二丁酰环磷酰苷和氨茶碱对PC12细胞缺氧葡萄糖剥夺保护作用初探

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—glueose 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OGD4h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。

The protective effect of db- cAMP and aminophylline to OGD injured PC12 cell.

Objectives： To investigate the influences of db -cAMP, aminophylline and db- cAMP associated with aminophylline on the cells activities and cells apoptosis effects of PC12 cells. Methods ： PC12 cells were used to establish hypoxic - ischemic model by oxygen -glucose deprivation （OGD） method. PC12 cells were divided into OGD group and normal control group. OGD group was then divided into control group, aminophylline group, db -cAMP group and aminophylline associated with db -cAMP group. Cells activities were observed by MTT colorimetry. Hoechst33342 was used to detect cells apoptosis. Results： 4 hours after OGD, cells activities of OGD group were decreased significantly （ P = 0. 000）, while apoptosis increased dramatically （ P = 0. 000） as compared to the control group. Cells activities of db - cAMP （ 1 × 10^-4 μmoL/L） group increased significantly （ P 〈 0. 05） and apoptosis decreased dramatically （ P 〈 0. 05） as compared to the control group. There were no significant difference between aminophylline （5 × 10^-2 μg/L） group and control group in the influence of cells activities and apoptosis. Cells activities of aminophylline （3 × 10^-2 μg/L） associated with db -cAMP （1 ×10^-5μmol/L） group increased significantly （ P 〈0.05） and apoptosis decreased dramatically （ P 〈0. 05） as compared to the control group. There was no significant difference between db- cAMP （1 × 10^-4μmol/L） group and aminophylline （3 × 10^ -2 μg/L） associated with db - cAMP （ 1 × 10^-5 μmol/L） group in the influence of cells activities and apoptosis （ P 〉 0. 05）. Compared to the control group, the cells activities increased significantly as drugs were administrated within 4 hours after OGD injury （ P 〈0. 05）, the cells activities of the lh and 2h groups was significantly higher than the group of 8h （ P 〈0.05） Conclusion： db -cAMP could improve the cells activities of PC12 cells after ODG. The dose of db -cAMP can be reduced when combined with aminophylline, db - cAMP has benefit effectives to the hypoxic - ischemic PC12 ceils. There was a synergistic effect when aminophylline and db - cAMP were used together. It was the best that extractive astragalus cromarile was used 1 - 2h after OGD injury of PC12 cells. Drugs protected PC12 cells when they were administrated within 4 hours after OGD injury.