EGb761 Ameliorates Neuronal Apoptosis and Promotes Angiogenesis in Experimental Intracerebral Hemorrhage via RSK1/GSK3ß Pathway.

Neuronal apoptosis after intracerebral hemorrhage (ICH) plays an essential role in neurological deterioration. Preclinical studies have shown that EGb761, an extract of Ginkgo biloba, is neuroprotective in some other neurological diseases with apoptosis. This study was conducted to investigate the potential neuroprotective effect of EGb761 on neuronal apoptosis in experimental ICH. A model of ICH was induced in C57BL/6 mice by injecting collagenase. EGb761 was administered for 21 days and neurologic behaviors were assessed at 1, 3, 7, 14, and 21 days after ICH. RNAi-mediated knockdown of p90 ribosomal S6 kinase 1 (RSK1) was used to further investigate the role of RSK1 in EGb761-induced neuroprotective effects. Neuronal death was determined by TUNEL staining. The image datasets of neurovascular networks were acquired via micro-optical sectioning tomography (MOST). The glycogen synthase kinase-3ß (GSK3ß) activity was assayed using commercial kit. Primary cultured cortical neurons were exposed to ferrous iron and treated with EGb761. Apoptotic neurons were counted by flow cytometry. RSK1, GSK3ß, phosphorylated-GSK3ß (pGSK3ß), Bcl2, Bax, cleaved-caspase3 (CC3), and VEGF were measured by Western blot. The pGSK3ß was also detected by immunofluorescence staining. We found that mice in EGb761 group performed better on rotarod test. Reduced TUNEL-positive neurons and richer microvascular networks were observed in mice treated with EGb761. EGb761 attenuates neuronal apoptosis induced by ferrous iron counted by flow cytometry in vitro. Decreased GSK3ß activity was observed in EGb761-treated mice compared with mice with ICH. EGb761 increased the expression of pGSK3ß (Ser9), RSK1 and the Bcl2/Bax ratio, and VEGF and decreased CC3 expression. In conclusion, EGb761 reduces neuronal apoptosis and promotes angiogenesis in experimental intracerebral hemorrhage via RSK1/GSK3ß pathway.

[Acanthopanax senticosus saponins ameliorates oxidative damage induced by hydrogen peroxide in neonatal rat cardiomyocytes].

OBJECTIVE: To investigate the effect of Acanthopanax senticosus saponins (ASS) on oxidative damage induced by hydrogen peroxide (H2O2 ) in cardiomyocytes. METHOD: The cardiomyocytes were induced to oxidative damaged by exposed to H2O2. We evaluated the level of oxidative injury through morphology change, the survival rate, the activity of lactate dehydrogenase (LDH) and the content of cellular malondialdehyde (MDA). The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and the content of reductive glutathione (GSH) of cardiomyocytes were also determined. RESULT: When the cardiomyocytes were exposed to H2O, (50, 100, 200 micromol x L(-1)) for deigned time, the percentage of survival cells was down significantly (P < 0.01 or P < 0.001), and the activity of LDH and the content of MDA were increased markedly (P < 0.05, P < 0.01 or P < 0.001). These results show that the cells were subjected to oxidative damage. Treatment with ASS (600 mg x L(-1)) prior to H2O2 exposure could increase cell viability significantly, lessen cardiomyocyte morphological damaged change, and decrease LDH activity (1687.40 +/- 97.51) U x mL(-1) in media and cellular MDA content (16.50 +/- 2.66) nmol x mg(-1) markedly (P < 0.01 and P < 0.05). Furthermore, the activities of SOD (89.55 +/- 6.93) U x mg(-1), GSH-Px (845.87 +/- 63.76) mU x mg(-1), CAT (93.07 +/- 10.40) U x mg(-1) and the content of GSH (8.91 +/- 1.06) micromol x mg(-1) of cardiomyocytes were also raised by 600 mg x L(-1) l ASS (P < 0.05). CONCLUSION: Taken together, the study implicate that ASS protects cardiomyocytes against oxidative-stress injury induced by H2O2 through reduction of lipid peroxidation and enhancement of the activity of antioxidant defense.