四味芳香开窍药的挥发性成分对缺血缺氧PC12细胞及细胞内Ca2+的影响

目的研究四味芳香开窍药的挥发性成分对连二亚硫酸钠(Na2S2O4)致PC12细胞缺血缺氧损伤及细胞内Ca2+的影响。方法体外实验采用5mmol/L的Na2S2O4导致PC12细胞缺血缺氧性损伤,然后观测细胞形态;用二甲基噻唑二苯基四唑溴盐(MTT)比色法,考察水蒸汽蒸馏法提取的麝香、苏合香、安息香挥发性成分及冰片对缺血缺氧PC12细胞活性的影响;并按照试剂盒要求测定模型细胞内Ca2+的含量。结果麝香4、8、16μg/mL,冰片8、16μg/mL,苏合香20、40μg/mL及安息香16μg/mL组均能显著升高模型细胞的吸光度(A)值(P<0.05,P<0.01),显著抑制Na2S2O4对PC12细胞的损伤;苏合香40、80μg/mL,安息香4、8、16μg/mL组均能显著降低模型细胞内Ca2+含量(P<0.05,P<0.01);麝香、冰片有降低模型细胞内Ca2+含量的趋势。结论麝香、苏合香、安息香的挥发性成分及冰片均有保护缺血缺氧PC12细胞的作用,其机制可能与降低细胞内Ca2+的含量,防止钙超载有关。

Effects of volatile constituents of four resuscitation-inducing aromatic herbs on oxygen glucose deprivation in PC12 cells and intracellular Ca2+

Objective To investigate the effects of volatile constituents of four resuscitation-inducing aromatic herbs on sodium dithionote(Na2S2O4)-induced oxygen glucose deprivation and Ca2+ in differentiated rat pheochromocytoma(PC12) cells.Methods In vitro research,oxygen glucose deprivation model of PC12 cells was established by exposure to Na2S2O4 of 5 mmol/L.The protective effects of volatile constituents of Moschus,Styrax,Benzoinum extraction by water-vapor distilling and Borneolum Syntheticum on the model were evaluated by cytomorphology and colormetric MTT assay.The concentration of intracellular Ca2+ was assayed by the determination system.Results Moschus(4 μg/mL,8 μg/mL and 16 μg/mL),Borneolun Syntheticum(8 μg/mL and 16 μg/mL),Styrax(20 μg/mL and 40 μg/mL) and Benzoinum(16 μg/mL) markedly increased the optical density at 490 nm tested by colorimetric MTT assay(P<0.05,P<0.01) and showed their significant effects in protecting PC12 cells against Na2S2O4-induced apoptosis.Styrax(40 μg/mL and 80 μg/mL) and Benzoinum(4 μg/mL,8 μg/mL and 16 μg/mL) markedly attenuated the concentration of intracellular Ca2+(P<0.05,P<0.01).Moschus and Borneolun Syntheticum showed the tendency of reducing the concentration of intracellular Ca2+.Conclusion The volatile constituents of Moschus,Styrax,Benzoinum and Borneolum Syntheticum can markedly protect PC12 cells from oxygen glucose deprivation injury through attenuating the concentration of intracellular Ca2+ and preventing calcium overload.