Bosa Luigia, Mattioli Simona, Stabile Anna Maria, Bartolini Desirée, Tognoloni Alessia, Pistilli Alessandra, Ruggirello Mariangela, Rende Mario, Gimeno-Martos Silvia, Jordán-Rodríguez Daniela, Arias-Álvarez Maria, García Rebollar Pilar, García-García Rosa M, Castellini Cesare
Department of Agricultural, Food and Environmental Sciences, University of Perugia, 06121 Perugia, Italy.
Department of Medicine and Surgery, University of Perugia, 06129 Perugia, Italy.
Biology (Basel). 2025 Aug 1;14(8):974. doi: 10.3390/biology14080974.
The aim of this study was to analyze how recombinant rabbit NGF (Nerve Growth Factor) encapsulated in chitosan (rrβNGFch) affects sperm viability, motility, capacitation, acrosome reaction (AR), kinetic traits, and apoptosis after 30 min and 2 h of storage. Specific intracellular signaling pathways associated with either cell survival, such as protein kinase B (AKT) and extracellular signal-regulated kinases 1/2 (ERK1/2), or programmed cell death, such as c-Jun N-terminal kinase (JNK), were also analyzed. The results confirmed the effect of rrβNGFch on capacitation and AR, whereas a longer storage time (2 h) decreased all qualitative sperm traits. AKT and JNK did not show treatment-dependent activation and lacked a correlation with functional traits, as shown by ERK1/2. These findings suggest that rrβNGFch may promote the functional activation of sperm cells, particularly during early incubation. The increase in capacitation and AR was not linked to significant changes in pathways related to cell survival or death, indicating a specific action of the treatment. In contrast, prolonged storage negatively affected all sperm parameters. ERK1/2 activation correlated with capacitation, AR, and apoptosis, supporting its role as an NGF downstream mediator. Further studies should analyze other molecular mechanisms of sperm and the potential applications of NGF in assisted reproduction.
本研究的目的是分析壳聚糖包裹的重组兔神经生长因子(rrβNGFch)在储存30分钟和2小时后如何影响精子活力、运动能力、获能、顶体反应(AR)、动力学特征和凋亡。还分析了与细胞存活相关的特定细胞内信号通路,如蛋白激酶B(AKT)和细胞外信号调节激酶1/2(ERK1/2),以及与程序性细胞死亡相关的通路,如c-Jun氨基末端激酶(JNK)。结果证实了rrβNGFch对获能和AR的影响,而较长的储存时间(2小时)会降低所有精子的质量性状。AKT和JNK未显示出依赖于处理的激活,且与功能性状缺乏相关性,如ERK1/2所示。这些发现表明,rrβNGFch可能促进精子细胞的功能激活,尤其是在早期孵育期间。获能和AR的增加与细胞存活或死亡相关通路的显著变化无关,表明该处理具有特定作用。相比之下,长时间储存会对所有精子参数产生负面影响。ERK1/2激活与获能、AR和凋亡相关,支持其作为NGF下游介质的作用。进一步的研究应分析精子的其他分子机制以及NGF在辅助生殖中的潜在应用。