中文摘要：

目的：存在于细胞表面的糖复合物参与细胞的通讯、增殖、迁移、分化等生理过程,在机体中具有复杂的生物学调控作用。探讨氮杂糖化合物SZL在体外对人宫颈癌HeLa细胞生长、DNA损伤、线粒体膜电位以及细胞凋亡的干预。方法：实验于2004-12/2006-12在北京大学医学部细胞生物学系完成。①实验材料：人子宫颈癌HeLa细胞株由北京大学医学部细胞生物学系杜晓娟副教授提供。SZL由北京大学天然药物及仿生药物重点实验室叶新山教授合成。②实验方法：取对数生长期HeLa细胞,消化后制成细胞悬液,按1.5×10^3/孔的密度接种,培养24h细胞完全贴壁后,SZL组加入5,10,25,50,100μmol/L的SZL,空白对照组加入DMEM培养液。③实验评估：SZL作用24,48,72h后,采用酸性磷酸酶法检测细胞生长抑制情况,瑞氏染色镜下观察细胞形态;AnnexinV-PI染色后,流式细胞仪检测细胞凋亡率;Rhodamine123标记活细胞后,流式细胞仪检测线粒体膜电位的变化;WesternBlot法检测SZL作用后凋亡相关蛋白的表达;单细胞凝胶电泳检测SZL作用后细胞DNA损伤情况。结果：①细胞生长抑制：5,10,25,50,100μmol/LSZL作用24,48,72h后的半数抑制浓度分别为73.6,43.2,33.6μmol/L,显著抑制Hela细胞的增殖。镜下空白对照组HeLa细胞分布均匀,胞质透明清亮,呈铺展状态生长;50μmol/LSZL作用24h后,HeLa细胞密度变稀,体积变小,胞核染色质呈聚集状态。②细胞凋亡：50,100μmol/LSZL作用24h后细胞凋亡率分别为（3.51±0.41）%,（58.46±8.45）%;在24～48h过程中,凋亡的细胞逐渐坏死,SZL作用48h后细胞凋亡率分别为（10.51±2.20）%,（17.29±7.52）%。空白对照组24,48h后细胞凋亡率均为1%。③线粒体跨膜电位的变化：50μmol/LSZL作用24,48h后,HeLa细胞的线粒体膜电位平均荧光强度均明显低于空白对照组（P〈0.01）。④凋亡相关蛋白的表达：25,50,100μmol/LSZL作用HeLa细胞48h后,凋亡?

英文摘要：

AIM： Carbohydrate complexes on the surface of cells participate the communication, proliferation, migration, and differentiation of cells, and therefore play the biological regulation role in body. In this study, the effects of iminosugar derivative SZL on the proliferation of human cervical carcinoma HeLa cells, DNA damage, mitochondrial trans-membrane potential and cell apoptosis were investigated. METHODS： The experiment was performed in Department of Cell Biology of Peking University Health Science Center from December 2004 to December 2006. （1）Trypsinized human cervical carcinoma cell line HeLa （provided by Professor Du, Department of Cell Biology of Peking University Health Science Center） at log phase were plated culture plates with 1.5×10^3 cells in each hole. When the cells completely attached after 24 hours, SZL groups were given 5, 10, 25, 50, and 100 μmol/L SZL （synthesized by Professor Ye, State Key Laboratory of Natural and Biomimetic Drugs, Peking University）, respectively, while the control group was added by DMEM medium solution. （2）The acid phosphatase assay （APA） was used to determine cell viability after cells were cultured for 24, 48 and 72 hours in SZL. Cell appearance was observed under Wright staining; Cell apoptosis was assayed by flow cytometry after Annexin V /PI double staining. Rhodamine123 staining was used to detect mitochondrial trans-membrane potentials. The expression of apoptosis-related proteins was evaluated by Western Blot. DNA damage was detected by single cell gel electrophoresis. RESULTS： （1）lnhibition effect of SZL in HeLa cells： SZL （5, 10, 25, 50, and 100 μmol/L） inhibited the proliferation of HeLa cells significantly, with 50% inhibiting concentration （IC50） values of 73.6, 43.2, and 33.6 μmol/L at 24, 48, and 72 hours, separately. Under the microscope, the HeLa cells in control group distributed evenly with clear kytoplasm growing spreadingly; after 50 μmol/L SZL treatment for 24 hours, the cells became scarce, and small wi