The Effect of Ephedrine on Neuronal Apoptosis in Cerebral Ischemia-Reperfusion Model Mice by Regulating the Keap1/Nrf2 Signaling Pathway

This study aimed to investigate the effect of EPH (ephedrine) on neuronal apoptosis in mice with cerebral I/R (ischemia-reperfusion) model by regulating the Keap1/Nrf2 signaling pathway. The mouse model of cerebral I/R was established by occlusion of the MCAO (middle cerebral artery) and grouped into model group, L-EPH group, H-EPH group, and H-EPH+ML385 group, with 12 mice in each group. Additionally, 12 healthy mice were included as the sham surgery group. After the administration, the neurological function of mice in each group was evaluated. TCC was applied to measure the area of cerebral infarction. HE staining was applied to evaluate pathological damage in the hippocampus. Nissl staining was applied to observe neuronal damage in the hippocampus. TUNEL staining was applied to detect neuronal apoptosis in brain tissue. The reagent kit was used to determine the content of MDA (malondialdehyde) and the activity of superoxide SOD (superoxide dismutase). qRT-PCR was applied to detect the expression of Keap1 and Nrf2 mRNA in brain tissue. Western blot was applied to detect the expression of Keap1, Nrf2 and apoptosis-related proteins Caspase-3 and Bcl-2 in brain tissue; and immunofluorescence staining was used to locate Nrf2 protein. The results showed that compared with the sham surgery group, the model group showed neuronal damage in I/R mice, increased neurological deficit score, increased cerebral infarction area, decreased Nrf2 mRNA and protein expressions levels, decreased Nissl body number, increased neuronal apoptosis rate, increased Keap1 mRNA and protein expressions levels, increased Caspase-3 protein expression levels, and decreased Bcl-2 protein expression levels (P<0.05). The I/R mice in the L-EPH and H-EPH groups had alleviated neurological damage, decreased neurological deficit score, decreased cerebral infarction area, decreased Nrf2 mRNA and protein expressions levels, increased Nissl body number, decreased neuronal apoptosis rate, increased Keap1 mRNA and protein expressions levels, decreased Caspase-3 protein expression levels, and increased Bcl-2 protein expression levels (P<0.05), the H-EPH group was superior to the L-EPH group (P<0.05). The addition of Nrf2 inhibitor ML385 inhibited Nrf2 expression and reversed the healing effect of EPH on I/R mice (P<0.05). In summary, EPH can inhibit neuronal apoptosis in brain I/R mice, which may be related to regulating the Keap1/Nrf2 pathway.

CSTR: 32200.14.cjcb.2025.01.0006