CAJ | 학술논문

目的研究重楼皂苷Ⅰ (PS Ⅰ)与重楼皂苷Ⅱ(PSⅡ)对胶质瘤细胞U251的体外抑制作用及其机制. 方法体外常规培养胶质瘤细胞株U251.用PS Ⅰ与PSⅡ分别处理胶质瘤细胞株U251,并设空白对照组.噻唑蓝(MTT)法测定PS Ⅰ与PSⅡ对胶质瘤细胞株U251的体外抑制作用;Hoechst33258荧光染色检测细胞核发生凋亡的形态变化;采用流式细胞术(FCM)-AnnexinV/PⅠ双染法检测细胞凋亡变化;Western blotting检测Fas、Caspase-8和Caspase-3蛋白表达情况. 结果 MTT分析表明PS Ⅰ与PSⅡ均能抑制U251细胞增殖,并呈时间和剂量依赖性;与空白对照组比较,PS Ⅰ与PSⅡ处理组对U251增殖抑制明显增强,差异有统计学意义(P＜0.05);PSⅠ对胶质瘤细胞U251的抑制作用强,PSⅡ次之.Hoechst33258荧光染色显示典型的凋亡特征.经PS Ⅰ与PSⅡ处理后U251细胞的早期凋亡率明显高于空白对照组(F=81.434,p=0.000).与空白对照组比较,PSⅠ与PSⅡ处理组Fas、Caspase-8和Caspase-3表达上调,差异有统计学意义(P＜0.05). 结论 PSⅠ与PSⅡ对胶质瘤细胞均有明显的抗肿瘤作用,其作用机制与上调Fas、Caspase-8和Caspase-3表达促进细胞凋亡有关.
목적 연구중루조감Ⅰ (PS Ⅰ)여중루조감Ⅱ(PSⅡ)대효질류세포U251적체외억제작용급기궤제. 방법 체외상규배양효질류세포주U251.용PS Ⅰ여PSⅡ분별처리효질류세포주U251,병설공백대조조.새서람(MTT)법측정PS Ⅰ여PSⅡ대효질류세포주U251적체외억제작용;Hoechst33258형광염색검측세포핵발생조망적형태변화;채용류식세포술(FCM)-AnnexinV/PⅠ쌍염법검측세포조망변화;Western blotting검측Fas、Caspase-8화Caspase-3단백표체정황. 결과 MTT분석표명PS Ⅰ여PSⅡ균능억제U251세포증식,병정시간화제량의뢰성;여공백대조조비교,PS Ⅰ여PSⅡ처리조대U251증식억제명현증강,차이유통계학의의(P＜0.05);PSⅠ대효질류세포U251적억제작용강,PSⅡ차지.Hoechst33258형광염색현시전형적조망특정.경PS Ⅰ여PSⅡ처리후U251세포적조기조망솔명현고우공백대조조(F=81.434,p=0.000).여공백대조조비교,PSⅠ여PSⅡ처리조Fas、Caspase-8화Caspase-3표체상조,차이유통계학의의(P＜0.05). 결론 PSⅠ여PSⅡ대효질류세포균유명현적항종류작용,기작용궤제여상조Fas、Caspase-8화Caspase-3표체촉진세포조망유관.
Objective To investigate the inhibitory effects ofPolyphyllin D (Paris saponin [PS]Ⅰ) and formosanin C (PS Ⅱ) on human glioblastoma U251 cells and its mechanism.Methods Glioma cell line U251 was cultured in vitro.Different treatments (PSⅠ and PS Ⅱ) were added into the medium cultured human malignant glioma U251 cells; and blank control group was also established.The effects of PSⅠ and PS Ⅱ on proliferation of glioma cells U251 in vitro was examined using methyl thiazolyl tetrazolium (MTT) assay.The nuclear morphology changes of apoptotic cells were detected by Hoechst33258 fluorescent staining.Apoptotic rate was quantified by flow cytometry (FCM) using Annexin-V/PⅠ dual staining.Western blotting was used to evaluate the changes of Fas,Caspase-8 and Caspase-3 proteins in U251 cells.Results PS Ⅰ and PS Ⅱ inhibted U251 cell proliferation with a timeand dose-dependent manner; as compared with that in the blank control group,the proliferation rate in the PSⅠ and PS Ⅱ treatment groups was significantly increased (P＜0.05); PS Ⅰ had stronger inhibited effect than PS Ⅱ.Typical morphological changes of apoptosis were observed in U251 cells of the PSⅠ and PS Ⅱ treatment groups with Hoechst 33258 staining.The early apoptotic rates of the PSⅠ and PS Ⅱ treatment groups were all significantly higher than that of the blank control groups (F=81.434,P=0.000).As compared with those in the blank control group,the expressions ofFas,Caspase-8 and Caspase-3 were significantly increased (P＜0.05).Conclusion PSⅠ and PS Ⅱ inhibite the proliferation ofglioma cells in vitro; they increase the expressions ofFas,Caspase-8 and Caspase-3,and accelerate the cell apoptosis,which might be the mechanism of inhibition.