This study investigated the neuroprotective effects of methyl 3,4-dihydroxybenzoate (MDHB) against t-butyl hydroperoxide (TBHP) induced oxidative damage in SH-SY5Y (human neuroblastoma cells) and the underlying mechanisms. SH-SY5Y were cultured in DMEM + 10% FBS for 24 h and pretreated with different concentrations of MDHB or N-acetyl-l-cysteine (NAC) for 4 h prior to the addition of 40 μM TBHP for 24 h. Cell viability was analyzed using the methylthiazolyltetrazolium (MTT) and lactate dehydrogenase (LDH) assays. An annexin V-FITC assay was used to detect cell apoptosis rates. The 2′,7′-dichlorofluorescin diacetate (DCFH-DA) assay was used to determine intracellular ROS levels. The activities of antioxidative enzymes (GSH-Px and SOD) were measured using commercially available kits. The oxidative DNA damage marker 8-OHdG was detected using ELISA. Western blotting was used to determine the expression of Bcl-2, Bax, caspase 3, p-Akt and Akt proteins in treated SH-SY5Y cells. Our results showed that MDHB is an effective neuroprotective compound that can mitigate oxidative stress and inhibit apoptosis in SH-SY5Y cells.

This study investigated the neuroprotective effects of methyl 3,4-dihydroxybenzoate (MDHB) against t-butylhydroperoxide(TBHP) induced oxidative damage in SH-SY5Y (human neuroblastoma cells) and the underlying mechanisms. SH-SY5Y were cultured in DMEM+10% FBS for 24 hours and pretreated with different concentrations of MDHB or N-acetyl-L-cysteine (NAC) for 4 hours prior to the addition of 40 μM TBHP for 24 hours. Cell viability was analyzed using the methyl thiazolyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays. An annexin V-FITC assay was used to detect cell apoptosis rate. The 2',7'-dichlorofluorescin diacetate (DCFH-DA) assay was used to determine intracellular ROS levels. The activities of antioxidative enzymes (GSH-Px and SOD) were measured using commercially available kits. The oxidative DNA damage marker 8-OHdG was detected using ELISA. Western blotting was used to determine the expression of Bcl-2, Bax, caspase 3, p-Akt and Akt proteins in treated SH-SY5Y cells. Our results showed that MDHB is an effective neuroprotective compound that can mitigate oxidative stress and inhibit apoptosis in SH-SY5Y cells

Apoptosis plays a pivotal role in neuronal death following ischemic brain injury. Berberine (BBR), a natural isoquinoline alkaloid extracted from Coptis chinensis, exhibits multiple biological activities and has shown neuroprotective effects in experimental ischemia models. However, its direct protective effects on neurons and the underlying apoptosis-related mechanisms remain to be fully clarified. A transient forebrain ischemia model was established in mice to evaluate the neuroprotective effects of BBR in vivo. Cognitive function was assessed using the Y-maze test, and hippocampal neuronal injury was examined by H&E staining. In vitro, primary hippocampal neurons were subjected to oxygen-glucose deprivation (OGD) to mimic ischemic injury. Neuronal viability, mitochondrial function, intracellular calcium levels, apoptosis, and related signaling pathways were analyzed using CCK-8, LDH, JC-1, Fluo-4 AM, Hoechst staining, caspase-3 activity assay, and Western blotting. BBR significantly reduced hippocampal neuronal loss and improved cognitive deficits in mice after transient forebrain ischemia. In primary hippocampal neurons exposed to OGD, BBR markedly enhanced cell viability, alleviated mitochondrial dysfunction, attenuated intracellular calcium overload, and suppressed neuronal apoptosis. Mechanistically, BBR may increase the expression of phospho-Akt and Bcl2 while reducing the expression of Bax and the activity of cleaved caspase-3. These findings demonstrate that berberine exerts significant neuroprotective effects against ischemia-induced neuronal injury by targeting apoptosis-related signaling pathways, particularly the Akt/caspase-3 axis, and support its potential as a therapeutic candidate for ischemic stroke.