苦参素通过p38/JNK信号途径对海马神经元凋亡的影响  

Effect of oxymatrine on apoptosis of hippocampal neurons by p38/JNK signaling pathway

作　　者：蔡艳[1] 蒋伟[2] 周爱玲[3] 赵敏[3] 徐玲[3] CAI Yan JIANG Wei ZHOU Ai-ling ZHAO Min XU Ling(Nantong Hospital of Traditional Chinese Medicine, Nantong 226001, China Department of Technology and Industry, Nantong University, Nantong 226001, China School of Medicine, Nantong University, Nantong 226001, China)

机构地区：[1]南通市中医院,江苏南通226001 [2]南通大学科技与产业处,江苏南通226001 [3]南通大学医学院,江苏南通226001

出　　处：《中国中药杂志》2017年第4期731-738,共8页China Journal of Chinese Materia Medica

基　　金：南通市应用研究科技计划项目(BK2013007)

摘　　要：探讨苦参素(oxymatrine,OMT)对海马神经元凋亡的影响及其机制。采用MTT法测定OMT对海马神经元存活率的影响;生物化学法测定OMT对LPS诱导的海马神经元乳酸脱氢酶(lactate dehydrogenase,LDH)释放率的影响;Hochest 33342染色观察海马神经元凋亡形态;实时定量PCR(Real-time quantitative PCR,RT-q PCR)方法检测海马神经元中Bax,Bcl-2,Caspase-3 mRNA的表达;蛋白免疫印迹法(Western blot,WB)检测海马神经元中p38,p-p38,JNK,p-JNK,Bax,Bcl-2和Caspase-3的表达。结果发现,不同剂量的OMT(0.37~6.0 g·L^(-1))和海马神经元共培养24 h,海马神经元生长状态良好;LPS刺激后,海马神经元LDH释放率增加(P<0.01),JNK和p38的磷酸化水平明显增加(P<0.01),Bax/Bcl-2的比例及Caspase-3的表达上调(P<0.01),海马神经元凋亡增加。OMT预处理后,能明显降低海马神经元经LPS刺激后的LDH释放(P<0.05或P<0.01),减少p38和JNK磷酸化水平,降低Bax/Bcl-2的比例及Caspase-3的表达(P<0.01),海马神经元凋亡减少。综上,OMT能降低经LPS激活的p38和JNK磷酸化水平,下调Bax/Bcl-2的比例和Caspase-3的表达,从而抑制海马神经元凋亡。OMT通过p38/JNK信号途径抑制海马神经元的凋亡。To investigate the effect and mechanism of oxymatrine（ OMT） on hippocampal neurons apoptosis. Effect of OMT on survival of hippocampal neurons was measured by MTT. Effect of OMT on LPS-induced lactate dehydrogenase（ LDH） release rate in hippocampal neurons was measured by biochemical methods. Hoechst 33342 staining was used to observe the apoptotic morphology of hippocampal neurons. The mRNA expression levels of Bax,Bcl-2,and Caspase-3 were detected by Real-time quantitative PCR（ RTq PCR）,and the protein expression levels of p38,p-p38,JNK,p-JNK,Bax,Bcl-2 and Caspase-3 were detected by Western blot. The results showed that,hippocampal neurons all grew well after treatment by different doses（ 0. 37-6. 0 g·L- 1） of OMT for 24 h. Stimulation from LPS increased the release of LDH（ P〈0. 01）,improved the JNK and p38 phosphorylation levels（ P〈0. 01）,increased the proportion of Bax / Bcl-2 and the expression of Caspase-3（ P〈0. 01）,and promoted the apoptosis of hippocampal neurons. OMT pretreatment could significantly reduce the release of LDH induced by LPS stimulation（ P〈0. 05 or P〈0. 01）,reduce the p38 and JNK phosphorylation,decrease the expression of Caspase-3 and Bax / Bcl-2（ P〈0. 01）,and diminish the apoptosis of hippocampal neurons. In conclusion,OMT could reduce the LPS-induced phosphorylation of p38 and JNK,down-regulate the Bax / Bcl-2 ratio and expression of Caspase-3,thus inhibiting apoptosis of hippocampal neurons. The mechanism may be associated with p38 / JNK signaling pathway.

关 键 词：苦参素 海马神经元 细胞凋亡 p38/JNK 

分 类 号：R285[医药卫生—中药学]