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目的:探讨糙叶败酱大孔吸附树脂提取物(Patrinia scabra Bunge extract,PsBe)引起S180肉瘤细胞凋亡的相关机制。方法:建立S180腹水瘤动物模型,以环磷酰胺(CTX)0.01 g/kg作为阳性对照,分别给予PsBe 0.5、1.0、2.0 g/kg治疗,透射电镜和荧光显微镜观察肿瘤细胞凋亡的形态学变化,Annexin V-FITC/PI双标记法检测肿瘤细胞早期凋亡的数量变化,激光共聚焦显微镜检测肿瘤细胞DNA、RNA含量、细胞间通讯(GJIC)和细胞[Ca2+]的变化。结果:PsBe可降低肿瘤细胞的DNA、RNA含量,透射电镜和荧光显微镜观察可见肿瘤细胞出现凋亡的组织形态学改变;升高肿瘤细胞凋亡率、降低存活率,与模型组比较差异有显著性(P<0.01),但与CTX组比较,PsBe各剂量组肿瘤细胞活细胞比例升高、坏死率降低(P<0.01),仅PsBe 2.0 g/kg组肿瘤细胞凋亡率升高(P<0.01),即PsBe组肿瘤细胞以凋亡为主,CTX组肿瘤细胞则以坏死为主。同时,PsBe可改善细胞间通讯功能状况,荧光恢复率与模型组比较差异有显著性(P<0.01),可升高肿瘤细胞[Ca2+],与模型组比较差异有显著性(P<0.01)。结论:PsBe具有抗肿瘤、诱导S180细胞凋亡作用,其机制与恢复GJIC和细胞[Ca2+]升高有关。
Objective: To investigate the mechanism of apoptosis of S180 sarcoma cells induced by Patrinia scabra Bunge extract (PsBe). Methods: The animal model of S180 ascites tumor was established. CTX 0.01 g / kg was used as positive control and treated with PsBe 0.5, 1.0 and 2.0 g / kg respectively. The morphological changes of tumor cells were observed by transmission electron microscope and fluorescence microscope The changes of early apoptosis in tumor cells were detected by Annexin V-FITC / PI double labeling method. The changes of DNA and RNA, GJIC and [Ca2 +] in tumor cells were detected by confocal microscopy. Results: PsBe reduced the DNA and RNA content of tumor cells. The morphological changes of tumor cells were observed by transmission electron microscope and fluorescence microscope. The apoptotic rate of tumor cells was decreased and the survival rate was decreased, which was significantly different from the model group (P <0.01). However, compared with CTX group, the percentage of viable cells in tumor cells increased and the necrosis rate decreased (P <0.01) <0.01). That is to say, the tumor cells in PsBe group were predominated by apoptosis, and the tumor cells in CTX group were predominantly necrotic. At the same time, PsBe could improve the function of cell-to-cell communication. Fluorescence recovery rate was significantly different from that of the model group (P <0.01), and increased the [Ca2 +] of tumor cells compared with the model group . Conclusion: PsBe has anti-tumor effect and induces the apoptosis of S180 cells. Its mechanism is related to the recovery of GJIC and the increase of [Ca2 +] in cells.