| 中药片仔癀胶囊对骨肉瘤U-2OS细胞诱导
凋亡的作用 |
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Received:December 09, 2008
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| 期刊信息：《中国骨伤》2009年22卷，第4期，第265-268页 |
| DOI：doi:10.3969/j.issn.1003-0034.yyyy.nn.zzz |
| 基金项目:卫生部科学研究基金(WKJ2005-2-014;WKJ2008-2-42);福建省高等学校新世纪优秀人才支持计划闽教科[(2006)35号] |
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目的：研究中药片仔癀对骨肉瘤U-2OS细胞的诱导凋亡作用。
方法：①采用1月龄SD大鼠16只，雌雄各半，体重120 g左右，随机分为2组(空白血清组和片仔癀治疗组),每组8只。空白血清组大鼠灌服PBS,片仔癀治疗组大鼠灌服中药片仔癀溶液。②人骨肉瘤U-2OS细胞培养于含10%胎牛血清，青霉素100 IU/ml,链霉素100 μg/ml的RPMI 1640培养液中，置于37 ℃、5%CO2饱和湿度的细胞培养箱中培养。用MTT法检测不同含药血清浓度对骨肉瘤U-2OS细胞增殖的影响，筛选对骨肉瘤U-2OS细胞作用最佳的片仔癀含药血清浓度。③在倒置相差显微镜和电镜下观察骨肉瘤U-2OS细胞形态。④ TUNEL原位末端标记法检测细胞的凋亡。⑤ 提取基因DNA,琼脂糖凝胶电泳，以DNA琼脂糖凝胶电泳和透射电镜检测细胞凋亡。
结果：① 20%片仔癀含药血清对骨肉瘤细胞的增殖抑制作用最佳。②倒置相差显微镜观察可见空白血清组细胞呈梭形贴壁生长，在20%片仔癀胶囊含药血清组，可见部分细胞体积变小、变圆，细胞膜完整但出现发泡现象，折光性增强。随作用时间延长，变圆细胞逐渐增多，部分贴壁细胞皱缩、变圆、脱落，可见坏死细胞及碎片。③ 20%片仔癀胶囊含药血清组TUNEL检测阳性。④ DNA Ladder电泳结果：含药血清组细胞基因DNA凝胶电泳出现明显的梯状带。⑤ 透射电镜观察染色质沿皱缩的核膜下凝聚，细胞表面出现凋亡小体。
结论：中药片仔癀胶囊对骨肉瘤U-2OS细胞有抑制增殖和诱导凋亡作用。 |
| [关键词]:细胞凋亡 中药 片仔癀 骨肉瘤U-2OS细胞 |
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| Apoptosis induction of traditional Chinese herb Pianzihuang (片仔癀) in human osteosarcoma U-2OS cells |
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Abstract:
Objective: To study the effect of apoptosis induction of Pianzihuang in human osteosarcoma U-2OS cells.
Methods: ①Sixth SD rats were randomly divided into group A (8 rats) and group B (8 rats) in the study. Among them,8 rats (group A) were treated with PBS and 8 rats(group B) treated with Pianzihuang. ②Human osteosarcoma U-2OS cells were cultured in nutrient solution (including fetal calf serum of 10%,penicillin of 100 IU/ml,streptomycin of 100 μg/ml),the environic condition in saturated humidity 5%CO2,37 ℃。 The osteosarcoma U-2OS cell proliferation effect by different saturation of blood serum was measured by MTT method. And the optimal concentration of the serum(including Pianzihuang) could be obtained. ③The morphous of osteosarcoma U-2OS cell were observed by inverted phase contrast microscope and electron microscope. ④Apoptosis cell was detected by TUNEL in situ end labeling. ⑤Extracted DNA,agarose gel electrophoresis(AGE) to detect apoptosis cell by transmission electron microscope (TEM).
Results: ①It was best that osteosarcoma U-2OS cell proliferation effect by 20% of the concentration of the serum (including Pianzihuang). ②On inverted phase contrast microscope showed that the cells of group A were fusiform adherence growth;the cyton transform shrind and circle in 20% of the concentration of the serum(including Pianzihuang). Follow lasting time,the cell turning circle gradually increased. Necrosis cell and fragment were found by microscope. ③The detected result of apoptosis cell in 20% of the concentration of the serum(including Pianzihuang) was positive by TUNEL assays. ④AGE showed “ladder” strand of DNA,a special phenomenon of cellular apoptosis. ⑤Chromatin aggregated along corrugativus caryotheca and apoptotic body appeared on cell surface by observation of TEM.
Conclusion: Pianzihuang can inhibit proliferation and induce apoptosis for osteosarcoma U-2OS cell |
| KEYWORDS:Apoptosis Chinese herbal drugs Pianzihuang (片仔癀) Osteosarcoma U-2OS cells |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 张俐,于波,林建华.中药片仔癀胶囊对骨肉瘤U-2OS细胞诱导
凋亡的作用[J].中国骨伤,2009,22(4):265~268 |
| 英文格式: | ZHANG Li,YU Bo,LIN Jian-hua.Apoptosis induction of traditional Chinese herb Pianzihuang (片仔癀) in human osteosarcoma U-2OS cells[J].zhongguo gu shang / China J Orthop Trauma ,2009,22(4):265~268 |
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