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Effect of red propolis extract isolated or encapsulated in nanoparticles on the in vitro culture of sheep preantral follicle: Impacts on antrum formation, mitochondrial activity and glutathione levels
Author(s) -
Nascimento Tiago S.,
Silva Isabelle S. M.,
Alves Maria Cecília M. A.,
Gouveia Bruna B.,
Barbosa Lara Mariane R.,
Macedo Taís J. S.,
Santos Jamile M. S.,
Monte Alane P. O.,
Matos Maria Helena T.,
Padilha Francine F.,
LimaVerde Isabel B.
Publication year - 2019
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.13347
Subject(s) - glutathione , chemistry , high performance liquid chromatography , oxidative stress , reactive oxygen species , in vitro , andrology , biochemistry , chromatography , medicine , enzyme
Contents This study aimed to characterize the hydroethanolic extract of red propolis (HERP) and nanoparticles containing HERP for using as an additive in the culture medium of isolated ovine preantral follicles. HERP was characterized by high‐performance liquid chromatography (HPLC) and determination of flavonoid content, and the nanoparticles by the mean particle diameter, polydispersity index (PI) and encapsulation efficiency (EE). The effect of HERP (10 and 20 ηg/ml—HERP10 and HERP20 groups) and nanoparticles (NP10 and NP20 groups) on isolated secondary follicles cultured in vitro for 12 days was observed by morphological evaluation, oxidative stress markers (reactive oxygen species—ROS and glutathione—GSH) and active mitochondria. HPLC showed formononetin as the major compound in the HERP (63.92 ± 0.21 μg/mL). The content of flavonoids ranged from 2.14% to 11.00%. The nanoparticles showed mean diameter of 287.5 ± 3.9 and 479 ± 18.1 ηm; PI of 0.117 ± 0.018 and 0.316 ± 0.039; and EE of 67.15% and 41%, respectively, for the NP10 and NP20 groups. After 12 days of culture, HERP20 and NP20 increased ( p < 0.05) the percentage of normal follicles compared to NP10. HERP20 showed significantly higher percentages of antrum formation than control medium (MEM) and NP10 ( p < 0.05). HERP20 also showed an increase ( p < 0.05) in mitochondrial activity compared to the other treatments, except NP20 ( p > 0.05), and increased GSH levels ( p < 0.05) compared to MEM and HERP10. In conclusion, use of HERP (20 ηg/ml) on in vitro culture of isolated ovine preantral follicles can increase antrum formation, mitochondrial activity and GSH levels.