C caspase 8

The prostates were immediately fixed in 4% paraformaldehyde at 4 °C for 24 h and then in 50% ethanol before processing for histological studies. Serial sections (5 μm thickness; 1 section per slide) were performed for histological and immunohistochemistry (IHC) analyses. For histology, sections were subjected to hematoxylin and eosin (H&E) staining and were examined using a light microscope (Nikon Eclipse E100, Tokyo, Japan). Fields were selected following systematic random sampling scheme, and the epithelial thickness was measured using ImageJ software. For IHC studies, after deparaffinization and dehydration, sections were washed with 3% H₂O₂ for 10 min at room temperature, and non-specific binding of immunoglobulins was blocked with 2% normal goat serum (diluted 1:10 in PBS) for 30 min at room temperature. Sections were boiled for 30 min in critic acid (pH 6.0) for antigen retrieval. After blocking, sections were incubated overnight with primary antibodies, as follows: Ki-67, C-Caspase-3, C-Caspase-8, C-Caspase-9 (Proteintech Group, Inc). Images were acquired using a light microscope (Nikon Eclipse E100, Tokyo, Japan).

Testosterone propionate was purchased from Ningbo Second Hormone Factory and finasteride was purchased from CONBA Co. LTD, Zhejiang. The Rat dihydrotestosterone (DHT), 5α-reductase, IL-1β, IL-6 and TNF-α ELISA kits were provided by Jiangsu Enzyme Standard Biotechnology Co., Ltd (Jiangsu, China). Protein Extraction Solution was purchased from Dingguo Changsheng Biotechnology Co., Ltd (Beijing). Antibodies for Ki-67, C-Caspase-3, C-Caspase-8 and C-Caspase-9 were purchased from Proteintech Group, Inc (American).