A combination of four effective components derived from Sheng-mai san attenuates hydrogen peroxide-induced injury in PC12 cells through inhibiting Akt and MAPK signaling pathways  

作　　者：CAO Guo-Sheng LI Shao-Xia WANG Yan XU Ying-Qiong LV Yan-Ni KOU Jun-Ping YU Bo-Yang 

机构地区：[1]Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Complex Prescription of TCM, China Pharmaceutical University

出　　处：《Chinese Journal of Natural Medicines》2016年第7期508-517,共10页中国天然药物（英文版）

基　　金：supported by National Natural Science Foundation of China(No.81274004);2011 Program for Excellent Scientific and Technological Innovation Team of Jiangsu Higher Education;the Priority Academic Program Development of Jiangsu Higher Education Institutions

摘　　要：The present study was designed to investigate whether a combination of four effective components derived from Sheng-mai san(SMXZF; ginsenoside Rb1: ginsenoside Rg1: DT^(–1)3: Schizandrol A as 6 : 9 : 4 : 5) could attenuate hydrogen peroxide(H_2O_2)-induced injury in PC12 cells, focusing on the Akt and MAPK pathways. The PC12 cells were exposed to H_2O_2(400 mmol·L^(–1)) for 1 h in the presence or absence of SMXZF pre-treatment for 24 h. Cell viability was measured by MTT assay. The efflux of lactate dehydrogenase(LDH), the intracellular content of malondialdehyde(MDA), the activities of superoxide dismutase(SOD), and caspase-3 were also determined. Cell apoptosis was measured by Hoechst 33342 staining and Annexin V-FITC/PI staining method. The expression of Bcl-2, Bax, cleaved caspase-3, Akt, and MAPKs were detected by Western blotting analyses. SMXZF pretreatment significantly increased the cell viability and SOD activity and improved the cell morphological changes, while reduced the levels of LDH and MDA at the concentrations of 0.1, 1 and 10 μg·m L^(–1). SMXZF also inhibited H_2O_2-induced apoptosis in PC12 cells. Moreover, SMXZF reduced the activity of caspase-3, up-regulated the protein ratio of Bcl-2 and Bax and inhibited the expression of cleaved caspase-3, p-Akt, p-p38, p-JNK and p-ERK1/2 in H_2O_2-induced PC12 cells. Co-incubation of Akt inhibitor or p38 inhibitor partly attenuated the protection of SMXZF against H_2O_2-injured PC12 cells. In conclusion, our findings suggested that SMXZF attenuated H_2O_2-induced injury in PC12 cells by inhibiting Akt and MAPKs signaling pathways, which might shed insights on its neuroprotective mechanism.The present study was designed to investigate whether a combination of four effective components derived from Sheng-mai san （SMXZF; ginsenoside Rbl： ginsenoside Rgl： DT-13： Schizandrol A as 6 ： 9 ： 4 ： 5） could attenuate hydrogen peroxide （HzO2）-induced injury in PC12 cells, focusing on the Akt and MAPK pathways. The PC12 cells were exposed to H202 （400 gmol-L-1） for I h in the presence or absence of SMXZF pre-treatment for 24 h. Cell viability was measured by MTT assay. The effiux of lactate dehydrogenase （LDH）, the intracellular content of malondialdehyde （MDA）, the activities of superoxide dismutase （SOD）, and caspase-3 were also determined. Cell apoptosis was measured by Hoechst 33342 staining and Annexin V-FITC/PI staining method. The expression of Bcl-2, Bax, cleaved caspase-3, Akt, and MAPKs were detected by Western blotting analyses. SMXZF pretreatment significantly increased the cell viability and SOD activity and improved the cell morphological changes, while reduced the levels of LDH and MDA at the concentrations of 0.1, 1 and 10 gg-mL-1. SMXZF also inhibited HzOz-induced apoptosis in PCI2 cells. Moreover, SMXZF reduced the activity of caspase-3, up-regulated the protein ratio of Bcl-2 and Bax and inhibited the expression of cleaved caspase-3, p-Akt, p-p38, p-JNK and p-ERK1/2 in H202-induced PC12 cells. Co-incubation of Akt inhibitor or p38 inhibitor partly attenuated the protection of SMXZF against H202-injured PC12 cells. In conclusion, our findings suggested that SMXZF attenuated H202-induced injury in PC12 cells by inhibiting Akt and MAPKs signaling pathways, which might shed insights on its neuroprotective mechanism.

关 键 词：COMBINATION of EFFECTIVE TCM components Sheng-mai SAN SMXZF PC12 cells Hydrogen peroxide AKT MAPKs 

分 类 号：R965[医药卫生—药理学]