基于析因设计的川芎嗪和阿魏酸配伍抗PC12氧糖剥夺损伤的交互作用研究

Compatibility of Tetramethylpyrazine and Ferulic Acid Interaction of anti PC12 damage based on Factorial Design

目的观察川芎嗪和阿魏酸配伍对PC12糖氧剥夺(OGD)损伤的预保护作用,优化配比组合。方法体外培养PC12,考虑A(是否OGD造模)、B(川芎嗪0.5、1.0、2.0μmol/L)及C(阿魏酸5、10、20μmol/L)3个因素,采用完全随机分组的2×3×3析因设计。倒置相差显微镜观察造模前后细胞形态;MTS测细胞活力;采用乳酸脱氢酶(LDH)法测定细胞损伤,并进行析因分析。结果与对照组比较,模型组细胞变圆或肿胀,突起回缩或消失,细胞膜褶皱或破损,胞体折光性差,细胞数目明显减少,细胞活力降低;与模型组比较,加药组细胞损伤得到不同程度的改善,细胞活力增加,川芎嗪2μmol/L+阿魏酸20μmol/L改善作用最明显。A、B、C 3因素均对LDH值具有显著影响(P<0.01),且B、C间存在交互作用(P<0.01),以川芎嗪2μmol/L、阿魏酸20μmol/L配比时,LDH值最低。结论川芎嗪与阿魏酸合用对PC12剥夺糖氧损伤具有预保护作用,川芎嗪与阿魏酸配比具有交互作用,最佳配比为2∶20。

Objective To investigate the protective effects of monomer tetramethylpyrazine（TMP） and ferulic acid（FA） on PC12 with oxygen-glucose deprivation（OGD） injury and optimize the combination ratio. Method PC12 cells were cultured in vitro, and A（whether OGD）, B（0.5, 1.0, 2.0 μmol/L TPM）, and C（5, 10, 20 μmol/L FA） were considered as three factors. Adopting completely randomized 2×3×3 factorial design, using inverted phase contrast microscope to observe cell morphology, and MTS and lactic dehydrogenase method were used to detect cell activity. Results Compared with the control group, the cells in the model group became round or swollen, the processes were retracted or disappeared, the cell membrane was folded or damaged, the refraction of cell body was poor, the number of cells was significantly reduced, and the cell viability was decreased; Compared with the model group, the cell injury was improved in varying degrees and the cell viability was increased in the treated group, and 2 μmol/L TMP ＋ 20 μmol/L FA had the most obvious positive effect. Factor A, B, and C had a significant effect on LDH（P〈0.01）, and there is an interaction between factor B and C（P〈0.01）, and when the dose of TMP and FA respectively was 2 μmol/L and 20 μmol/L, the value of LDH was lowest. Conclusion TMP combined with FA had a protective effect on PC12 cells with OGD injury. There is an interaction between TMP and FA, and their best ratio is 2：20.