蝙蝠葛苏林碱及异构体对PC12细胞缺血性损伤的保护作用

EFFECTS OF DAURISOLINE AND ITS THREE OPTICALISOMERS ON ISCHEMIC INJURY IN CULTURED PHEOCHROMOCYTOMA (PC12) CELLS

用细胞培养术，以ＮａＣＮ加缺糖引起ＰＣ１２细胞损伤作为细胞缺血性损伤的模型，观察蝙蝠葛苏林碱（ＤＳ）及其３个光学异构体对ＰＣ１２细胞缺血性损伤的保护作用；以Ｆｕｒａ２／ＡＭ为Ｃａ２＋的荧光探针，用ＡＲＣＭＭＩＣ阳离子测定系统观察ＤＳ及异构体对ＮａＣＮ引起的Ｃａ２＋内流的抑制作用。结果表明ＤＳ及３个异构体在１０－８～１０－５ｍｏｌ·Ｌ－１的浓度范围内对ＰＣ１２细胞损伤有较明显的保护作用，其机制之一与抑制ＮａＣＮ加缺糖诱发的细胞内游离Ｃａ２＋异常升高有关。

In this study, the protective effects of (-)R·Rdaurisoline and its three optical isomers on ischemic injury in cultured PC12 cells induced by treating cells with NaCN in glucosefree medium were investigated. Cell viability was measured using MTT assay. The results indicated that these compounds, especially (-)S·R and (+)R·S isomers were found substantially to attenuate ischemic injury in PC12 cells in a dosedependent manner. The IC50 values of (-)R·R, (-)S·R, (+)R·S and (+)S·S isomers were shown to be 186×10-6, 24×10-6, 59×10-6 and 90×10-6 mol·L-1, respectively. Intracellular free Ca2+ concentration in PC12 cells was measured using ARCMMIC cation measurement system with Fura2/AM as Ca2+ fluorecent indicator. (-)R·Rdaurisoline and its three optical isomers: (-)S·R, (+)R·S and (-)S·S were found to markedly inhibit the increase of cytosolic free Ca2+ concentration induced by NaCN (20 mmol·L-1) in a dosedependent manner. Their IC50 were found to be 355×10-6, 059×10-6, 129×10-6 and 243×10-6 mol·L-1 respectively. It is suggested that the cytoprotective effects of daurisoline and its isomers were mediated by blocking Ca2+ influx into cells.