2024 Oct 13;17:7261-7274. doi: 10.2147/JIR.S476276. eCollection 2024.
Varicocele is a dilatation of the internal spermatic vein and it is generally recognized as one cause of male infertility. This study aimed to analyze the roles of activating transcription factor 6 (ATF-6) in experimental varicocele-induced epididymal epithelial cells.
Experimental left varicocele was established in rats through partial left renal vein ligation. At 8 weeks after surgery, the left epididymal damage was observed using H&E and TUNEL staining. The expressions of neutral α-glucosidase (NAG), ATF-6, tumor necrosis factor (TNF)-α, and phospho-nuclear factor (p-NF)-κB p65 (S536) in the left epididymis were measured by immunohistochemistry. ATF-6 silence in rat epididymal epithelial cells was established by ATF-6 siRNA transfection. The cells were treated with hypoxia for 24 h, and cell viability was measured by CCK-8, levels of NAG, TNF-α, and interleukin (IL)-8 in cells were measured by ELISA, levels of p-NF-κB p65 (S536)/NF-κB p65 protein in cells were measured by Western blotting.
The results showed that the experimental left varicocele induced hypertrophy and apoptosis of epididymal epithelial cells (p<0.05), and decreased the expressions of NAG in the epididymal epithelial cells compared with the sham-operated control rats (p<0.01). Meanwhile, the expressions of ATF-6, TNF-α, and p-NF-κB p65 (S536) were increased in the epididymal epithelial cells after the experimental left varicocele compared with the sham-operated control rats (p<0.05). In the hypoxia-treated cells, ATF-6 silence increased the cell viability and decreased the levels of TNF-α, IL-8, and p-NF-κB p65 (S536) compared with the control cells (p<0.05).
The ATF-6 pathway was activated in a rat's left varicocele-induced epididymal damage. Inhibition of the ATF-6 pathway might be a possible novel therapeutic approach for left varicocele-induced epididymal damage.
精索静脉曲张是精索内静脉的扩张，通常被认为是男性不育的原因之一。本研究旨在分析激活转录因子6（ATF-6）在实验性精索静脉曲张诱导的附睾上皮细胞中的作用。
通过部分结扎左肾静脉在大鼠中建立实验性左侧精索静脉曲张。术后8周，使用苏木精-伊红（H&E）和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色观察左侧附睾损伤情况。采用免疫组织化学法检测左侧附睾中中性α-葡萄糖苷酶（NAG）、ATF-6、肿瘤坏死因子（TNF）-α和磷酸化核因子（p-NF）-κB p65（S536）的表达。通过ATF-6小干扰RNA（siRNA）转染建立大鼠附睾上皮细胞中ATF-6沉默。将细胞缺氧处理24小时，采用细胞计数试剂盒-8（CCK-8）检测细胞活力，采用酶联免疫吸附测定（ELISA）法检测细胞中NAG、TNF-α和白细胞介素（IL）-8水平，采用蛋白质免疫印迹法检测细胞中p-NF-κB p65（S536）/NF-κB p65蛋白水平。
结果显示，与假手术对照组大鼠相比，实验性左侧精索静脉曲张诱导附睾上皮细胞肥大和凋亡（p<0.05），并降低附睾上皮细胞中NAG的表达（p<0.01）。同时，与假手术对照组大鼠相比，实验性左侧精索静脉曲张后附睾上皮细胞中ATF-6、TNF-α和p-NF-κB p65（S536）的表达增加（p<0.05）。在缺氧处理的细胞中，与对照细胞相比，ATF-6沉默增加了细胞活力并降低了TNF-α、IL-8和p-NF-κB p65（S536）水平（p<0.05）。
ATF-6通路在大鼠左侧精索静脉曲张诱导的附睾损伤中被激活。抑制ATF-6通路可能是治疗左侧精索静脉曲张诱导的附睾损伤的一种新的潜在治疗方法。
