Estrogen

Individual blood samples were collected from the aortas of rats in all groups that had been fasted one day before. Serum samples were separated from whole blood by using a blood collection tube (Vacutainer; BD Biosciences, NJ, USA). All blood serum was stored at −80 °C, and anti-Müllerian hormone (AMH) (Elabscience Biotechnology, TX, USA), estrogen (BioVision, Milpitas, MA, USA) and lactate dehydrogenase (LDH) (Abcam, Waltham, MA, USA) activity in serum was analyzed by ELISA kits following the manufacturer’s instructions.

All blood samples were collected from the aortas of rats in the normal, NTx and Tx groups. Individual serum samples were separated from whole blood by using a blood collection tube (Vacutainer; BD Biosciences, San Jose, CA, USA). All blood serum was stored at − 80 °C, and estrogen (Biovision, Milpitas, CA, USA), AMH (Elabscience Biotechnology, MA, USA), FSH (Abnova, Taipei, Taiwan), and testosterone (Mybiosource, San Diego, CA, USA) activity in serum was analyzed by ELISA kits following the manufacturer’s instructions. In brief, an equal volume of sample was added to the specific antibody-coated plates. Next, specific horseradish peroxidase (HRP)-conjugates were added to each well and incubated at 37 °C. After the substrates had been added and incubated in the dark for substrate development, the antibody activity was analyzed by using a microplate reader (BioTek, Winooski, VT, USA).