Tang Minke, Zhang Juntian
Pharmacology Department 2, Institute of Materia Medica, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing.
Jpn J Pharmacol. 2002 Apr;88(4):422-7. doi: 10.1254/jjp.88.422.
To observe the effect of salvianolic acid-B (SalB) against the cytoxicity of amyloid beta peptide (A-beta)(25-35) to PC12 cells, the cells were incubated with A-beta, and the cytoxicity was investigated by MTT, flow cytometry and a cell free apoptotic system. The expression of prostate apoptotic response-4 (Par-4) was detected by Western blot. Aged A-beta 10 micromol/L significantly inhibited the MTT reduction of PC12 cells, SalB1 micromol/L inhibited the toxicity induced by A-beta. In flow cytometric analysis, PC12 cells treated with A-beta exhibited degraded DNA content characteristic of apoptosis cells (1.53% vs 19.9%). PC12 cells pretreated with SalB (10 nmol/L, 100 nmol/L, 1 micromol/L) manifested relatively low proportion of apoptosis (15.7%, 13.5%, 11.8%, respectively). SalB (10 nmol/L - 1 micromol/L) when added at the beginning of the cell free apoptotic reaction had no apparent effect on the nuclei apoptosis. Pretreatment of PC12 cells with SalB largely prevented the increase in Par-4 expression of the cells when they were exposed to A-beta. The results suggest that Par-4 is involved in the protective effect of SalB against A-beta-induced damage in PC12 cells.
为观察丹酚酸B(SalB)对β淀粉样肽(A-β)(25 - 35)诱导PC12细胞毒性的影响,将细胞与A-β共同孵育,采用MTT法、流式细胞术及无细胞凋亡体系研究细胞毒性。通过蛋白质免疫印迹法检测前列腺凋亡反应蛋白4(Par-4)的表达。10 μmol/L老化的A-β显著抑制PC12细胞的MTT还原,1 μmol/L的SalB可抑制A-β诱导的毒性。在流式细胞术分析中,用A-β处理的PC12细胞呈现出凋亡细胞特征性的DNA降解含量(1.53%对19.9%)。用SalB(10 nmol/L、100 nmol/L、1 μmol/L)预处理的PC12细胞凋亡比例相对较低(分别为15.7%、13.5%、11.8%)。在无细胞凋亡反应开始时加入SalB(10 nmol/L - 1 μmol/L)对细胞核凋亡无明显影响。用SalB预处理PC12细胞可在很大程度上阻止细胞在暴露于A-β时Par-4表达的增加。结果表明,Par-4参与了SalB对PC12细胞中A-β诱导损伤的保护作用。