癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2014年
6期
401-407
,共7页
李登科%李宝赛%崔宝弟%孙震晓
李登科%李寶賽%崔寶弟%孫震曉
리등과%리보새%최보제%손진효
何首乌%大黄素-8-O-β-D-葡萄糖苷%抗癌活性%细胞周期%细胞凋亡
何首烏%大黃素-8-O-β-D-葡萄糖苷%抗癌活性%細胞週期%細胞凋亡
하수오%대황소-8-O-β-D-포도당감%항암활성%세포주기%세포조망
Polygonum multiflorum Thunb%emodin-8-O-β-D-glucopyranoside%antitumor activity%cell cycle%apoptosis
目的：分离提取何首乌R50组分中的抗癌活性成分并进一步研究其抗癌作用机制。方法：利用Sephadex LH-20、TLC色谱法、反向硅胶色谱分离系统对何首乌R50组分进行分离纯化，获得大黄素-8-O-β-D-葡萄糖苷，利用TLC和高效液相色谱法(HPLC)鉴定其纯度；MTT法检测大黄素-8-O-β-D-葡萄糖苷对人肝癌细胞HepG2、永生化人肝实质细胞L02、人结直肠癌细胞HT29和HCT116、人神经母细胞瘤细胞SH-SY5Y和人肺癌细胞A549的细胞毒作用；Giemsa染色观察药物作用后HepG2细胞和L02细胞的形态变化；流式细胞术检测大黄素-8-O-β-D-葡萄糖苷对HepG2细胞和L02细胞周期和凋亡的影响。结果：从何首乌R50组分获得大黄素-8-O-β-D-葡萄糖苷，纯度为96%；大黄素-8-O-β-D-葡萄糖苷对HepG2、HT29和SH-SY5Y细胞均有较显著的细胞毒活性(P<0.05)，且对HepG2、HT29细胞的作用表现出剂量效应(r=0.987，=0.002；=0.992，=0.008)，而对L02、HCT116、A549细胞无明显的细胞毒作用；200μg/mL大黄素-8-O-β-D-葡萄糖苷作用HepG2细胞72 h，细胞生长受到抑制，并出现细胞核碎裂等凋亡细胞特征，L02细胞形态正常；大黄素-8-O-β-D-葡萄糖苷可诱导HepG2细胞发生G2~M期阻滞及凋亡。结论：大黄素-8-O-β-D-葡萄糖苷是何首乌R50组分中具抗癌活性的成分之一，其对永生化人肝实质细胞低毒，对人肝癌细胞有显著抑制作用，其作用机制与阻滞癌细胞周期和诱导细胞凋亡有关。
目的：分離提取何首烏R50組分中的抗癌活性成分併進一步研究其抗癌作用機製。方法：利用Sephadex LH-20、TLC色譜法、反嚮硅膠色譜分離繫統對何首烏R50組分進行分離純化，穫得大黃素-8-O-β-D-葡萄糖苷，利用TLC和高效液相色譜法(HPLC)鑒定其純度；MTT法檢測大黃素-8-O-β-D-葡萄糖苷對人肝癌細胞HepG2、永生化人肝實質細胞L02、人結直腸癌細胞HT29和HCT116、人神經母細胞瘤細胞SH-SY5Y和人肺癌細胞A549的細胞毒作用；Giemsa染色觀察藥物作用後HepG2細胞和L02細胞的形態變化；流式細胞術檢測大黃素-8-O-β-D-葡萄糖苷對HepG2細胞和L02細胞週期和凋亡的影響。結果：從何首烏R50組分穫得大黃素-8-O-β-D-葡萄糖苷，純度為96%；大黃素-8-O-β-D-葡萄糖苷對HepG2、HT29和SH-SY5Y細胞均有較顯著的細胞毒活性(P<0.05)，且對HepG2、HT29細胞的作用錶現齣劑量效應(r=0.987，=0.002；=0.992，=0.008)，而對L02、HCT116、A549細胞無明顯的細胞毒作用；200μg/mL大黃素-8-O-β-D-葡萄糖苷作用HepG2細胞72 h，細胞生長受到抑製，併齣現細胞覈碎裂等凋亡細胞特徵，L02細胞形態正常；大黃素-8-O-β-D-葡萄糖苷可誘導HepG2細胞髮生G2~M期阻滯及凋亡。結論：大黃素-8-O-β-D-葡萄糖苷是何首烏R50組分中具抗癌活性的成分之一，其對永生化人肝實質細胞低毒，對人肝癌細胞有顯著抑製作用，其作用機製與阻滯癌細胞週期和誘導細胞凋亡有關。
목적：분리제취하수오R50조분중적항암활성성분병진일보연구기항암작용궤제。방법：이용Sephadex LH-20、TLC색보법、반향규효색보분리계통대하수오R50조분진행분리순화，획득대황소-8-O-β-D-포도당감，이용TLC화고효액상색보법(HPLC)감정기순도；MTT법검측대황소-8-O-β-D-포도당감대인간암세포HepG2、영생화인간실질세포L02、인결직장암세포HT29화HCT116、인신경모세포류세포SH-SY5Y화인폐암세포A549적세포독작용；Giemsa염색관찰약물작용후HepG2세포화L02세포적형태변화；류식세포술검측대황소-8-O-β-D-포도당감대HepG2세포화L02세포주기화조망적영향。결과：종하수오R50조분획득대황소-8-O-β-D-포도당감，순도위96%；대황소-8-O-β-D-포도당감대HepG2、HT29화SH-SY5Y세포균유교현저적세포독활성(P<0.05)，차대HepG2、HT29세포적작용표현출제량효응(r=0.987，=0.002；=0.992，=0.008)，이대L02、HCT116、A549세포무명현적세포독작용；200μg/mL대황소-8-O-β-D-포도당감작용HepG2세포72 h，세포생장수도억제，병출현세포핵쇄렬등조망세포특정，L02세포형태정상；대황소-8-O-β-D-포도당감가유도HepG2세포발생G2~M기조체급조망。결론：대황소-8-O-β-D-포도당감시하수오R50조분중구항암활성적성분지일，기대영생화인간실질세포저독，대인간암세포유현저억제작용，기작용궤제여조체암세포주기화유도세포조망유관。
OBJECTIVE:Polygonum multiflorum Thunb(P. multiflorum) has been used in the treatment of diarrhea and detoxification in China. Our previous studies showed that R50 part which was separated from P. multiflorum had little toxicity to normal cells but certain toxicity to cancer cells. This study aimed to separate the component which has different effect on normal and cancer cells in R50 part and investigate its anticancer mechanisms. METHODS:Sephadex LH-20,TLC chromatography and reverse silica gel chromatography were employed to separate and purify the emodin-8-O-β-D-glucopyranoside(PMEG). The purity was evaluated by TLC and HPLC. The viability of cells treated with PMEG at different concentration and time was detected by MTT assay. Morphological changes in cells and nuclei were examined after Giemsa staining. Cell cycle and apoptosis were assayed by flow cytometry. RESULTS:The purity of PMEG separated from P. multiflorum was 96%. MTT method showed that PMEG had significant growth inhibitory effects on HepG2,HT29,and SH-SY5Y cells(P<0.05),and showed concentration dependence in HepG2 cells and HT29 cells(r=0.987,P=0.002;r=0.992,P=0.008). But PMEG had no effects on L02、HCT116 and A549 cells. PMEG (200 μg/mL) showed much more growth inhibition in HepG2 cells to L02 cells,and induced HepG2 cells apoptosis and G2-M phase block. CONCLUSION:PMEG was one of the main active anticancer component separated from R50 part of P. multiflorum,and had different effects on human liver L02 cells and liver cancer HepG2 cells,which may be related to PMEG-induced apoptosis and G2-M block in HepG2 cells.
