Abstract：
BACKGROUND: The presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and its receptor in various testicular cells and spermatozoa suggests a potential role in enhancing spermatogonial and postmeiotic cell development. Moreover, GM-CSF activates the pivotal pathways implicated in sperm motility regulation and glucose metabolism. However, the impact of GM-CSF on testicular biopsies from patients with obstructive azoospermia (OA) remains unexplored. Therefore, this study aimed to investigate the in vitro effects of GM-CSF on the expression of genes related to glucose transporters and signaling pathways, sperm motility, and viability in testicular biopsies.
RESULTS: Following testicular sperm extraction from 20 patients diagnosed with OA, each sample was divided into two parts: the experimental samples were incubated with medium containing 2 ng/ml GM-CSF at 37 °C for 60 min, and the control samples were incubated with medium without GM-CSF. Subsequently, the oocytes retrieved from the partner were injected with sperm from the treatment and control groups. The sperm parameters (motility and viability), the expression levels of sperm motility-related genes (PIK3R1, PIK3CA, and AKT1), and the expression levels of sperm energy metabolism-related genes (GLUT1, GLUT3, and GLUT14) were assessed. Furthermore, the fertilization and day 3 embryo development rate and embryo quality were evaluated. Compared with those in the nontreated group, the motility parameters and the mRNA expression levels of PIK3R1, AKT1, and GLUT3 in testicular sperm supplemented with GM-CSF were significantly greater (p < 0.05). However, no significant differences in the mRNA expression of PIK3CA, GLUT1, or GLUT14 were detected. According to the ICSI results, compared with the control group, the GM-CSF treatment group exhibited significantly greater fertilization rates (p = 0.027), Day 3 embryo development rate (p = 0.001), and proportions of good-quality embryos (p = 0.002).
CONCLUSIONS: GM-CSF increased the expression of genes related to motility and the energy metabolism pathway and effectively promoted the motility of testis-extracted spermatozoa, consequently yielding positive clinical outcomes.
RESULTS: Following testicular sperm extraction from 20 patients diagnosed with OA, each sample was divided into two parts: the experimental samples were incubated with medium containing 2 ng/ml GM-CSF at 37 °C for 60 min, and the control samples were incubated with medium without GM-CSF. Subsequently, the oocytes retrieved from the partner were injected with sperm from the treatment and control groups. The sperm parameters (motility and viability), the expression levels of sperm motility-related genes (PIK3R1, PIK3CA, and AKT1), and the expression levels of sperm energy metabolism-related genes (GLUT1, GLUT3, and GLUT14) were assessed. Furthermore, the fertilization and day 3 embryo development rate and embryo quality were evaluated. Compared with those in the nontreated group, the motility parameters and the mRNA expression levels of PIK3R1, AKT1, and GLUT3 in testicular sperm supplemented with GM-CSF were significantly greater (p < 0.05). However, no significant differences in the mRNA expression of PIK3CA, GLUT1, or GLUT14 were detected. According to the ICSI results, compared with the control group, the GM-CSF treatment group exhibited significantly greater fertilization rates (p = 0.027), Day 3 embryo development rate (p = 0.001), and proportions of good-quality embryos (p = 0.002).
CONCLUSIONS: GM-CSF increased the expression of genes related to motility and the energy metabolism pathway and effectively promoted the motility of testis-extracted spermatozoa, consequently yielding positive clinical outcomes.
摘要：
背景：粒细胞-巨噬细胞集落刺激因子（GM-CSF）及其受体在各种睾丸细胞和精子中的存在表明在增强精原和减数分裂后细胞发育中具有潜在作用。此外，GM-CSF激活与精子运动调节和葡萄糖代谢有关的关键途径。然而,GM-CSF对梗阻性无精子症(OA)患者睾丸活检的影响仍未被研究.因此,本研究旨在探讨GM-CSF对葡萄糖转运体相关基因表达及信号通路的影响,精子运动性,和睾丸活检的活力。
结果：从20例诊断为OA的患者中提取睾丸精子后，每个样品分为两部分：将实验样品与含有2ng/mlGM-CSF的培养基在37°C孵育60分钟，和对照样品用不含GM-CSF的培养基孵育。随后，从伴侣身上取出的卵母细胞被注射治疗组和对照组的精子。精子参数（运动性和活力），精子运动相关基因的表达水平(PIK3R1，PIK3CA，和AKT1），并对精子能量代谢相关基因(GLUT1、GLUT3和GLUT14)的表达水平进行评估。此外，评价受精率和第3天胚胎发育率和胚胎质量。与未治疗组相比，补充GM-CSF的睾丸精子的运动参数和PIK3R1，AKT1和GLUT3的mRNA表达水平显着增加（p<0.05）。然而,PIK3CA的mRNA表达无显著差异,检测到GLUT1或GLUT14。根据ICSI的结果,与对照组相比，GM-CSF治疗组表现出明显更大的受精率（p=0.027），第3天胚胎发育率（p=0.001），和优质胚胎的比例（p=0.002）。
结论：GM-CSF增加了运动性和能量代谢途径相关基因的表达，有效促进了睾丸摘除精子的运动性。因此产生积极的临床结果。
结果：从20例诊断为OA的患者中提取睾丸精子后，每个样品分为两部分：将实验样品与含有2ng/mlGM-CSF的培养基在37°C孵育60分钟，和对照样品用不含GM-CSF的培养基孵育。随后，从伴侣身上取出的卵母细胞被注射治疗组和对照组的精子。精子参数（运动性和活力），精子运动相关基因的表达水平(PIK3R1，PIK3CA，和AKT1），并对精子能量代谢相关基因(GLUT1、GLUT3和GLUT14)的表达水平进行评估。此外，评价受精率和第3天胚胎发育率和胚胎质量。与未治疗组相比，补充GM-CSF的睾丸精子的运动参数和PIK3R1，AKT1和GLUT3的mRNA表达水平显着增加（p<0.05）。然而,PIK3CA的mRNA表达无显著差异,检测到GLUT1或GLUT14。根据ICSI的结果,与对照组相比，GM-CSF治疗组表现出明显更大的受精率（p=0.027），第3天胚胎发育率（p=0.001），和优质胚胎的比例（p=0.002）。
结论：GM-CSF增加了运动性和能量代谢途径相关基因的表达，有效促进了睾丸摘除精子的运动性。因此产生积极的临床结果。
