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目的探讨二丁酰环磷酰苷(db-cAMP)和氨茶碱分别作用以及联合用药对缺氧葡萄糖剥夺(oxygen-glu-cose deprivation,OGD)条件下大鼠肾上腺嗜铬细胞瘤(PC12)细胞活性和凋亡的影响。方法采用OGD方法建立体外培养的PC12细胞缺氧缺血模型。将PC12细胞分为正常对照组和OGD组,OGD组又分为未用药组,氨茶碱组,db-cAMP组和联合用药组(氨茶碱和db-cAMP)。用四甲基偶氮唑盐(MTT)比色法观察不同药物和不同加药时间(0h,1h,2h,4h,8h,16h)对OGD后PC12细胞活性影响;用Hoechst33342荧光染料测定细胞凋亡。结果经OGD 4h后,OGD组与正常组相比细胞活性降低和凋亡率增加(P<0.05),db-cAMP 1×10-4μmol/L组与未用药组相比细胞活性增高,凋亡率减低(P<0.05);氨茶碱5×10-2μg/L组与未用药组相比细胞活性和凋亡率无明显差别(P>0.05);3×10-2μg/L氨茶碱和1×10-5μmol/L的db-cAMP联合用药组与未用药组相比细胞活性增高,凋亡率减低(P<0.05);联合用药组与1×10-4μmol/L的db-cAMP组相比细胞活性和凋亡率无明显差异(P>0.05);OGD损伤后4h内加药组较未用药组细胞活性明显增加(P<0.05),8h以后加药组与1h和2h加药组相比活性明显降低(P<0.05)。结论提示db-cAMP对缺氧缺血损伤有保护作用,氨茶碱和db-cAMP两药联合使用有协同作用,OGD损伤后4h内给药可提高细胞活性,其中1-2h内给药效果最好。
Objective To investigate the effects of db-cAMP and aminophylline on cell viability of adrenal pheochromocytoma (PC12) cells under oxygen-glucose-deprivation (OGD) And the impact of apoptosis. Methods The hypoxic-ischemic model of PC12 cells cultured in vitro was established by OGD method. PC12 cells were divided into normal control group and OGD group. OGD group was divided into untreated group, aminophylline group, db-cAMP group and combination group (aminophylline and db-cAMP). The effects of different drugs and different dosing time (0h, 1h, 2h, 4h, 8h, 16h) on the activity of PC12 cells after OGD were observed by MTT colorimetric assay. The apoptosis of PC12 cells was detected by Hoechst33342 fluorescent dye Death. Results After OGD for 4 hours, the cell viability decreased and the apoptosis rate increased in OGD group compared with normal group (P <0.05). Compared with non-treated group, db-cAMP 1 × 10-4μmol / L group had higher cell viability and apoptosis (P <0.05). There was no significant difference in cell viability and apoptosis rate between aminophylline 5 × 10-2μg / L group and untreated group (P> 0.05) And 1 × 10-5μmol / L db-cAMP combined with drug group compared with non-treated cells increased activity, decreased apoptosis (P <0.05); combined with 1 × 10-4μmol / L of db-cAMP (P> 0.05). Compared with the untreated group, the cell viability of the treated group increased significantly (P <0.05) within 4 h after OGD injury, Compared with the drug group, the activity was significantly decreased (P <0.05). Conclusions db-cAMP has a protective effect on hypoxic-ischemic injury, aminophylline and db-cAMP synergistic combination of two drugs, OGD damage within 4h after administration can improve cell activity, which 1-2h within the drug delivery the best.