Zhang Yanmin, He Langchong, Zhou Yali
School of Medicine, Xi'an Jiaotong University, No. 76, Yanta Weststreet, #120, Xi'an, Shaanxi Province 710061, PR China.
Phytomedicine. 2008 Jan;15(1-2):112-9. doi: 10.1016/j.phymed.2007.09.021. Epub 2007 Oct 31.
The present study was to evaluate the effects of taspine isolated from Radix et Rhizoma Leonticsi on the growth and apoptosis of human umbilical vein endothelial cell (HUVEC) line by MTT and flow cytometer, respectively. At the same time, a series of changes were observed in HUVEC treated by taspine, including microstructure, protein expression of bax, bcl-2 and VEGF. The change of microstructure was observed by transmission electron microscope (TEM). The protein expression of bax and bcl-2 was detected by immunohistochemistry (IHC), and VEGF protein secreted was determined by enzyme-linked immunosorbent assay (ELISA). The results showed taspine could inhibit growth and induce apoptosis of HUVEC in a dose-dependent manner. Cell cycle was significantly stopped at the S phase. Under electronic microscope, the morphology of HUVEC treated with taspine showed nuclear karyopycnosis, chromatin agglutination and typical apoptotic body. Bcl-2 and VEGF expressions were decreased and bax expression was increased. All these results demonstrate that taspine has an inhibitory effect on growth of HUVEC and can induce its apoptosis.
本研究旨在分别通过MTT法和流式细胞仪评估从地锦草中分离得到的塔斯品碱对人脐静脉内皮细胞（HUVEC）系生长和凋亡的影响。同时，观察塔斯品碱处理HUVEC后的一系列变化，包括微观结构、bax、bcl-2和VEGF的蛋白表达。通过透射电子显微镜（TEM）观察微观结构变化。采用免疫组织化学（IHC）检测bax和bcl-2的蛋白表达，并用酶联免疫吸附测定（ELISA）法测定分泌的VEGF蛋白。结果表明，塔斯品碱可抑制HUVEC的生长并诱导其凋亡，呈剂量依赖性。细胞周期在S期显著停滞。在电子显微镜下，经塔斯品碱处理的HUVEC形态表现为核固缩、染色质凝集和典型的凋亡小体。Bcl-2和VEGF表达降低，bax表达增加。所有这些结果表明，塔斯品碱对HUVEC的生长具有抑制作用，并可诱导其凋亡。
