Protective effects of ginsenoside Rb3 on oxygen and glucose deprivation-induced ischemic injury in PC12 cells

Abstract

Aim: To investigate the protective effects of ginsenoside Rb 3 , a triterpenoid saponin isolated from the leaves of Panax notoginseng, on ischemic and reperfusion injury model of PC12 cells and elucidate the related mechanisms. Methods: PC12 cells exposed to oxygen and glucose deprivation (OGD) and restoration (OGD-Rep) were used as an in vitro model of ischemia and reperfusion. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) leakage were used to evaluate the protective effects of ginsenoside Rb 3 . Cellular apoptosis and mitochondrial membrane potential (MMP) were analyzed using flow cytometry. Intracellular calcium ion concentration ([Ca 2+ ] i ) was detected using fluorophotometer system. Caspase-3, -8, and -9 activities were measured using assay kits with an ELISA reader. Western blotting assay was used to evaluate the release of cytochrome c and expression of caspase-3, Bcl-2 and Bax proteins. Results: It was shown that ginsenoside Rb 3 (0.1-10 μmol/L) significantly increased cell viability and inhibited LDH release in a dosedependent manner on the ischemic model. In addition, ginsenoside Rb 3 also significantly inhibited ischemic injury-induced apoptosis, [Ca 2+ ] i elevation, and decrease of MMP. Meanwhile, pretreatment with ginsenoside Rb 3 significantly induced an increase of Bcl-2 protein expression and a decrease of cytosolic cytochrome c, cleaved-caspase 3 and Bax protein expression, the caspase-3, -8, and -9 activity were also inhibited. Conclusion:The results indicated that ginsenoside Rb 3 could markedly protected OGD-Rep induced ischemic injury and the mechanisms maybe related to its suppression of the intracellular Ca 2+ elevation and inhibition of apoptosis and caspase activity. Ginsenoside Rb 3 could be a promising candidate in the development of a novel class of anti-ischemic agent.

Pre-treatment of a single high-dose of atorvastatin provided cardioprotection in different ischaemia/reperfusion models via activating mitochondrial KATP channel

“…The activity of LDH in culture medium could also be used as a marker of cell injury. LDH is a stable cytoplasmic enzyme that is present in most cells, and is rapidly released into the culture medium following plasma membrane damage (Zhu et al, 2010). As shown in Fig.…”

β-Asarone protects PC12 cells against OGD/R-induced injury via attenuating Beclin-1-dependent autophagy

“…It can acquire neuron-like properties when exposed to nerve growth factor (NGF) [12] . Therefore, it is widely used as an in vitro model of cerebral ischemia/reperfusion [13,14] . Intracellular free calcium concentration ([Ca 2+ ] i ) and mitochondrial membrane potential (MMP) are important indicators to reflect cell injury.…”

Involvement of N-Methyl-D-Aspartic Acid Receptor and DL-α-Amino-3-Hydroxy-5- Methyl-4-Isoxazole Propionic Acid Receptor in Ginsenosides Rb1 and Rb3 against Oxygen-Glucose Deprivation-Induced Injury in Hippocampal Slices from Rat

“…Ginsenoside Rb1 could reduce the cortical infarct volume by approximately 50% and decrease the number of TUNEL-positive cells in the hippocampal CA1 region after ischemia [16]. Zhu et al [17] reported that ginsenoside Rb3 could markedly protect OGD-induced ischemic injury on PC12 cells. In our previous studies, we found that ginsenoside Rb3 exhibited the better neuroprotective effect than ginsenoside Rb1 in rat hippocampal slices using an in vitro ischemic model [6], but the underlying mechanism of action was still unclear.…”