摘要
目的探讨生物碱类中药成分小檗碱(berber-ine)诱导人肺癌GLC-82细胞的凋亡作用。方法采用细胞培养技术,用不同浓度的小檗碱处理GLC-82细胞48h,用Hoechst33258染细胞核,在荧光显微镜下观察凋亡细胞核的形态变化,用QWin图象分析软件测量细胞核面积,计算凋亡率。采用MTT法测定细胞的增殖能力。小檗碱作用GLC-82细胞36h后,采用荧光免疫细胞化学SABC-Cy3法测定Caspase-3和PARP蛋白表达,在荧光显微镜下拍照,用QWin图象分析软件测定Caspase-3和PARP荧光强度值。结果小檗碱剂量依赖性的抑制GLC-82细胞增殖,半数生长抑制剂量IC50为19.9mg·L-1,上调凋亡促进因子Caspase-3,PARP蛋白表达增加,使GLC-82细胞呈现典型的凋亡形态学特征,胞核缩小,核质浓集,呈斑块状,有凋亡小体。结论小檗碱通过上调Caspase-3诱导人肺癌GLC-82细胞凋亡。
AIM： To study the effect of berberine, alkaloid, on GLC-82 cell apoptosis. METHODS： GLC- 82 cells were cultured with different concentrations of berberine. Proliferation inhibition of berberine on GLC-82 cells was evaluated by MTF assay. Morphologic changes of berberine-treated cells were observed by light microscope. Apoptotic cells with condensed or fragmented nuclei were visualized by Hoechst33258 by a fluorescence microscope. Nuclei area and apoptotic rate were measured with the use of QWin. The protein expressions of Caspase-3 and PARP were analyzed by fluorescence immunohistochemical method （SABC-Cy3）, fluorescent intensity of Caspase-3 and PARP were measured with the use of QWin. RESULTS： 48 h after berberine treatment, apoptotic cells increased dose-dependently, morphological changes of GLG-82 Ceils showed many hallmark features of apoptosis, including nuclei dwindle and chromatin aggregation and fragmented nuclei. Inhibition of GLC-82 cell proliferation was concentration-dependent, the IC50 value for berberine was 19.9 mg·L^-1. 36 h after berberine treatment, the measurement of fluorescent intensity showed 20 mg·L^-1 of berberine increased the intensity levels of Caspase-3 and PARP. CONCLUSION： The berberine evoked GLC-82 cell apoptosis is correlation with the up-regulation of Caspase-3. This study indicated that berberine might be a potential Chinese medical component for lung neoplasm.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2005年第12期1389-1393,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
国家自然科学基金资助项目(№30200235)
