The variable expression of lectin-like oxidized low-density lipoprotein receptor (LOX-1) and signs of autophagy and apoptosis in freshly harvested human granulosa cells depend on gonadotropin dose, age, and body weight

To extend our recent observations on lectin-like oxidized low-density lipoprotein receptor (LOX-1) expression in human granulosa cell cultures with freshly harvested granulosa cells. Design: Clinical research. Setting: Institute of Anatomy and Clinic for Reproductive Medicine. Patient(s): Women undergoing IVF therapy were classified by total FSH dose, age, and body mass index. Main outcome measure(s): Purified granulosa cells were studied by Western blot and morphology for the presence of LOX-1, microtubule-associated light-chain protein 3 (LC3) and autophagosomes, which are both autophagic markers, cleaved caspase-3 for apoptosis, and apoptosis-inducing factor (AIF) for caspase-independent apoptosis. Intervention(s): None.

Active LOX-1 was found in all samples, being at its maximum in the younger obese group with a total FSH dose <2,000 IU. The LC3 II/LC3 I ratio, indicative of reparative autophagy, was at its maximum in younger normal-weight patients and increased under total FSH dose >2,000 IU. Autophagosomes in ultrathin sections were indicative of reparative autophagy. Cleaved caspase-3 was absent in all groups. The apoptotic AIF form was up-regulated in older patients. Unpurified granulosa cells consisted of approximately 20% dead cells in the younger normal-weight group compared with up to 50% in the older obese group.