半枝莲含药血清对肝癌H22细胞凋亡及线粒体膜电位影响.docVIP

半枝莲含药血清对肝癌H22细胞凋亡及线粒体膜电位影响 　 作者：代志军, 王西京, 薛茜, 纪宗正, 刘小旭, 康华峰, 管海涛, 马小斌, 任宏涛 【摘要】 　　目的：探讨中药半枝莲提取物（Scutellaria Barbata extract, ESB）含药血清对肝癌H22细胞凋亡率及线粒体膜电位的影响。方法：体外培养小鼠H22肝癌细胞，以ESB高、中、低剂量含药血清培养液作用于H22细胞，并设立空白血清及5氟尿嘧啶（fluorouracil，5Fu）组（阳性对照组）。应用四甲基偶氮唑盐法检测药物对细胞增殖的抑制作用；透射电镜下观察肿瘤细胞的超微结构变化；碘化丙啶标记，流式细胞术检测肿瘤细胞周期及细胞凋亡情况；采用荧光探针罗丹明123负载，激光扫描共聚焦显微镜下观察H22细胞的荧光强度，分析H22细胞线粒体膜电位的大小。结果：ESB含药血清有一定的抑制H22细胞增殖的作用，高、中剂量组96 h抑制率分别为（47.5±6.4）%和（36.3±4.5）%。透射电镜下可见部分细胞呈典型凋亡形态学改变。流式细胞仪检测结果显示，ESB中、高剂量组有明显的诱导凋亡作用，其凋亡率分别为（3.15±1.12）%和（7.83±2.25）%，与空白组相比差异有统计学意义（Plt;0.01）。空白对照组、5Fu组和ESB低、中、高剂量，H22细胞线粒体膜电位分别为（245.45±67.37）、（127.42±41.35）和（213.68±65.52）、（186.34±56.37）、（142.65±39.44），各组线粒体膜电位与细胞凋亡率呈负相关（r=－0.826, Plt;0.01）。结论：ESB含药血清可有效诱导肿瘤细胞凋亡，其机制可能与降低肿瘤细胞线粒体膜电位有关。 【关键词】 半枝莲 肝癌 细胞凋亡 线粒体膜电位 药物血清 体外研究 　　Objective: To investigate the effects of serum containing Scutellaria Barbata extract (ESB) on apoptosis rate and mitochondrial transmembrane potential (MTP) of liver cancer cell line H22 from mice in vitro. 　　Methods: H22 cells were cultured in vitro and divided into 5 groups: blank control group, lowdose ESB group, mediumdose ESB group, highdose ESB group and fluorouracil (5Fu) group. Methyl thiazolyl tetrazolium assay was utilized to determine the proliferation rates of H22 cells. Cellular morphology was observed under a transmission electron microscope (EM). The rhodamine 123 was used as a fluorescence probe to label the H22 cells, and the fluorescence intensities were observed with a laser scanning confocal microscope. The fluorescence intensity of H22 cells indicated the MTP of H22 cells. 　　Results: The inhibition of serum containing ESB on the proliferation of H22 cells in vitro was observed in a timedependent manner. The typical morphological changes of apoptosis were observed after incubation with ESBcontaining serum in high dose for 48h. The apoptosis rates of blank control group, 5Fu group, lowdose ESB group, mediumdose ESB group and highdose ESB group were (0.51±0.32)%, (11.26±2