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[摘要]
目的研究冬凌草甲素注射剂诱导人胃癌SGC-7901细胞凋亡及与细胞内Ca2+浓度([Ca2+]i)的关系。方法 MTT法检测冬凌草甲素注射剂对SGC-7901细胞的细胞毒活性;倒置显微镜观察冬凌草甲素注射剂对细胞形态学的影响;流式细胞术测定细胞凋亡率;激光共聚焦显微术检测SGC-7901细胞[Ca2+]i的变化;JC-1单染,激光共聚焦显微术检测SGC-7901细胞中线粒体膜电位的变化。结果冬凌草甲素注射剂对SGC-7901细胞的IC50为21.74μmol/L;作用48 h后,细胞生长密度变疏,细胞皱缩,其中冬凌草甲素注射剂高浓度组大部分细胞破碎。冬凌草甲素注射剂5.5、11、22μmol/L作用24 h后SGC-7901细胞凋亡率分别为7.07%、18.57%、22.96%;钙离子荧光强度高于对照组;线粒体膜电位荧光强度显著低于对照组。结论冬凌草甲素注射剂通过升高SGC-7901细胞[Ca2+]i诱导细胞凋亡。
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[Abstract]
Objective To study the apoptosis of Oridonin Injection against human gastric carcinoma SGC-7901 cell line and its relationship with intracellular calcium concentration([Ca2+]i ). Methods The cytotoxicy of Oridonin Injection against SGC-7901 cells was evaluated by MTT assay. Morphology of SGC-7901 cells was observed by inverted microscope. Cell apoptosis rate was measured by flow cytometry. [Ca2+]i of SGC-7901 cells and mitochondrial membrane potential (MMP)were determined by laser scanning confocal microscope. Results IC50 of Oridonin Injection to SGC-7901 cells was 21.74 μmol/L. Cell density decrease, cell shrinkage, and cell breakage in high-dose group were observed 48 h after administration. The apoptosis rate values of 5.5, 11, and 22 μmol/L Oridomin Injection groups 24 h after administration were 7.07%, 18.57%, and 22.96%, respectively. Fluorescence intensity (FI) values of [Ca2+]i were significantly higher than those in control group. And FI values of MMP were significantly lower than those in control group. Conclusion Oridonin Injection could induce cell apoptosis by increasing [Ca2+]i of SGC-7901 cells.
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[基金项目]
黑龙江省新世纪优秀人才支持计划(1251-NCET-019);黑龙江省研究生创新基金项目(YJSCX2011-163HLJ)