The study aimed to analyze changes in biomolecular composition of granulosa and theca interna cells of porcine ovarian follicles following in vitro treatment of vitamin D₃ and insulin alone or in combination. Medium antral follicles (n = 4/each group) were cultured alone (C; control) or in the presence of 1α,25(OH)₂D₃ (VD; 100 ng/mL) and insulin (I; 10 ng/mL) separately or in combination, VD and I (VD+I). Then paraplast-embedded ovarian follicles were used for Fourier Transform Infrared (FTIR) spectroscopy and respective histological stainings. FTIR analysis revealed changes in the content of fibrous proteins (mainly collagens) within theca interna following vitamin D₃ and insulin co-administration that was verified by Masson’s trichrome staining. Treatment-dependent differences were also observed in the secondary structure of proteins, indicating enhanced conversion to α-helices in response to vitamin D₃ and insulin action/interaction in both follicular compartments. In the granulosa and theca interna layers, tendency to lower DNA content in the VD+I group was noted and confirmed by Fulgen’s staining. Finally, altered monosaccharides production in both follicular layers was found. Based on FTIR results, it is possible to attribute the observed alterations to biological processes that could be regulated by vitamin D₃ and insulin in the porcine ovarian follicles.

该研究旨在分析单独或联合使用维生素 D _{3和胰岛素}体外处理后猪卵泡颗粒和卵泡膜内细胞生物分子组成的变化。中型窦卵泡（n = 4/每组）单独培养（C；对照）或在 1α,25(OH) ₂ D ₃（VD；100 ng/mL）和胰岛素（I；10 ng/mL）存在下培养) 单独或组合，VD 和 I (VD+I)。然后将包埋的卵泡用于傅里叶变换红外（FTIR）光谱和各自的组织学染色。_{FTIR 分析揭示了维生素 D 3}和胰岛素共同给药后内膜内纤维蛋白（主要是胶原蛋白）含量的变化，这一点通过 Masson 三色染色得到了验证。在蛋白质二级结构中也观察到了治疗依赖性差异，表明两个卵泡室中响应维生素 D _{3和胰岛素作用/相互作用而增强的 α-螺旋转化。}在颗粒层和卵泡膜内层中，VD+I组中DNA含量降低的趋势被注意到并通过Fulgen染色证实。最后，发现两个毛囊层中单糖的产生发生了变化。根据 FTIR 结果，可以将观察到的变化归因于猪卵巢卵泡中维生素 D _{3和胰岛素调节的生物过程。}