摘要
目的观察大黄酚对缺氧PC12细胞损伤的保护作用;方法培养PC12细胞,建立缺氧致神经细胞损伤模型;缺氧前后四甲基偶氮唑盐法(MTT)检测PC12细胞增殖活性、光镜观察PC12细胞形态、测定上清液中乳酸脱氢酶(LDH)活性、早期癌基因表达产物(c-fos)荧光免疫组化表达和逆转录酶聚合酶链反应(RT-PCR)分析神经型一氧化氮合酶(nNOS)、诱导型一氧化氮合酶(iNOS)mRNA的表达;结果大黄酚明显改善缺氧PC12细胞的活力,对损伤细胞形态有改善作用,使损伤细胞的培养上清液中的LDH明显减少;c-fos表达减少,PCR结果分析显示nNOS mRNA在缺氧早期表达。iNOSmRNA主要在缺氧晚期表达。结论本缺氧模型能够引起PC12细胞凋亡现象,大黄酚能减轻PC12细胞缺氧损伤,对神经元细胞有保护作用。
Objective To observe the effect of chrysophanol on hypoxic injured nerve cells. Methods The hypoxia-induced nerve cells injury model was established. PC12 cell proliferation was observed before and after hypoxia by multiply-table tournament （M＇IT） assay and morphology of PC12 cells were observed by light microscopy. The supematant measured superoxide dismutase （LDH） activity was detected. Expressions of neuronal nitric oxide synthase （nNOS） mRNA and inducible nitric oxide synthase （iNOS） mRNA were analyzed by fluorescent inxnunohistochemistry and reverse transcription polymerase chain reaction （RT-PCR） analysis. Results Chrysophanol increased obviously the hypoxic injured PC12 cells viability, and improved the morphology of injured cells; lactic dehydrogenase （LDH） in the supernatant was significantly decreased and c-fos expression was decreased. PCR analysis showed that nNOS mRNA was expressed in early stage of hypoxia, while iNOS mRNA in late stage of hypoxia. Conclusion ~ris hypoxia model can induce the apoptosis of PC12 cells and chrysophanol can reduce the hypoxic injury, which plays a protective role on neurons.
出处
《中华神经外科疾病研究杂志》
CAS
2013年第6期517-522,共6页
Chinese Journal of Neurosurgical Disease Research
基金
甘肃省自然科学基金资助项目(096RJZA095)
