Bms629

At 24 h and 72 h post-SAH, rats were anesthetized and transcardially perfused with 250 mL PBS, the left hemisphere was harvested immediately and homogenized in 0.9% normal saline at 200 mg/mL and centrifuged at 12,000×g for 20 min. The supernatant was collected and stored in − 80 °C until further used. The concentrations of TNF-α, interleukin (IL)-6, IL-1β, and IL-10 in brain tissue lysates were analyzed using commercial ELISA kits (BMS625, BMS630, and BMS629, Invitrogen) according to the manufacturer’s instructions. The final concentration of cytokines was interpolated from the determined standard curve of absorbance.

At 24 h after SAH, rats were deeply anesthetized and serum samples from the left hemisphere were obtained and stored at −80°C until use. The concentrations of IL-10, IL-1β, and IL-18 in brain tissue lysates were analyzed using commercial ELISA kits (BMS625, BMS630, and BMS629, Invitrogen) according to the manufacturer's instructions. The final concentration of cytokines was obtained from the determined standard curve of absorbance.

At 24 h post-SAH, rats were anesthetized and transcardially perfused with 250 mL pre-cooled PBS (0.1 M, pH 7.4). Brains were then removed and rapidly divided into the left and right hemisphere; the left hemisphere was frozen in an ultra-low temperature refrigerator, homogenized in PBS (0.1 M, pH 7.4) at 200 mg/mL, and centrifuged at 12,000 g for 15 min. The liquid supernatant was collected and stored at −80 °C until further use. The concentrations of TNF-α, IL-1β, IL-17and IL-18 were analyzed using specific ELISA kits (BMS625, BMS630, and BMS629, respectively, Invitrogen) according to the manufacturer’s instructions. The concentration of cytokines was expressed by the intensity of absorbance measured by spectrometry on a microplate reader (Varioskan Lux, Thermo Scientific). Results were expressed in picograms per milliliter (pg/ml).