CAJ | 학술논문

目的：探讨何首乌苷保护大鼠星形胶质细胞免于奥沙利铂诱导凋亡的作用及可能机制。方法原代培养大鼠星形胶质细胞，并利用流式检测所培养细胞胶质纤维酸性蛋白的表达情况。利用10μmol/L奥沙利铂处理胶质细胞24 h建立凋亡模型，加入浓度梯度的何首乌苷预处理胶质细胞24 h后与奥沙利铂共同作用24 h，台盼蓝染色法观察细胞存活。随后通过Annexin V-FITC/PI双染法流式检测各组凋亡率和检测Caspase-3的活性水平。进而利用JC-1染色法流式检测细胞线粒体膜电位，并利用实时定量RT-PCR与免疫印迹检测BAX与BCL2的mRNA水平和蛋白表达量。结果培养的大鼠星形胶质细胞表达胶质纤维酸性蛋白率达95%以上。奥沙利铂处理诱导凋亡24 h后，对照组胶质细胞凋亡率及Caspase-3活性上升并降低线粒体膜电位，加何首乌苷处理后凋亡率及Caspase-3活性下降而线粒体膜电位上升。相对于对照组，加入何首乌苷处理后BAX基因的mRNA及蛋白表达水平降低，BCL2基因的mRNA及蛋白表达水平上升。结论何首乌苷能恢复凋亡相关基因BAX与BCL2的平衡，保护大鼠皮质星形胶质细胞的线粒体膜电位，并使胶质细胞免于奥沙利铂诱导的凋亡作用。
목적：탐토하수오감보호대서성형효질세포면우오사리박유도조망적작용급가능궤제。방법원대배양대서성형효질세포，병이용류식검측소배양세포효질섬유산성단백적표체정황。이용10μmol/L오사리박처리효질세포24 h건립조망모형，가입농도제도적하수오감예처리효질세포24 h후여오사리박공동작용24 h，태반람염색법관찰세포존활。수후통과Annexin V-FITC/PI쌍염법류식검측각조조망솔화검측Caspase-3적활성수평。진이이용JC-1염색법류식검측세포선립체막전위，병이용실시정량RT-PCR여면역인적검측BAX여BCL2적mRNA수평화단백표체량。결과배양적대서성형효질세포표체효질섬유산성단백솔체95%이상。오사리박처리유도조망24 h후，대조조효질세포조망솔급Caspase-3활성상승병강저선립체막전위，가하수오감처리후조망솔급Caspase-3활성하강이선립체막전위상승。상대우대조조，가입하수오감처리후BAX기인적mRNA급단백표체수평강저，BCL2기인적mRNA급단백표체수평상승。결론하수오감능회복조망상관기인BAX여BCL2적평형，보호대서피질성형효질세포적선립체막전위，병사효질세포면우오사리박유도적조망작용。
Objective To investigate the effect of tetrahydroxy stilbene glycoside (TSG) preventing astrocytes from oxaliplatin induced apoptosis, and relate the mechanism. Methods Primary cultured astrocytes were exposed to 10 μmol/L oxaliplatin for 24 h for inducing apoptosis, and the TSG was added into the culture medium before the oxaliplatin of 24 h. The rate of apoptosis was assayed by Annexin V-FITC/PI staining and flow cytometry analysis, and the Caspase-3 activity were detected by color metric assay. The mitochondrial membrane potential was detected by JC-1 staining and flow cytometry analysis. The BAX and BCL2 expressions were measured by Real-time quantitive polymerase chain reaction (qRT-PCR) for mRNA level and detected by Western blot for protein level. Results The apoptosis rate and Caspase-3 activity had increased on astrocytes induced by oxaliplatin, but TSG can prevent such effects. Furthermore, the mitochondrial membrane potential had decreased on astrocytes induced by oxaliplatin, while TSG can reverse such effects. Moreover, the oxaliplatin could induce the expression of BAX and suppress the expression of BCL2 on mRNA level and protein level, while the TSG had reversed those effects. Conclusions TSG could prevent astrocytes from oxaliplatin induced apoptosis and restore the balance between BAX and BCL2 to protect the mitochondrial membrane potential normal.