For viral infection in vivo, 6 days prior to HIBD, a vector containing TIGAR short hairpin RNA (LV-sh_TIGAR; 1×109 transduction units/mL; Genechem, Shanghai, China) or GSDMD short hairpin RNA (LV-sh_GSDMD; 1 × 109 transduction U/mL; Genechem) was infused into the left lateral ventricle and striatum (2 μL/injection site) (Li et al., 2014) of the neonatal rats. Nicotinamide adenine dinucleotide phosphate (NADPH; 2.5 mg/kg; Beyotime, Nantong, China) was injected into the left lateral ventricle of the rat brain before HIBD modeling.
For viral infection in vitro, the HAPI rat microglial cells were treated with medium containing LV-sh_TIGAR (MOI = 100), LV-sh_GSDMD (MOI = 100), or LV-sh-scramble (normal control; MOI = 100; Genechem), and the medium was replaced with regular medium after 12 hours of viral treatment. The cells were cultured for 3 days and then subjected to OGD/reoxygenation (OGD/R). NADPH (10 μM; Beyotime) was added before OGD and was present in the regular medium used for reoxygenation at 24 hours.
For viral infection in vitro, the HAPI rat microglial cells were treated with medium containing LV-sh_TIGAR (MOI = 100), LV-sh_GSDMD (MOI = 100), or LV-sh-scramble (normal control; MOI = 100; Genechem), and the medium was replaced with regular medium after 12 hours of viral treatment. The cells were cultured for 3 days and then subjected to OGD/reoxygenation (OGD/R). NADPH (10 μM; Beyotime) was added before OGD and was present in the regular medium used for reoxygenation at 24 hours.