半枝莲含药血清对肝癌H22细胞凋亡及线粒体膜电位的影响.doc

半枝莲含药血清对肝癌 H22 细胞凋亡及线粒
体膜电位的影响

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， 凋亡率分别为 3.15±1.12） 和 7.83
其 （ %（

±2.25）%，与空白组相比差异有统计学意义（P<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
、 ，

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关（r=－0.826, P<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.

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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±

2.97)%, (1.07 ± 0.46)%, (3.15 ± 1.12)%, (7.83 ± 2.25)%

respectively. ESB could reduce the MTP of H22 cells from mice

as compared with the untreated group. The MTPs of the blank

control group, 5 Fu group, and low , medium and high

dose ESB groups were (245.45±67.37), (127.42±41.35), (213.68

±65.52), (186.34±56.37) and (142.65±39.44) respectively,

which were negatively correlated with the apoptosis rates.

Conclusion: ESB containing serum effectively induces

apoptosis, which may be related to the decrease of MTP in H22

cells.

Keywords: Scutellaria barbata; liver cancer; apoptosis;

mitochondrial transmembrane potential; drug containing serum;

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