Protective Effect of Chlorogenic Acid on Human Sperm: In Vitro Studies and Frozen–Thawed Protocol

Antioxidants

卷:10

Andrea Menchiari¹  Daniela Cerretani²  Elena Moretti³  Daria Noto³  Cinzia Signorini³  Giulia Collodel³  Laura Gambera⁴ 

[1] Department of Business and Law, University of Siena, 53100 Siena, Italy;

[2] Department of Medicine, Surgery and Neurosciences, University of Siena, 53100 Siena, Italy;

[3] Department of Molecular and Developmental Medicine, University of Siena, 53100 Siena, Italy;

[4] Fertility Center, AGI Medica, 53100 Siena, Italy;

关键词: AMPK;    chlorogenic acid;    cryopreservation;    DNA integrity;    human sperm;    isoprostanes;   

DOI  :  10.3390/antiox10050744

来源: DOAJ

【 摘 要 】

The study evaluated the chlorogenic acid (CGA) antioxidant potential on oxidative stress (OS) induced in vitro in human spermatozoa and during cryopreservation procedure. Swim-up selected spermatozoa were treated with 100 µM CGA, 100 µM H₂O₂ to induce lipid peroxidation (LPO), and with both compounds and the effects on mitochondrial membrane potential (MMP) by JC-1, DNA integrity by acridine orange (AO), and sperm ultrastructure by transmission electron microscopy (TEM), were evaluated. CGA antioxidant activity was assessed by measuring malondialdehyde (MDA) and F₂-isoprostanes (F₂-IsoPs) in the media. The CGA protective activity and the immunolocalization of Phospho-AMPKα (Thr172) were explored in frozen-thawed sperm. CGA was not toxic for sperm motility, DNA integrity and MMP. The increase in MDA (p < 0.05) and F₂-IsoPs (p < 0.001), DNA damage (p < 0.01) and low MMP (p < 0.01) levels after H₂O₂ treatment were reduced in presence of CGA as well as the percentage of broken plasma membranes (p < 0.01) and altered acrosomes (p < 0.01) detected by TEM. Treated frozen-thawed spermatozoa showed increased sperm motility (p < 0.01), DNA integrity (p < 0.01), MMP (p < 0.01), reduced MDA (p < 0.01) and increased sperm percentage with Phospho-AMPKα labelling in the head (p < 0.001). CGA can be used to supplement culture media during semen handling and cryopreservation where OS is exacerbated.

【 授权许可】

Unknown