CAJ | 학술논문

目的：探讨去甲斑蝥素（NCTD）及其五种衍生物抗白血病细胞K562的作用．方法：常规方法复苏、传代培养人慢性粒细胞性白血病K562细胞，待细胞进入对数生长期后用于实验．空白对照组行常规培养，药物处理包括去甲斑蝥素（NCTD）处理组和NCTD衍生物（QJ-2、QJ-3、QJ-13、BH052及BH091）处理组，分别设置各组药物终浓度0．001 mmol／L、0．005 mmol／L、0．05 mmol／L、0．1 mmol／L、0．5 mmol／L、1 mmol／L、5 mmol／L、10 mmol／L 处理，各组于处理24 h、48 h （NCTD）、72 h（NCTD）后收集细胞检测：①倒置相差显微镜观察细胞生长状况；②常规瑞氏染色后光学显微镜下观察细胞形态；③MTT比色法检测不同处理组细胞的活力和增殖能力．结果：①与空白对照组比较，NCTD及其衍生物处理后活细胞减少，并出现抱团生长、胞膜出泡、胞体破碎等现象，且变化程度随浓度升高更加明显；瑞氏染色观察 NCTD 作用后K562细胞，药物处理组细胞胞核染色质出现聚集、溶解、碎裂现象，胞浆出现空泡，空泡量且随NCTD浓度升高而增多；②MTT实验测定各药物对K562细胞的IC50值由大到小分别为QJ-3（1．88 mmol／L）、BH091（1．787 mmol／L）、QJ-13（0．621 mmol／L）、BH052（0．406 mmol／L ）、QJ-2（0．129 mmol／L ）、NCTD （0．083 mmol／L），NCTD对K562细胞的增殖抑制作用最强，且该抑制作用随药物剂量增加和作用时间延长而增强；在NCTD的五种衍生物中 QJ-2的抑制作用最强．结论：①NCTD对K562细胞生长有较强的抑制作用，且呈剂量-时间依赖性；②五种衍生物中QJ-2对K562细胞的作用最强．
목적：탐토거갑반모소（NCTD）급기오충연생물항백혈병세포K562적작용．방법：상규방법복소、전대배양인만성립세포성백혈병K562세포，대세포진입대수생장기후용우실험．공백대조조행상규배양，약물처리포괄거갑반모소（NCTD）처리조화NCTD연생물（QJ-2、QJ-3、QJ-13、BH052급BH091）처리조，분별설치각조약물종농도0．001 mmol／L、0．005 mmol／L、0．05 mmol／L、0．1 mmol／L、0．5 mmol／L、1 mmol／L、5 mmol／L、10 mmol／L 처리，각조우처리24 h、48 h （NCTD）、72 h（NCTD）후수집세포검측：①도치상차현미경관찰세포생장상황；②상규서씨염색후광학현미경하관찰세포형태；③MTT비색법검측불동처리조세포적활력화증식능력．결과：①여공백대조조비교，NCTD급기연생물처리후활세포감소，병출현포단생장、포막출포、포체파쇄등현상，차변화정도수농도승고경가명현；서씨염색관찰 NCTD 작용후K562세포，약물처리조세포포핵염색질출현취집、용해、쇄렬현상，포장출현공포，공포량차수NCTD농도승고이증다；②MTT실험측정각약물대K562세포적IC50치유대도소분별위QJ-3（1．88 mmol／L）、BH091（1．787 mmol／L）、QJ-13（0．621 mmol／L）、BH052（0．406 mmol／L ）、QJ-2（0．129 mmol／L ）、NCTD （0．083 mmol／L），NCTD대K562세포적증식억제작용최강，차해억제작용수약물제량증가화작용시간연장이증강；재NCTD적오충연생물중 QJ-2적억제작용최강．결론：①NCTD대K562세포생장유교강적억제작용，차정제량-시간의뢰성；②오충연생물중QJ-2대K562세포적작용최강．
AIM:To analyze and compare activity of anti-human chronic myelogenous leukemia K562 cells of norcantharidin (NCTD ) and its five derivatives.METHODS:The leukemic K562 cells were recovered and cultured with conventional methods and were used for studying in logarithmic phase.The control group was treated by conventional culture and drug treatment in-cluded NCTD and NCTD derivatives (QJ-2, QJ-3, QJ-13, BH052 and BH091)in a series concentrations from 0.001 mmol/L,0.005 mmol/L,0.05 mmol/L,0.1 mmol/L,0.5 mmol/L, 1 mmol/L,5 mmol/L,10 mmol/L.Then the cells were collected at 24 h,48 h(NCTD),72 h (NCTD)post culture for the follow-ing tests:①Cell growth status were observed with inverted phase contrast microscope to observe;② Cell morphology was observed under an optical microscope with Wright stain;③Using MTT col-orimetry to detect cell activity and proliferation.RESULTS:①Compared with the control group,K562 cells treated by NCTD and its derivatives showed the phenomenon of living cell reducing, dead cells increasing,cells growing close together,vacuolation and cell disruption,which was evident with the increasing of the concentration.It was observed that nuclear chromatin aggregation, dissolution,fragmentation,cytoplasmic vacuoles changes in K562 cells with Wright stain after NCTD and its derivatives treatment.② Based on the experimental results of MTT,the IC50 values of drug on cells were QJ-3 (1.88mmol/L),BH091 (1.787 mmol/L),QJ-13 (0.621 mmol/L),BH052 (0.406 mmol/L),QJ-2 (0.129 mmol/L),NCTD (0.083 mmol/L).It showed that the NCTD had the strongest inhibition on K562 cells and was in dose and time dependant manner.QJ-2 showed the strongest inhibition in NCTD five derivatives.CONCLUSION:①NCTD can inhibit strongly growth of K562 cells in a dose-time-dependent way;②QJ-2 showed the strongest inhibition in NCTD five derivatives.