Li Jun, Li Bin, Zhou Wen-jiang, Zhao Fan-gui, Li Da-jin, Wang Wen-jun
Department of Integrated Traditional Chinese and Western Medicine, Fudan University, Shanghai 200011, China.
Zhong Xi Yi Jie He Xue Bao. 2010 May;8(5):472-9. doi: 10.3736/jcim20100512.
To investigate the effects of medicated rat serum containing Gengnianchun (GNC) decoction and its protection to pheochromocytoma cells (PC12 cells) from amyloid beta (Abeta)(25-35)-insulted apoptosis and to find the possible mechanism.
Medicated rat serum was prepared by administering ovariectomized Sprague-Dawley (SD) rats with GNC decoction. The effects of medicated rat serum on viability of PC12 cells were evaluated by cell counting kit-8 (CCK-8) assay. The PC12 cells were cultured with different doses of Abeta(25-35) to induce a model of Alzheimer's disease in vitro. Then, the protective effects of medicated rat serum on Abeta(25-35)-insulted PC12 cells were evaluated by using CCK-8 assay to detect the cell viability, using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry to detect cell apoptosis rate and using Western blotting assay to analyze the expressions of Bcl-2, Bax and active caspase-3 proteins.
PC12 cells cultured with 20% medicated rat serum containing GNC decoction for 24 h or 48 h had higher viability than those cultured with normal culture medium (P<0.05). After 24- or 48-hour treatment of different concentrations of Abeta(25-35), cell viabilities were all decreased as compared with normal medium (P<0.05). Cells underwent apoptosis, which showed the neurotoxicity of Abeta(25-35). The cell apoptosis induced by Abeta 25-35 was significantly decreased in PC12 cells which were pretreated with 20% medicated rat serum or nerve growth factor (NGF) according to CCK-8 assay and Annexin V-FITC/PI flow cytometry (P<0.05). The ratio of Bax expression to Bcl-2 expression and the expression of active caspase-3 were decreased in the cells treated with medicated serum or NGF as compared with the cells cultured with Abeta(25-35) only.
The GNC-medicated rat serum at concentration of 20% can promote viability of Abeta(25-35)-insulted PC12 cells and decrease the cell apoptosis by regulating the expressions of Bcl-2, Bax and active caspase 3.
研究含更年春（GNC）水煎剂含药血清对β淀粉样蛋白（Aβ）（25 - 35）损伤的嗜铬细胞瘤细胞（PC12细胞）凋亡的影响及其保护作用，并探讨其可能机制。
采用去卵巢的Sprague-Dawley（SD）大鼠灌胃GNC水煎剂制备含药血清。通过细胞计数试剂盒-8（CCK-8）法评估含药血清对PC12细胞活力的影响。将PC12细胞用不同剂量的Aβ（25 - 35）培养以体外诱导阿尔茨海默病模型。然后，通过CCK-8法检测细胞活力、Annexin V-异硫氰酸荧光素（FITC）/碘化丙啶（PI）流式细胞术检测细胞凋亡率以及Western印迹法分析Bcl-2、Bax和活化的半胱天冬酶-3蛋白的表达，评估含药血清对Aβ（25 - 35）损伤的PC12细胞的保护作用。
用含20%含GNC水煎剂的含药血清培养PC12细胞24小时或48小时后，其活力高于用正常培养基培养的细胞（P<0.05）。用不同浓度的Aβ（25 - 35）处理24小时或48小时后，与正常培养基相比，细胞活力均降低（P<0.05）。细胞发生凋亡，显示出Aβ（25 - 35）的神经毒性。根据CCK-8法和Annexin V-FITC/PI流式细胞术检测，用20%含药血清或神经生长因子（NGF）预处理的PC12细胞中，Aβ 25 - 35诱导的细胞凋亡明显减少（P<0.05）。与仅用Aβ（25 - 35）培养的细胞相比，用含药血清或NGF处理的细胞中Bax表达与Bcl-2表达的比值以及活化的半胱天冬酶-3的表达均降低。
20%浓度的GNC含药血清可促进Aβ（25 - 35）损伤的PC12细胞的活力，并通过调节Bcl-2、Bax和活化的半胱天冬酶3的表达降低细胞凋亡。
