CAJ | 학술논문

目的：观察青藤碱( sinomenine,SN)对氧糖剥夺( oxy-gen glucose deprivation,OGD)诱导海马神经元缺血损伤的保护作用并探讨其机制。方法原代培养海马神经元OGD 2 h后复灌24 h(OGD-R),MTT法和LDH试剂盒分别检测各组细胞存活率和乳酸脱氢酶( lactate dehydrogenase,LDH)释放量；Hoechst染色检测细胞凋亡；Westren blot 检测凋亡相关蛋白表达；荧光钙离子成像系统检测[ Ca2+] i ,全细胞膜片钳技术记录酸敏感离子通道( acid-sensing ion channels, ASICs)电流。结果不同浓度SN均能抑制OGD-R诱导的海马神经元存活率降低和LDH释放增多； SN能明显抑制OGD-R诱导的神经元凋亡、Bcl-2/Bax比值的下降、caspase-3表达的增加以及胞内钙浓度升高, SN还能抑制海马神经元ASIC1a电流,也能抑制OGD诱导的ASICs电流增加。结论SN对 OGD诱导海马神经元损伤有保护作用,其机制可能与抑制ASICs通道减轻钙超载相关。
목적：관찰청등감( sinomenine,SN)대양당박탈( oxy-gen glucose deprivation,OGD)유도해마신경원결혈손상적보호작용병탐토기궤제。방법원대배양해마신경원OGD 2 h후복관24 h(OGD-R),MTT법화LDH시제합분별검측각조세포존활솔화유산탈경매( lactate dehydrogenase,LDH)석방량；Hoechst염색검측세포조망；Westren blot 검측조망상관단백표체；형광개리자성상계통검측[ Ca2+] i ,전세포막편겸기술기록산민감리자통도( acid-sensing ion channels, ASICs)전류。결과불동농도SN균능억제OGD-R유도적해마신경원존활솔강저화LDH석방증다； SN능명현억제OGD-R유도적신경원조망、Bcl-2/Bax비치적하강、caspase-3표체적증가이급포내개농도승고, SN환능억제해마신경원ASIC1a전류,야능억제OGD유도적ASICs전류증가。결론SN대 OGD유도해마신경원손상유보호작용,기궤제가능여억제ASICs통도감경개초재상관。
Aim To observe the neuroprotective effect of sinomenine on hippocampal neurons from injury in-duced by oxygen glucose deprivation ( OGD ) and its underlying mechanism. Methods Hippocampal neu-rons were exposed to OGD for 4 h followed by 24 h re-oxygenation ( OGD-R) . Then cell viability was detec-ted by MTT. LDH release was detected by LDH kit. Cell apoptosis was detected by Hoechst stain. The ex-pression of Bax, Bcl-2 and caspase-3 were detected by Western blot. [ Ca2+] i of hippocampal neurons was detected by calcium imaging. Acid-sensing ion chan-nels ( ASICs ) current was detected by patch clamp technique. Results SN increased cell viability and reduced LDH release. SN also inhibited neuron apop-tosis and increased ratio of Bcl-2/Bax and reduced the expression of caspase-3 . OGD-induced increase of [ Ca2+] i was inhibited by SN. Furthermore, SN inhib-ited ASIC1 a current and also inhibited OGD induced increase of ASICs current in hippocampal neurons. Conclusion SN protects hippocampal neurons against OGD-R-induced injury. The inhibitory effect of SN on ASIC1 a and calcium overload was involved in the pro-tective effect of SN.