Anti p38

Berberine (purity: ≥98%) and cisplatin (purity: ≥99%) were purchased from Sigma Aldrich (St. Louis, MO, USA). Anti-MMP-2, anti-MMP-9, anti-Bcl-2, anti-Bax, anti-CyclinD1, anti-CDK4, anti-JNK, anti-phospho-JNK, anti-ERK, anti-phospho-ERK, anti-P38, and anti-phospho-P38 primary antibodies were purchased from Bioss (Beijing, China). Goat antimouse IgG and goat antirabbit IgG secondary antibodies were purchased from Life Science (Santa Cruz, CA, USA).

28 days following SCI, euthanized rats under isoflurane anesthesia. Samples were then collected and fixed in 4% paraformaldehyde. A 0.5 cm long sample tissue was homogenized in 300 μL of lysate-containing protease inhibitors. Following lysate preparation, the electrophoresed tissue lysate was transferred to polyvinylidene difluoride membranes. Next, the membranes were blocked with 5% skimmed milk and incubated in an incubator for 60 min. The membranes were further blocked for two hours and then incubated with primary antibodies, consisting of anti-JNK (1:1000; Proteintech), anti-p-JNK (1:1000; Proteintech), anti-p-p38 (1:500; Bioss), anti-p38 (1:1000; Bioss), and GAPDH (1:2000, UtiBody). They were then incubated with secondary antibodies (1:3000; Bioss) in an incubator for 1 h. ImageJ was used to quantify grey bands.

D-gal (≥99%) was purchased from Sigma Aldrich (St. Louis, MO, USA). Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and 8-hydroxy deoxyguanosine (8-OHdG) commercial kits were purchased from Nanjing Jiancheng Institute of Biotechnology (Nanjing, China). Testosterone, cortisol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-β1 (TNF-β1), and tumor necrosis factor-α (TNF-α) enzyme-linked immunosorbent assay (ELISA) kits were obtained from Nanjing SenBeiJia Biological Technology Co., Ltd (Nanjing, China). Anti-p-p38, anti-p-ERK1/2, anti-p-JNK, anti-p38, anti-ERK1/2 and anti-JNK antibodies were purchased from Bioss (Beijing, China). The primary antibody for GAPDH was from Biogot Technology Co., Ltd. (Nanjing, Jiangsu, China). Anti-rabbit secondary antibody was obtained from Cell Signaling Technology (Danvers, MA, USA). All other reagents used in the experiments were of analytical grade.