中药片仔癀胶囊对骨肉瘤U-2OS细胞诱导凋亡的作用  

Apoptosis induction of traditional Chinese herb Pianzihuang (片仔癀) in human osteosarcoma U-2OS cells

作　　者：张俐[1] 于波[1] 林建华[2] 

机构地区：[1]福建中医学院骨伤系,福建福州350108 [2]福建医科大学附属第一医院

出　　处：《中国骨伤》2009年第4期265-268,共4页China Journal of Orthopaedics and Traumatology

基　　金：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]

摘　　要：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120g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100IU/ml,链霉素100μg/ml的RPMI1640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。④TUNEL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:①20%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。③20%片仔癀胶囊含药血清组TUNEL检测阳性。④DNALadder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。To study the effect of apoptosis induction of Pionzihuang 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 Pian.zihuang. ②Human osteosarcoma U-2OS ceils 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 morphons of osteosarcoma U-2OS cell were observed by inverted phase contrast microscope and electron mi- croscope. 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 pro- liferation 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 concen- tration 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 “adder” 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 ：Piaztzihuan,e can inhibit oroliferation and induce aooptosis for osteosarcoma U-2OS cell

关 键 词：细胞凋亡 中药 片仔癀 骨肉瘤U-2OS细胞 

分 类 号：R738.1[医药卫生—肿瘤] R282.71[医药卫生—临床医学]