Protective effect of acanthopanax senticosus saponins on anoxia-damaged motoneurons in rat spinal cord in vitro / 第二军医大学学报

ABSTRACT

Objective: To investigate whether acanthopanax senticosus saponins(ASS) has a protective effect on anoxia-damaged rat spinal cord motoneurons(SMNs) in vitro and to reveal the possible mechanism of this effects. Methods: SMNs were obtained from the spinal cord of embryonic day 15 rats and were cultured in vitro. The cultured cells were immunohistochemically identified and were subjected to anoxia exposure to establish apoptosis model. In this study, SMNs were divided into the following 4 groups normal control group;anoxia-induced apoptosis group;ASS pre-treated group, SMNs were treated with ASS (50 μg/ml) 24 h before anoxia exposure; and glial-cell-line-derived neurotrophic factor(GDNF) pre-treated group, SMNs were treated with GDNF(0.1 μg/ml) 24 h before anoxia exposure. The morphology of SMNs was observed with phase-contrast microscope and electron microscope; the viability of SMNs was detected with MTT method; and the influence of ASS on the stablility of SMNs membrane was observed through detection of lactate dehydrogenase (LDH) level in the extracellular solution. The influence of ASS on the expression of HIF-1α in anoxia-damaged SMNs was studied by Western blot analysis. Results: The morphological damage of SMNs in ASS and GDNF pre -treated groups was slighter than that in anoxia-induced apoptosis group. The results showed that the viability of SMNs was higher in ASS pre-treated group (0.21±0.028) compared with that in anoxia-induced apoptosis group (0.15 ± 0.012) (P < 0.05), while the level of LDH was lower in ASS pre-treated group (28.6±1.309) than that in anoxia-induced apoptosis group (40.7±1.885) (P<0.01). The expression of HIF-1α in ASS pretreated group was the highest (1.15±0.016) (P<0.01) and that in anoxia-induced apoptosis group (0.72±0.027) was higher than that in the control group (0.16±0.003) (P<0.01). Protective effect of ASS on anoxia-damaged SMNs in rats was similar to that of GDNF. Conclusion: AS S can increase the viability of hypoxia-damaged SMNs in vitro and has obvious protective effects on them, which may be related to the enhanced stability of cell membrane and the up-regulation of HIF-1α expression.