二甲基氨氯吡咪诱导HL-60细胞分化(英文)

目的:研究阿米洛利衍生物二甲基阿米洛利(DMA)对培养的HL-60细胞增殖和分化的影响。方法:MTT法测定细胞增殖,细胞染色和NBT还原试验测定细胞分化。结果:DMA以浓度依赖方式抑制HL-60细胞增殖,药物作用96小时测得半数抑制浓度(IC_(50))为31.7(95％可信限为6.3-57.1)μmol·L~(-1)。DMA诱导HL-60细胞向粒细胞系分化。加入DMA100μmol·L~(-1)作用3天,细胞分化率从6.5％升至70％。阿米洛利及阿米洛利的另二个衍生物对细胞分化无明显作用。结论:二甲基阿米洛利抑制HL-60细胞增殖和诱导HL-60细胞分化。     

线粒体是双乙酰丙酮氧钒在肾上皮细胞中诱导产生活性氧物种的主要来源(英文)

本研究利用具有降糖效应的双乙酰丙酮氧钒确定其在两种肾上皮细胞系LLC-PK1和MDCK中诱导产生活性氧物种的来源。利用四种常用的荧光试剂分别检测了VO(acac)2诱导产生的过氧化氢(H2O2)和超氧阴离子(·O2)的水平并确定了它们在细胞内的主要来源部位。实验结果表明,VO(acac)2在LLC-PK1和MDCK两种肾细胞系中均能显著诱导ROS的生成,并且ROS主要来源于线粒体。本研究结果提示,可通过局部降低线粒体部位ROS的水平来减少钒化合物的毒性损伤,同时不影响钒化合物的活性。     

槲皮素诱导人白血病HL-60细胞凋亡(英文)

目的:研究槲皮素是否能诱导人白血病HL-60细胞凋亡.方法:应用琼脂糖凝胶电泳法观察DNA碎片;采用流式细胞仪检测DNA断裂;电镜技术观察凋亡的形态学改变,用MTT测定法测定细胞增殖.结果:槲皮素15-120 μmol·L~(-1)诱导HL-60细胞凋亡,电镜观察到典型的形态学改变,电泳显示梯状条带,槲皮素能剂量依赖性地触发DNA降解及抑制细胞增生(IG_(50)和95％可信区间分别为43(30-61)μmol·L~(-1).结论:槲皮素诱导人白血病HL-60细胞凋亡.     

氯化钆对人脐静脉内皮细胞体外血管形成能力的影响(英文)

钆离子的配合物作为核磁共振造影增强剂在临床诊断中得到广泛应用。晚期肾病患者多次大剂量使用钆造影剂后,可能发生肾源性系统纤维化,目前其确切机理还不清楚。我们考察了不同浓度的氯化钆对人脐静脉内皮细胞的迁移和成管能力的影响;在鸡胚尿囊膜模型中,10μM氯化钆诱导生成更多的新生血管。这些实验结果表明氯化钆具有促进人脐静脉内皮细胞体外血管形成能力,这种作用涉及细胞内活性氧物种和钙离子水平的变化,以及PKCα/β2和MAPKs信号通路的激活。该研究为探索钆造影剂和肾源性系统纤维化之间的关系提供了新线索。     

Nrf2活化正向调控紫草素诱导的HL-60细胞分化

目的探讨核因子E2相关因子2(Nrf2)在紫草素诱导HL-60细胞分化中的作用。方法取对数生长期HL-60细胞,分别采用不同浓度紫草素、Nrf2激动剂叔丁基对苯二酚(t BHQ)、Nrf2抑制剂鸦胆子苦醇作用。通过Hoechst 33258染色、MTT法观察HL-60细胞形态和增殖情况,NBT试验检测HL-60细胞还原能力,流式细胞术检测HL-60细胞分化率。ELISA法测定HL-60细胞核内Nrf2含量,RT-PCR或real-time PCR法检测HL-60细胞Nrf2、CD11b、CD14 m RNA表达水平及Nrf2下游基因的表达水平。结果经紫草素(50~200μg·L-1)作用,HL-60细胞形态、还原能力均趋向成熟,与空白对照相比细胞抑制率、分化率、Nrf2含量及其下游基因GCLC、NQO1、GCLM m RNA的表达量均升高(P<0.05或P<0.01),且随紫草素浓度增高有上升的趋势。紫草素与t BHQ联合作用,HL-60细胞CD11b、CD14、NQO1、GCLM m RNA表达量高于紫草素组(P<0.05或P<0.01);紫草素与鸦胆子苦醇联合作用,HL-60细胞CD11b、CD14、GCLM m RNA表达量低于紫草素组(P<0.05或P<0.01)。结论 Nrf2活化正向调控紫草素诱导的HL-60细胞分化。     

HPLC-UV法测定广东王不留行中槲皮素含量(英文)

本文首次建立了广东王不留行活性成分水解后槲皮素的含量测定方法,为评价广东王不留行的质量和制定其质量标准提供依据。采用HPLC-UV法,以C18反相色谱柱,甲醇–0.4%(v/v)磷酸溶液(50:50,v/v)为流动相等度洗脱,检测波长为360nm。对11批广东王不留行样品进行测定,实验证明该含量测定方法分离效果好、灵敏度高、重复性好,平均回收率为99%,可作为广东王不留行的质量控制方法。     

柚皮苷的人肠内菌生物转化(英文)

柚皮苷(1)是酸橙中含量最高的二氢黄酮苷类化合物, 将其与人肠内菌丛共温孵, 从温孵物中通过色谱方法得到原形化合物1和四个转化产物 (2-5), 根据谱学数据确定它们分别为柚皮苷-6"-乙酰物(2)、柚皮素(3)、根皮酸(4)和间苯三酚(5)。在1的糖苷链葡萄糖基6位能够特异性地发生乙酰化作用。1口服难以吸收。本文结果为阐明其生物利用度低提供了实验依据, 推测表现其生物活性的物质基础主要是其肠内菌生物转化产物3。肠内菌丛所致1的生物转化的继发结果,是它的转化产物吸收进入体循环发挥生物学作用, 此结果为进一步或再评价其体内过程、作用或毒性及其生物利用度提供了有用的信息。     

大豆甙元(S86019)与乳香有效成分Bc-4或阿糖胞苷对HL-60细胞分化的联合诱导

大豆甙元($86019)单独处理HL-60细胞,对细胞生长诱导分化作用较弱,NBT阳性细胞数低于10%。当S86019与乳香有效成分Bc-4联合应用时,对HL-60细胞的生长有明显抑制,并有明显的分化诱导;S86019与Bc-4或Ara-c联合应用,对细胞的增殖有较强的抑制和分化诱导作用。S86019与Bc-4联合应用4d,NBT阳性细胞达80%,75%细胞获得吞噬乳胶颗粒能力,90%细胞向成熟粒细胞方向分化;S86019与Ara-c联合应用4d,NBT阳性反应细胞达70%,82%细胞获得吞噬能力,90%细胞形态发生分化。S86019与Bc-4或Ara-c联合用药4d,可明显阻断细胞由G1期向s期移行,G1期细胞数明显增加。     

环孢菌素抑制钙离子介导的HL-60细胞凋亡(英文)

目的:研究环孢菌素(Cyc)在HL-60细胞凋亡中的作用.方法:通过DNA凝胶电泳、细胞形态观察及流式细胞术等方法确定药物三尖杉酯碱(Har), 喜树碱(Cam)或卡西霉素(Cal),thapsigargin(Tha)诱导的细胞凋亡.流式细胞术测定细胞凋亡中胞内相对自由钙浓度的变化.结果:Cal或Tha均能诱导HL-60细胞凋亡,其效应可被Cyc抑制;但Cyc不抑制Hat或Cam诱导的HL-60细胞凋亡.Cal或Tha均能诱导细胞内Ca~(2 )浓度升高,而Hat或Cam并不诱导胞内Ca~(2 )浓度升高.结论:Cyc只抑制由Ca~(2 )升高介导的细胞凋亡.Cal或Tha诱导的细胞凋亡机制不同于由Hat或Cam所诱导的细胞凋亡机制.     

《中国药学》(英文版)2013年致谢

中国药学英文版, 致谢     

Glaucocalyxin A induces apoptosis in human leukemia HL-60 cells through mitochondria-mediated death pathway

Glaucocalyxin A (GLA) is a biologically active ent-kauranoid diterpenoid isolated from Rabdosia japonica var. glaucocalyx, a traditional Chinese medicinal herb, which has been shown to inhibit tumor cell proliferation. However, the mechanism underlying GLA-induced cytotoxicity remains unclear. In this study, we focused on the effect of GLA induction on apoptosis, the mitochondria-mediated death pathway and the accumulation of reactive oxygen species (ROS) in human leukemia cells (HL-60). GLA could induce a dose-dependent apoptosis in HL-60 cells as characterized by cell morphology, DNA fragmentation, activation of caspase-3, -9 and an increased expression ratio of Bax/Bcl-2. The mitochondrial membrane potential (Δψ_m) loss and cytochrome c release from mitochondria to cytosol were observed during the induction. Moreover, GLA caused a time- and dose-dependent elevation of intracellular ROS level in HL-60 cells, and N-acetyl-l-cysteine (NAC, a well-known antioxidant) could block GLA-induced ROS generation and apoptosis. These data suggest that GLA induces apoptosis in HL-60 cells through ROS-dependent mitochondrial dysfunction pathway.     

氢醌抑制HL-60细胞分化上调过氧化物酶2-Cys过氧化物氧还蛋白表达

目的研究氢醌(HQ)对HL-60细胞向单核系、粒系分化的影响。方法 HQ 1,5和50μmo·lL-1分别与豆蔻酰佛波醇乙酯(PMA)20 nmol·L-1或1.25%DMSO共同处理细胞,分别于48和96 h收集细胞。通过观察细胞形态、硝基四氮唑蓝还原反应鉴定细胞分化;CCK-8检测细胞增殖,荧光定量PCR检测2-Cys过氧化物氧还蛋白(Prxs)基因表达的变化;Western印迹检测2-Cys Prxs蛋白表达的变化。结果在HQ1~50μmol·L-1作用下,HL-60细胞向单核系分化受到抑制;HQ1和5μmol·L-1对DMSO诱导的粒系分化无影响,但HQ50μmol·L-1可抑制细胞向粒系分化。HQ5和50μmol·L-1与诱导剂的共同作用可以抑制HL-60细胞增殖。与正常对照组比较,PMA和DMSO组2-Cys Prxs基因表达水平均有降低的趋势,HQ1,5和50μmol.L-1+PMA20 nmol·L-1组PrxⅠ,PrxⅢ和PrxⅣ各个基因表达水平与PMA组比较均有增高的趋势;DMSO诱导分化组中仅HQ 50μmol·L-1+1.25%DMSO组PrxⅠ和PrxⅢ基因表达水平与单独DMSO组比较显著增高(P<0.05)。结论 HQ1和5μmol.L-1可以抑制HL-60细胞向单核系分化,HQ50μmol·L-1可抑制细胞向粒系分化,并上调PrxⅠ和PrxⅢ基因的表达。     

Chimaphilin induces apoptosis in human breast cancer MCF-7 cells through a ROS-mediated mitochondrial pathway

Chimaphilin, 2,7-dimethyl-1,4-naphthoquinone, is extracted from pyrola [Passiflora incarnata Fisch.]. In this study, the anticancer activity and underlying mechanisms of chimaphilin toward human breast cancer MCF-7 cells are firstly investigated. Chimaphilin could inhibit the viability of MCF-7 cells in a concentration-dependent manner, and the IC₅₀ value was 43.30 μM for 24 h. Chimaphilin markedly induced apoptosis through the investigation of characteristic apoptotic morphological changes, nuclear DNA fragmentation, annexin V-FITC/propidium iodide (PI) double staining. Flow cytometry assay revealed that chimaphilin triggered a significant generation of ROS and disruption of mitochondrial membrane potential. Additionally, western blotting assay showed that chimaphilin suppressed Bcl-2 level and enhanced Bad level, then activated caspase-9 and caspase-3, and further activated the poly ADP-ribose polymerase (PARP), finally induced cell apoptosis involving the mitochondrial pathway. Furthermore, free radical scavengers N-acetyl-L-cysteine (NAC) pretreatment test testified that chimaphilin could increase the generation of ROS, then induce cell apoptosis. In general, the present results demonstrated that chimaphilin induced apoptosis in human breast cancer MCF-7 cells via a ROS-mediated mitochondrial pathway.     

Costunolide induces differentiation of human leukemia HL-60 cells

Costunolide has been reported to be a cytotoxic and chemopreventive agent. This work investigated the mechanism of the antiproliferative effect of costunolide and determined that it induced differentiation of the human leukemia cell line HL-60. Costunolide exhibited a potent antiproliferative activity against HL-60 cells. It was also found to be a potent inducer of differentiation in human leukemia derived HL-60 cells through the examination of differentiation markers, as assessed by the reduction of nitroblue-tetrazolium, the increase in esterase activities and phagocytic activity, morphology change and the expression of CD14 and CD66b surface antigens. These results, accompanied by a decline in the expression of c-myc protein, suggest that costunolide induces differentiation of human leukemia cells to granulocytes and monocytes/macrophages lineage.     

三种中药皂甙对HL—60细胞诱导分化的实验研究

目的：研究人参总皂甙、绞股蓝总皂甙和三七总皂甙诱导白血病细胞分化及其机理，为进一步开发三种皂甙的临床应用提供理论与实验依据。方法：采用细胞体外培养、流式细胞术、形态学观察、组化与免疫细胞学等方法，研究人参总皂甙（TSPG）、绞股蓝总皂甙（GP）和三七总皂甙（PNS）对HL－60细胞诱导分化的影响。结果：三种中药皂甙均能阻止HL－60细胞由G0／G1期向G2／M＋S期过渡；经三种中药皂甙诱导后，HL－60细胞形态趋向成熟分化，NBT还原反应阳性细胞数和过氧化物酶（POX）染色阳性细胞数增加，CD14、CD15表达阳性细胞数也显著提高,但对照组细胞与实验组细胞非特异性酯酶（NAE）染色均为阴性。结论：提示TSPG、GP、PNS诱导HL－60细胞向粒系分化为主。     

盐酸小檗碱对HL-60细胞增殖与分化的影响

目的　研究盐酸小檗碱对人早幼粒白血病HL 6 0细胞增殖与分化的影响。方法　应用生长曲线测定法、克隆形成实验检测药物对HL 6 0细胞生长抑制作用 ;细胞分化根据细胞形态、硝基蓝四氮唑 (NBT)还原能力、细胞表面标记的改变来决定 ;细胞周期变化用流式细胞仪测定。结果　HL 6 0细胞经小檗碱处理后生长明显受抑 ,呈时间和浓度依赖性。选择 1、2、4、8mg·L-1小檗碱作用于HL 6 0细胞 ,动态观察发现 ,HL 6 0细胞向成熟细胞分化 :细胞核变小 ,核浆比减少 ;NBT还原能力增强 ;CD11b表达升高。细胞周期分析发现 ,小檗碱处理后细胞阻滞于G0 /G1期 ,S期细胞明显减少。结论　盐酸小檗碱可抑制HL 6 0细胞的增殖 ,并诱导HL 6 0细胞向成熟细胞分化。     

Capsaicin potentiates 1,25-dihydoxyvitamin D3- and all-trans retinoic acid-induced differentiation of human promyelocytic leukemia HL-60 cells.

Human promyelocytic leukemia HL-60 cells are differentiated into monocytic or granulocytic lineage when treated with 1,25-dihydroxyvitamin D₃ [1,25-(OH)₂D₃] or all-trans retinoic acid, respectively. In this study, the effect of capsaicin, an active component of the red pepper of the genus Capsocum, on cell differentiation was investigated in a HL-60 cell culture system. Treatment of HL-60 cells with 5–30 μg/ml capsaicin for 72 h inhibited cell proliferation and induced a small increase in cell differentiation. Interestingly, synergistic induction of HL-60 cell differentiation was observed when capsaicin was combined with either 5 nM 1,25-(OH)₂D₃ or 50 nM all-trans retinoic acid. Flow cytometric analysis indicated that combinations of 1,25-(OH)₂D₃ and capsaicin stimulated differentiation predominantly to monocytes whereas combinations of all-trans retinoic acid and capsaicin stimulated differentiation predominantly to granulocytes. Capsaicin enhanced protein kinase C activity in 1,25-(OH)₂D₃- and all-trans retinoic acid-treated HL-60 cells. In addition, inhibitors for protein kinase C [bisindolylmaleimide (GF-109203X), chelerythrine, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7)] and an inhibitor for extracellular signal-regulated kinase [2-(2′-amino-3′-methoxyphenyl)-oxanaphthalen-4-one (PD-098059)] significantly inhibited HL-60 cell differentiation induced by capsaicin in combination with either 1,25-(OH)₂D₃ or all-trans retinoic acid. These results indicate that capsaicin potentiates 1,25-(OH)₂D₃- or all-trans retinoic acid-induced HL-60 cell differentiation and that both protein kinase C and extracellular signal-regulated kinase are involved in the cell differentiation synergistically enhanced by capsaicin.     

Dracorhodin perchlorate induces apoptosis in HL-60 cells

Dracorhodin perchlorate, an anthocyanin red pigment, induces human premyelocytic leukemia HL-60 cell death through apoptotic pathway. Caspase -1, -3, -8, -9, and -10 inhibitors partially reversed the cell death induced by dracorhodin perchlorate. Caspase-3 and -8 were activated followed to the degradation of caspase-3 substrates, inhibitor of caspase-activated DNase (ICAD) and poly-(ADP-ribose) polymerase (PARP). Dracorhodin perchlorate up-regulated the expression ratio of mitochondrial proteins, Bax/Bcl-X_L. The cell death was accompanied with phosphorylation of ERK, JNK and p38 MAPK and partially reduced by MEK inhibitor (PD98059), JNK MAPK inhibitor (SP600125) and p38 MAPK inhibitor (SB 203580). Taken together, dracorhodin perchlorate-induced apoptosis in HL-60 cells via up-regulation of Bax, activation of caspases and ERK/p38/JNK MAPKs.