黄樟素氧化物对去除成纤维细胞生长因子所诱导的血管内皮细胞凋亡及生长的影响

目的:研究黄樟素氧化物对去除成纤维细胞生长因子(FGF)诱导的血管内皮细胞凋亡及生长的影响.方法:光学显微镜观察细胞形态学变化;MTT法测定细胞生长;DNA电泳和荧光显微技术检测DNA断裂;流式细胞术测定细胞周期分布.结果:黄樟素氧化物5-25mg/L处理去除FGF的血管内皮细胞24h,细胞铺展和生长被促进,细胞脱壁和DNA片段化被抑制,黄樟素氧化物10mg/L对细胞周期分布无明显影响.黄樟素氧化物50-100 mg/L促进血管内皮细胞的脱壁和DNA片段化,黄樟素氧化物100mg/L将细胞周期阻断于G_2-M期.结论:黄樟素氧化物在10mg/L时抑制血管内皮细胞凋亡,而在100mg/L时促进其凋亡,该药物对血管内皮细胞生长和凋亡有重要影响.     

海鞘素简易结构类似物GJ7-1和GJ7-2的抗肿瘤活性及分子靶向研究

目的:研究依照特有DNA靶向性的抗肿瘤海洋天然产物海鞘素(Ecteinascidins,ETs)合成的简易结构类似物GJ7-1和GJ7-2的抗肿瘤活性及分子靶向。方法:MTT法检测0.012 5¹.6μmol/L的GJ7-1、GJ7-2作用72 h对10种体外培养的肿瘤细胞的增殖抑制作用;荧光结合竞争法测定0.011.6μmol/L的GJ7-1、GJ7-2作用72 h对10种体外培养的肿瘤细胞的增殖抑制作用;荧光结合竞争法测定0.01¹00μmol/L的GJ7-1、GJ7-2与DNA的结合情况;流式细胞仪检测0.01、0.1、1μmol/L的GJ7-1、GJ7-2对A549细胞周期(作用24、48 h)的影响;Hochest33342染色和AnnexinⅤ/PI双染检测0.01、0.1、1μmol/L的GJ7-1、GJ7-2对A549细胞凋亡(作用24、48、72 h)的影响。结果:GJ7-1和GJ7-2对10种肿瘤细胞均有增殖抑制作用,但无明显肿瘤类型选择性;浓度增加到100μmol/L时也与DNA无明显结合;仅1μmol/L的GJ7-1、GJ7-2对A549细胞周期有一定的影响,但均不能诱导其明显凋亡。结论:GJ7-1和GJ7-2虽有一定的抗肿瘤活性,但较ETs大幅降低,部分原因是其完全丧失了与DNA的结合活性。     

阿魏酸钠对抗Aβ25-35诱导的鼠神经元凋亡作用研究

目的 探讨阿魏酸钠对β淀粉样蛋白(Aβ25-35)通过谷氨酸诱导的鼠皮层神经元凋亡的影响.方法 培养的皮层神经元分别与谷氨酸(20μmol/L)、Aβ25-35(5μmol/L)、Aβ25-35(5μmol/L)+谷氨酸(20μmol/L)孵育,采用Hoechst 33258荧光染色法分析神经细胞凋亡;然后用谷氨酸( 50μmol/L)诱导神经元凋亡,采用荧光染色法和Westem blot观察阿魏酸钠的保护作用.结果 单独应用Aβ25-35(5μmol/L)和单独加入谷氮酸(20μmol/L)引起的皮层神经元凋亡率与对照组无明显差异;Aβ25-35与皮层神经元共同孵育5天,接着用谷氨酸处理24h,细胞凋亡率从正常的9.2%+1.5%增加到43%±8%:阿魏酸钠能够显著降低谷氨酸诱导的神经细胞凋亡百分比至21%±5%,并对抗谷氨酸引起的Bcl-2蛋白表达的降低.结论 阿魏酸钠能够减弱Aβ25-35提高谷氨酸毒性诱导的鼠皮层神经元凋亡.     

缺氧对胎盘滋养层细胞细胞周期和凋亡的影响

目的 观察缺氧时滋养层细胞细胞周期及凋亡变化.方法 体外培养胎盘绒毛膜癌滋养层细胞株(BeWo细胞),二氯化钴(CoCl2)处理模拟化学缺氧.噻唑蓝法测定不同浓度(0、125、250、500μmol/L)CoCl2作用不同时相(6、12、24 h)后细胞活力;流式细胞术检测250 μmol/L CoCl2处理细胞不同时间(6、12、24 h)后细胞周期和凋亡改变.结果 125 μmol/L CoCl2对细胞活力无明显影响(P＞0.05);250μmol/L CoCl2作用时细胞活力呈下降趋势(P＞0.05);500μmol/L CoCl2处理6、12、24 h后细胞活力均有明显下降(P＜0.01).250 μmol/L CoCl2诱导构建滋养层细胞缺氧模型.250μmol/L CoCl2作用12、24 h后,G2/M期细胞比例增多(分别为0.24±0.13和0.31±0.01)(P＜0.05或P＜0.01).250 μmol/L CoCl2作用细胞24 h后细胞凋亡呈增加趋势(P＞0.05).结论 成功构建了滋养层细胞体外缺氧培养模型.缺氧诱导滋养层细胞周期向G2/M期进展,增加细胞凋亡.     

4-[4″-(2″,2″,6″,6″-四甲基-1″-哌啶氮氧自由基)氨基]-4′-去甲表鬼臼毒素对K562/ADM细胞的抑制作用

目的比较4-[4″-(2″,2″,6″,6″-四甲基-1″-哌啶氮氧自由基)氨基]-4′-去甲表鬼臼毒素(GP-7)对多药耐药人慢性粒细胞白血病K562的多柔比星耐药株细胞(K562/ADM细胞)的抑制作用是否优于依托泊苷。方法以依托泊苷和K562细胞为对照,用不同浓度GP-7处理K562/ADM细胞不同时间,MTT比色法测定细胞增殖,流式细胞仪测定细胞周期和细胞凋亡率,普通光学显微镜观察细胞凋亡形态,琼脂糖凝胶电泳观察细胞DNA凋亡性降解。结果8~128mol.L-1GP-7处理48h或64μmol.L-1GP-7处理24~72h,GP-7对K562/ADM细胞的增殖抑制呈剂量依赖性(r=0.947,P<0.05)和时间依赖性(r=0.999,P<0.01)。GP-7及依托泊苷对K562/ADM的IC50分别为(45.9±1.8)及(68.7±4.6)μmol.L-1;64μmol.L-1GP-7作用48h可使G2/M期细胞明显增多,相同情况下依托泊苷则使S期细胞明显增多;GP-7可引起K562/ADM和K562细胞凋亡,但其引起的K562/ADM和K562细胞凋亡率与依托泊苷无明显差异;GP-7可引起K562/ADM和K562细胞典型的凋亡形态学变化和DNA凋亡性降解,但GP-7引起的K562/ADM细胞DNA凋亡性降解弱于K562细胞;128及256μmol.L-1GP-7或依托泊苷处理K562/ADM和K562细胞48h,GP-7诱导DNA凋亡性降解的作用强于依托泊苷,但32和64μmol.L-1时作用则相反。结论GP-7可抑制多药耐药白血病细胞株K562/ADM的增殖,诱导细胞凋亡。GP-7抑制多药耐药白血病细胞株K562/ADM的作用优于依托泊苷。     

7- 二氟甲氧基-5, 4′- 二- 正辛烷氧基金雀异黄素诱导肺癌A549 细胞凋亡

目的研究7二氟甲氧基-5,4＇二正辛烷氧基金雀异黄素（7-difluorom＋hoxyl-5,4＇-di-n-octylgenistein,DFOG）诱导人肺癌A549细胞凋亡作用。方法体外培养A549细胞。二氟甲基化或烷基化得到一系列金雀异黄素衍生物。MTT法测定细胞活力;碘化丙啶（PI）染色流式细胞术（FCM）检测细胞凋亡率;琼脂糖凝胶电泳观察DNA梯形条带;Western blot分析Bcl-2和Bax蛋白表达。结果 9个金雀异黄素衍生物较先导化合物GEN具有更强的抗肿瘤活性。其中DFOG对A549细胞活性显示最强的抑制作用,IC50为3.9μmol·L^-1。DFOG（2.0、4.0和8.0μmol·L^-1）作用48h的细胞凋亡率依次增高。8.0μmol·L^-1DFOG处理A549细胞48h导致典型DNA梯形条带形成。8.0μmol·L1DFOG处理A549细胞6h、12h和24h,Bcl-2蛋白表达水平下降,而Bax蛋白表达上升。结论 DFOG诱导A549细胞凋亡作用与其增高Bax/Bcl-2比值有关。     

咪达唑仑上调miR-382-3p表达抑制IL-1β诱导的软骨细胞炎性因子表达和细胞凋亡的机制

目的 探讨咪达唑仑对白介素1β(IL-1β)诱导的软骨细胞炎性因子表达和细胞凋亡的影响及可能机制.方法 体外培养小鼠软骨细胞ATDC5,不同浓度(1μmol/L、3μmol/L、10μmol/L)的咪达唑仑干预IL-1β诱导的ATDC5细胞24 h、IL-1β诱导转染miR-382-3p模拟物的ATDC5细胞24 h或...     

地昔帕明对皮质酮诱导培养的PC12细胞调亡的拮抗作用(英文)

目的:探讨三环类抗抑郁剂的作用机制.方法:运用DNA电泳法、流式细胞仪法、电镜法检测了皮质酮诱导的PC12细胞凋亡并观察了去甲丙米嗪(DIM)的效应 结果:皮质酮10μmol/L处理5d可显著诱导PC12细胞凋亡,凋亡发生率最高达(28±9)％,DNA电泳图谱则表现出典型的梯状条带,DIM的1和5μmol/L使凋亡发生率明显降低并使梯状条带变浅、减轻,细胞的超微结构也有明显改善.结论:DIM对皮质酮诱导的PC12细胞凋亡有拮抗作用,这可能是其抗抑郁效应的细胞机制之一.     

酒石酸锑钾对肝L-02细胞的抑制和凋亡诱导影响

目的 探讨酒石酸锑钾(PAT)对人胚肝L-02细胞的生长、细胞周期和诱导凋亡的影响.方法 采用改良的噻唑蓝(MTT)比色法检测不同浓度PAT处理24、48、72、96及120 h对L-02细胞的生长抑制效应,用PI单染流式细胞术检测不同浓度PAT处理24 h诱导细胞周期的改变情况,用ArmexinV-FITC/PI双染流式细胞术检测不同浓度PAT作用24 h对细胞凋亡的影响.结果 PAT染毒24、48、72、96及120 h,随PAT染毒剂量的增大,L-02细胞生长抑制率明显升高;浓度200μanol/L PAT作用24 h使细胞发生S期阻滞,≥400 μmol/L PAT使细胞发生G1期阻滞;浓度200μmol/L PAT作用24h诱导细胞凋亡,≥400μmol/L PAT可导致细胞坏死.结论 在一定剂量下,PAT可以显著抑制L-02细胞的生长增殖,低剂量PAT主要引起L-02细胞发生S期阻滞和细胞凋亡,高剂量PAT主要引起L-02细胞发生G1期阻滞和细胞坏死.     

外源性羟自由基诱导血管内皮细胞凋亡及卡托普利的干预作用

目的 观察外源性氧自由基（OFR）衍生的羟自由基（OH^-)诱导培养的VECs凋亡及卡托普利（Captopril,Cap)的干预作用，探讨血管内皮细胞（VECs)凋亡（Apoptosis)与心血管疾病的关系。方法 （1）采用改良的Jaffe法，进行人脐静脉血管内皮细胞（HUVECs)原代、传代培养；（2）取4－6代HUVECs，无血清条件下分别为10^-5、10^-4、10^-3、10^-2、10^-1mol/L羟自由基（OH^-)共孵育，观察细胞存活率、贴壁率、形态学、DNA凝胶电泳、DNA片段率及培养液一氧化氮（NO）含量变化；（3）观察10^-3mol/L OH^-诱导HUVECs凋亡及10^-7、10^-6、10^-5mol/L Cap干预后上述参数的变化。结果 （1）正常培养HUVECs存活率＞95％，贴壁率＞90％；（2）外源性OH－明显降HUVECs存活率、贴壁率，且呈剂量依赖性；10^-5、10^-4、10^-3mol/L OH^-诱导HUVECs凋亡，DNA“梯形”以10^-3mol/L OH^-作用8－24h显著，DNA片段率、凋亡细胞百分率明显升高；而10^-2、10^-1mol/L OH^-可诱导HUVECs坏死；（3）10^-7、10^-6、10^-5mol/L Cap干预明显提高OH^-处理的HUVECs存活率、贴壁率，降低其凋亡细胞百分率、DNA片段率，使DNA“梯形”渐消失，NO分泌减少渐逆转。结论 （1）采用改良的Jaffe法，可以建立HUVECs体外模型；（2）外源性OH^-在较低浓度时可诱导培养HUVECs的典型凋亡；（3）Cap能减轻或抑制OH^-诱导培养HUVECs的凋亡，恢复其NO分泌。     

Effect of manoalide on apoptosis induced by deprivation of growth factors in vascular endothelial cells.

AIM: To study effect of manoalide on apoptosis induced by deprivation of acidic fibroblast growth factor (aFGF) and serum in vascular endothelial cells (VEC). METHODS: Morphologic changes were observed by light microscopy. Viability was determined by counting the cells that attached to dishes after treatments. DNA fragmentation was analyzed by agarose gel electrophoresis and fluorescence microscopy. RESULTS: The cells deprived of aFGF and serum were exposed to manoalide 1-4 mumol. L-1 for 48 h, detachment and DNA fragmentation of these cells were suppressed. At 7 mumol. L-1, manoalide promoted detachment and DNA fragmentation of VEC. CONCLUSION: manoalide 2 mumol.L-1 inhibited, but 7 mumol.L-1 promoted, apoptosis of VEC.     

Inhibition of serum deprivation-induced PC12 cell apoptosis by tanshinone II A.

AIM: To study the effect of tanshinone II A (Tan II A) on PC12 cell apoptosis induced by serum deprivation. METHODS: PC12 cell survival was measured by MTT assay. The DNA content and percentage of apoptosis were monitored by flow cytometry and DNA fragmentation was analyzed by agarose gel electrophoresis. RESULTS: Serum-free (12 h) medium induced apoptosis in PC12 cells. When the cells had been treated with Tan II A (0.1 and 1 micromol . L-1) for 12 h, the percentage of PC12 cell apoptosis was greatly decreased to 25.71 % and 4.89 % from 96.07 % in serum deprivation alone group, and DNA fragmentation was prevented. Tan II A (0.01 - 10 micromol . L-1) attenuated the cytotoxic effect of sodium cyanide (20 mmol . L-1), glutamate (0.5 mmol . L-1), and sodium nitroprusside (0.5 mmol . L-1). CONCLUSION: Tan II A prevented PC12 cells from apoptosis induced by serum-free medium.     

Effects of novel safrole oxide derivatives, 1-methoxy-3-(3,4-methylenedioxyphenyl)-2-propanol and 1-ethoxy-3-(3,4-methylenedioxyphenyl)-2-propanol,on apoptosis induced by deprivation of survival factors in vascular endothelial cells

Two safrole oxide derivatives, 1-methoxy-3-(3,4-methylenedioxyphenyl)-2-propanol (MOD) and 1-ethoxy-3-(3,4-methylenedioxyphenyl)-2-propanol (EOD), were newly synthesized as promoters of apoptosis in vascular endothelial cells (VECs). The purpose of this study was to investigate the effects of these two safrole oxide derivatives on cell growth and apoptosis induced by deprivation of survival factors (serum and fibroblast growth factors, aFGF and bFGF) in VECs. Morphological changes were observed with light microscopy. Cell growth was determined by using MTT (3-[4, 5-dimethyl thiazol-2-yl]-2, 5-diphenytetrazolium) method. DNA fragmentation was analyzed by agarose gel electrophoresis and fluorescence microscopy. Apoptosis rate and cell cycle distribution were analyzed by flow cytometry (FCM). The cells deprived of FGF and serum were exposed to MOD 10-40 mg l(-1) for 24 h. Cell growth was suppressed (P<.01), while detachment and DNA fragmentation of these cells were promoted (P<.01). When the cells were treated with MOD30 mg l(-1) for 24 h, apoptosis rate was 21.43% (P<.01). The fact that 66.50% of the cells were trapped in S phase of cell cycle indicated that the cell cycle was blocked at S phase. Treated with EOD 10-40 mg l(-1) for 24 h, the cells were observed; the results showed that VEC growth was inhibited and the apoptosis was triggered (P<.01). At 30 mg l(-1) concentration, EOD blocked 55.22% of the cells at S phase. The data suggested that MOD and EOD might promote apoptosis of VEC by blocking the cell cycle at S phase.     

Daidzin protects PC12 cells from serum deprivation-induced apoptosis

This article examines the effect of daidzin on PC12 cell apoptosis induced by serum-free medium. PC12 cell survival was measured by MTT assay. The DNA content and percentage of apoptosis were monitored by flow cytometry and DNA fragmentation was analyzed by agarose gel electrophoresis. The results showed that serum-free (12 h) medium induced apoptosis in PC12 cells. When the cells had been treated with daidzin (0.1, 1 μM) for 12 h, the percentage of PC12 cell apoptosis was significantly decreased to 12.21 and 4.24% from 91.94% in the group with serum deprivation, and DNA fragmentation was prevented. Daidzin (0.01-10 μM) attenuated the cytotoxic effect of sodium cyanide (20 mM), glutamate (0.5 mM) and sodium nitroprusside (0.5 mM) in a manner dependent on concentration. The results suggested that daidzin prevented PC12 cell from serum free-induced apoptosis.     

Daidzin protects PC12 cells from serum deprivation-induced apoptosis

This article examines the effect of daidzin on PC12 cell apoptosis induced by serum-free medium. PC12 cell survival was measured by MTT assay. The DNA content and percentage of apoptosis were monitored by flow cytometry and DNA fragmentation was analyzed by agarose gel electrophoresis. The results showed that serum-free (12 h) medium induced apoptosis in PC12 cells. When the cells had been treated with daidzin (0.1, 1 w M) for 12 h, the percentage of PC12 cell apoptosis was significantly decreased to 12.21 and 4.24% from 91.94% in the group with serum deprivation, and DNA fragmentation was prevented. Daidzin (0.01-10 w M) attenuated the cytotoxic effect of sodium cyanide (20 mM), glutamate (0.5 mM) and sodium nitroprusside (0.5 mM) in a manner dependent on concentration. The results suggested that daidzin prevented PC12 cell from serum free-induced apoptosis.     

Peplomycin induces G1-phase specific apoptosis in liver carcinoma cell line Bel-7402 involving G2-phase arrest

AIM: To investigate the mechanism of peplomycin (PEP)-induced apoptosis in liver carcinoma cell line (Bel-7402).METHODS: Growth inhibition by PEP was analyzed using 3- 4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazoliumbromide (MTT) assay. Apoptotic cells were detected using Hoechest 33258 staining, and confirmed by flowcytometric analysis and DNA fragmentation analysis. The expression of cyclin A and B 1 were determined by flowcytometry and Western blot. Annexin V assay was measured by flow cytometric analysis. RESULTS: PEPinduced apoptosis and then inhibited cell proliferation in liver carcinoma cell line Bel-7402. Cells treated with PEP50 μmol/L for 15 h were arrested in G2-phase with dramatical expression of cyclin A and a little change in cyclin B 1.Almost all the apoptosis occurred in cells undergoing the G1-phase after treatment for 24 h. CONCLUSION:Peplomycin induced G1-phase specific apoptosis in Bel-7402 involving G2-phase arrest.     

Nonylphenol diethoxylate inhibits apoptosis induced in PC12 cells

Nonylphenol and short‐chain nonylphenol ethoxylates such as NP₂EO are present in aquatic environment as wastewater contaminants, and their toxic effects on aquatic species have been reported. Apoptosis has been shown to be induced by serum deprivation or copper treatment. To understand the toxicity of nonylphenol diethoxylate, we investigated the effects of NP₂EO on apoptosis induced by serum deprivation and copper by using PC12 cell system. Nonylphenol diethoxylate itself showed no toxicity and recovered cell viability from apoptosis. In addition, nonylphenol diethoxylate decreased DNA fragmentation caused by apoptosis in PC12 cells. This phenomenon was confirmed after treating apoptotic PC12 cells with nonylphenol diethoxylate, whereas the cytochrome c release into the cytosol decreased as compared to that in apoptotic cells not treated with nonylphenol diethoxylates. Furthermore, Bax contents in apoptotic cells were reduced after exposure to nonylphenol diethoxylate. Thus, nonylphenol diethoxylate has the opposite effect on apoptosis in PC12 cells compared to nonylphenol, which enhances apoptosis induced by serum deprivation. The difference in structure of the two compounds is hypothesized to be responsible for this phenomenon. These results indicated that nonylphenol diethoxylate has capability to affect cell differentiation and development and has potentially harmful effect on organisms because of its unexpected impact on apoptosis. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1389–1398, 2016.     

蓖麻蛋白诱导HeLa细胞凋亡的分子机制(英文)

目的:研究蓖麻蛋白引起的Hela细胞凋亡的形态变化及机制。方法:扫描电镜,透射电镜,Western blot,细胞周期分析、细胞毒性和细胞相对存活率测定。结果:蓖麻蛋白0.05μmol·L~(-1)引起HeLa细胞发生典型的凋亡。凋亡细胞主要表现为胞浆膜起泡,核染色质浓缩,形成新月状核或膜包裹核染色质的凋亡小体;Western blot未检测到p53、Bax和ICE的p20活性亚基,而检测到CPP32的p17活性亚基,CPP32活性升高,而ICE活性无显著改变。结论:CPP32参与了蓖麻蛋白诱导的HeLa细胞凋亡过程。