Department of Nephrology, First Affiliated Hospital of China Medical University, Shenyang, China.
Acta Pharmacol Sin. 2011 Aug;32(8):1025-30. doi: 10.1038/aps.2011.74. Epub 2011 Jul 18.
To investigate whether aristolochic acid (AA) induced the apoptosis of human umbilical vein endothelial cells (HUVECs) in vitro and the underlying mechanism.
HUVECs were treated with AA (5, 10 or 20 μg/mL) for 12, 24 and 48 h. Cell viabilities were determined with MTT assay. Hoechst 33258 staining and flow cytometry were used to examine the apoptosis of HUVECs. Western blotting was used to evaluate Akt phosphorylation. Bcl-2 and Bax levels were measured using Western blotting and RT-PCR assays.
Treatment of HUVECs with AA significantly decreased the cell viabilities in dose- and time-dependent manners. Morphological changes of apoptosis were observed in AA-treated cells. AA inhibited Akt activation, which was attenuated by pretreatment of the cells with LY294002 (20 μmol/L) or wortmannin (50 nmol/L). Furthermore, AA reduced Bcl-2 levels and increased Bax levels.
AA induces apoptosis of HUVECs in vitro via the PI3K/Akt signaling pathway and by modulating the ratio Bcl-2 and Bax.
探讨马兜铃酸（AA）是否能诱导人脐静脉内皮细胞（HUVEC）体外凋亡及其机制。
用不同浓度（5、10 或 20μg/ml）AA 处理 HUVEC 12、24 和 48 小时，MTT 法检测细胞活力。Hoechst 33258 染色和流式细胞术检测 HUVEC 凋亡。Western blot 检测 Akt 磷酸化水平。Western blot 和 RT-PCR 检测 Bcl-2 和 Bax 水平。
AA 以剂量和时间依赖方式显著降低细胞活力。AA 处理组细胞出现凋亡形态学改变。LY294002（20μmol/L）或wortmannin（50nmol/L）预处理能减弱 AA 对 Akt 的抑制作用。此外，AA 降低 Bcl-2 水平，增加 Bax 水平。
AA 通过 PI3K/Akt 信号通路和调节 Bcl-2、Bax 比值诱导 HUVEC 体外凋亡。
