原儿茶酸对体外培养的神经干／祖细胞增殖及凋亡的影响

Studies on the effects of protocatechuic acid on cell proliferation and apoptosis in cultured neural stem/progenitor cells

目的观察原儿茶酸对体外培养的神经干/祖细胞的增殖和自发性凋亡的影响。方法神经干/祖细胞取自胚胎14 d小鼠海马组织,以悬浮方式在培养器皿中培养成"神经球"。CCK-8试剂盒检测细胞的生存力,B rdU染色标记分析细胞的增殖。结果培养4 d后,神经干/祖细胞发生了自发性凋亡,原儿茶酸(0.06 mmo.lL-1)有效地减少了该种凋亡,将细胞的存活率提高至对照组的140.3%±9.2%;细胞培养4 d和7 d,原儿茶酸(0.06 mmo.lL-1)使细胞内ROS水平分别下降至对照组的43.8%±4.7%和54.5%±6.1%,caspase-3活性下降至对照组的70.6%±4.4%和62.3%±5.5%。结论原儿茶酸剂量和时间依赖地提高了神经干/祖细胞的生存力并且刺激了细胞的增殖。

Aim To investigate the effects of PCA on cultured neural stem/progenitor cells （NS/PCs） proliferation and basal apoptosis. Methods NS/PCs obtained from 14-day-old rat embryos were propagated as neurospheres and cultured under normal condition with or without PCA for the indicated time. The cell viability was determined by Cell Counting Kit-8 （CCK-8） test, while cell proliferation was assayed by bromodeoxyuridine （BrdU） labeling. Results After 4 days, basal apoptotic cells were detected and PCA significantly reduced the levels of apoptosis of NS/PCs. Treatment with 0.06 mmol· L^-1 PCA caused 140.32% ±9. 17% increase in cell viability compared with the control. Furthermore, on day 4 and day 7, the reactive oxygen species （ROS） level in NS/PCs was depressed by 43.8% ±4.7% and 54. 5% ±6. 1% ,respectively, and caspase-3 activity was decreased by 70. 6% ± 4.4% and 62.3% ± 5.5% respectively. Conclusions PCA can increase the cellular viability of NS/PCs and stimulate cell proliferation in a doseand time-dependent manner.