The effects of salvianolic acid B （SalB） on the mitochondrial membrane potential （MMP）, calcium, and apoptosis of neurons with cerebral ischemia in mice were investigated using an acute cerebral ischemia model established by ligating the bilateral common carotid arteries in mice. The MMP, the intracellular cal- cium concentration, and the apoptosis rate of cortical neurons were measured at 6 min, 12 rain, 18 rain, 24 min, and 30 min after cerebral ischemia by a flow cytometer. The experiments show that SalB increases the MMP and reduces the intracellular calcium and the apoptosis rate at different stages of the cerebral ischemia in mice. The results show that the protective mechanism of SalB on cerebral ischemia enhances the MMP and maintains intracellular calcium homeostasis.
Background Cerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key issue to be solved. This research aimed to investigate the neuroprotective effects of salvianolic acid B （SalB） against oxygen-glucose deprivation/reperfusion （OGD/RP） damage in primary rat cortical neurons.Methods The primary cultures of neonatal Wister rats were randomly divided into the control group, the OGD/RP group and the SalB-treatment group （10 mg/L）. The cell model was established by depriving of oxygen and glucose for 3 hours and reperfusion for 3 hours and 24 hours, respectively. The neuron viability was determined by MTT assay. The level of cellular reactive oxygen species （ROS） was detected by fluorescent labeling method and spin trapping technique respectively. The activities of neuronal Mn-superoxide dismutase （Mn-SOD）, catalase （CAT） and glutathione peroxidase （GSH-PX） were assayed by chromatometry. The mitochondria membrane potential （△ψm） was quantitatively analyzed by flow cytometry. The release rate of cytochrome c was detected by Western blotting. The neuronal ultrastructure was observed by transmission electron microscopy. Statistical significance was evaluated by analysis of variance （ANOVA）followed by Student-Newman-Keuls test.Results OGD/RP increased the level of cellular ROS, but decreased the cell viability and the activities of Mn-SOD, CAT and GSH-PX; SalB treatment significantly reduced the level of ROS （P ＜0.05）; and enhanced the cell viability （P ＜0.05）and the activities of these antioxidases （P ＜0.05）. Additionally, OGD/RP induced the fluorescence value of △ψm to diminish and the release rate of cytochrome c to rise notably; SalB markedly elevated the level of △ψm （P ＜0.01） and depressed the release rate of cytochrome c （P ＜0.05）; it also ameliorated the neuronal morphological injury.Conclusion Th
WANG YunJIANG Yu-fengHUANG Qi-fuGE Gui-lingCUI Wei
Objetive： TO investigate the neuroprotective effects and underlying mechanisms of salvianolic acid B （Sal B） extracted from Salvia miltiorrhiza on hippocampal CA1 neurons in mice with cerebral ischemia reperfusion injury. Methods： Forty male National Institute of Health （NIH） mice were randomly divided into 4 groups with 10 animals each, including the sham group, the model group, the SalB group （SalB 22.5 mg/kg） and the nimodipine （Nim） group （Nim 1 mg/kg）. A mouse model of cerebral ischemia and reperfusion injury was established by bilateral carotid artery occlusion for 30 rain followed by 24-h reperfusion. The malondialdehyde （MDA） content, the nitric oxide synthase （NOS） activity, the superoxide dismutase （SOD） activity and total anti- oxidant capability （T-AOC） of the pallium were determined by biochemistry methods. The morphologic changes and Bcl-2 and Bax protein expression in hippocampal CA1 neurons were observed by using hematoxylin- eosin staining and immunohistochemistry staining, respectively. Results： In the SalB group, the MDA content and the NOS activity of the pallium in cerebral ischemia-reperfusion mice significantly decreased and the SOD activity and the T-AOC significantly increased, as compared with the model group （P〈0.05 or P〈0.01）. The SalB treatment also rescued neuronal loss （P〈0.01） in the hippocampal CA1 region, strongly promoted Bcl-2 protein expression （P〈0.01） and inhibited Bax protein expression （P〈0.05）. Conclusions： SalB increases the level of antioxidant substances and decreases free radicals production. Moreover, it also improves Bcl-2 expression and reduces Bax expression. SalB may exert the neuroprotective effect through mitochondria-dependent pathway on hippocampal CA1 neurons in mice with cerebral ischemia and reperfusion injury and suggested that SalB represents a promising candidate for the prevention and treatment of ischemic cerebrovascular disease.
