黄荆子乙酸乙酯提取物诱导结肠癌HT-29细胞凋亡

目的研究黄荆子乙酸乙酯提取物（EVn-50）诱导人结肠癌HT-29细胞凋亡作用。方法用不同浓度的EVn-50处理体外培养的HT-29细胞;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EVn-50以浓度依赖的方式增加HT-29细胞凋亡率（P〈0．05）。10．0tanoVLEVn-50处理HT-29细胞48h导致典型DNA梯形条带形成。10．0umol/LEVn-50以时间依赖方式下调Bcl-2蛋白表达,同时,上调Bax蛋白表达（P〈0．05）。结论EVn-50诱导HT-29细胞凋亡作用与增加Bax／Bcl-2比值有关。     

mir-15a和mir-16-1诱导人淋巴瘤Raji细胞株的凋亡

目的:探讨mir-15a和mir-16-1诱导人淋巴瘤Raji细胞株的凋亡。方法:利用Oligofectamine 2000脂质体法将化学合成的mir-15a和mir-16-1寡核苷酸,随机序列,分别转染Raji细胞。采用间接免疫荧光及流式细胞仪检测Bcl-2蛋白表达情况,半定量RT-PCR检测Bcl-2 mRNA表达水平,台盼蓝拒染法和CCK8法检测mir-15a和mir-16-1对Raji细胞的生长抑制作用,Hoechst染色观察细胞凋亡形态,流式细胞仪—AnnexinV/PI双染色法检测细胞凋亡率。结果:间接免疫荧光法显示Raji细胞经转染mir-15a和mir-16-1寡核苷酸48 h后,Bcl-2蛋白的表达量降低,与空白对照组和随机序列组有显著性差异(P<0.05);RT-PCR法显示各组间Bcl-2 mRNA的表达无显著性差异;台盼蓝拒染法和CCK8法显示,转染后各时间点mir-15a和mir-16-1寡核苷酸组Raji细胞的生长受到抑制,Hoechst染色可见明显凋亡细胞,流式细胞仪—AnnexinV/PI双染色法检测显示,mir-15a组早期凋亡率和晚期凋亡率分别为9.74%和9.65%,mir-16-1组早期凋亡率和晚期凋亡率分别为9.70%和9.34%,明显高于空白对照组和随机对照组。结论:mir-15a和mir-16-1可诱导淋巴瘤细胞凋亡。     

中华补血草多酚诱导白血病细胞HL-60凋亡的研究

采用MTT法研究了中华补血草根多酚提取物对HL-60细胞的抑增殖效果;AO/EB染色、DNA片段化检测和Annexin V-FITC流式细胞术等方法检测了多酚提取物对HL-60细胞诱导凋亡结果.Western blotting检测HL-60细胞株中Bcl-2、Bax蛋白的表达量.结果表明,中华补血草多酚能明显抑制HL-60细胞增殖,诱导HL-60细胞凋亡并具有质量浓度依赖性;Western blotting检测经药物处理的细胞内Bax蛋白显著升高,而Bcl-2蛋白表达水平明显降低,差异具有统计学意义(P<0.05),其凋亡机制可能与下调Bcl-2蛋白、上调Bax蛋白表达量相关.     

垂盆草乙醇提取物诱导HEK 293T细胞凋亡及作用机制

目的:探讨垂盆草乙醇提取物(EESS)对HEK293T细胞凋亡的诱导作用和机制.方法:利用CCK-8法检测垂盆草乙醇提取物对细胞增殖影响,用Hoechst33342染色法及Annexin V-FITC染色流式细胞检测其对细胞凋亡的影响,以Western blot检测其对NF-κB信号通路蛋白及凋亡相关通路蛋白Bcl-2, Bax, Bcl-xL等表达的影响.结果:EESS对细胞增殖具有抑制作用,能促进其凋亡呈剂量依赖性,并显著上调NF-κΒ磷酸化水平及凋亡启动蛋白线粒体细胞色素C(Cyc)蛋白和促凋亡蛋白Bax的表达(P<0.05).结论:垂盆草乙醇提取物能诱导HEK 293T细胞的凋亡,其作用机制与NF-κΒ信号途径及其介导的凋亡启动蛋白Cyt c和促凋亡蛋白Bax的表达上调有关.     

人参水溶性总蛋白对小鼠黑色素瘤细胞B16的增殖抑制的研究及对Bcl-2/Bax表达的影响

摘 要 目的：研究人参水溶性总蛋白对小鼠黑色素瘤细胞的增殖抑制作用及对Bcl-2/Bax表达的的影响。方法：1.MTT法测定人参水溶性总蛋白对黑色素瘤B16细胞株的增殖抑制率。2. RT-PCR（Real-time PCR）法检测Bcl-2、Bax基因的mRNA表达情况。3.Western blot测定凋亡蛋白Bcl-2和Bax的表达。结果：1.人参水溶性总蛋白对小鼠黑色素瘤B16细胞株有明显的增殖抑制作用,其抑制率随着加药浓度的增加而升高,并与加药浓度呈现一定的量效关系。2.随着人参水溶性总蛋白浓度的增高（0,100,500,1000ug/mL）,RT-PCR检测的Bcl-2 mRNA相对表达量逐渐降低,Bax mRNA相对表达量逐渐升高,当加药浓度为1000ug/ml时,Bcl-2的相对表达量最低,为0.20±0.05,Bax的相对表达量最高,为16.83±0.07;与对照组相比,Bcl-2和Bax基因的相对表达量差异均有统计学意义（P＜0.05）。3.经Western blot测定,各组Bcl-2蛋白的表达均低于对照组,各组Bax蛋白表达均高于对照组,且随着加药浓度的增加,Bcl-2蛋白表达量逐渐降低,Bax蛋白表达量逐渐升高。结论：人参水溶性总蛋白可以抑制小鼠黑色素瘤B16细胞株增殖,其分子机制可能与调控Bcl-2/Bax凋亡蛋白的表达有关。     

陡脉冲诱导在体瘤细胞凋亡的实验及分析

检测陡脉冲电场作用下Wistar大鼠在体瘤细胞的Bcl-2和Bax基因编码蛋白表达,探讨陡脉冲诱导在体瘤细胞凋亡的机理.用免疫组织化学法和原位缺口末端标记(TUNEL)法分别检测18例荷有Walker-256瘤株的Wistar大鼠瘤细胞中Bcl-2和Bax蛋白表达阳性率和凋亡细胞百分率.结果表明:1)陡脉冲电场对在体瘤细胞有显著的治疗作用,治疗组可见大量肿瘤坏死组织,对照组瘤细胞呈弥漫性密集分布,浸润性生长;2)TUNEL检测结果:治疗组的细胞凋亡率明显高于对照组;3)Bcl-2、Bax蛋白表达:陡脉冲电场作用后,Bcl-2蛋白表达减弱,Bax蛋白表达增强.陡脉冲电场能诱导瘤细胞凋亡,其分子机制是下调Bcl-2蛋白表达,增强Bax蛋白表达.     

黄连素联合塔斯品碱衍生物对肝癌细胞迁移及凋亡的影响

应用MTT法检测黄连素联合Ta1722对SMMC-7721细胞活力的影响,以金正均法计算q值判断两药相互作用。根据细胞划痕实验与Hoechst染色实验,研究黄连素联合Ta1722对SMMC-7721的迁移及凋亡的影响。使用western blot检测迁移和凋亡相关蛋白表达。黄连素浓度在3.125~25μmol/L范围内、Ta1722浓度在20μmol/L时,两者为协同作用,黄连素联合Ta1722对SMMC-7721细胞的迁移无明显作用,迁移相关蛋白NF-κB,CXCR4,MMP2,MMP9表达无明显变化。黄连素联合Ta1722可诱导SMMC-7721细胞凋亡,上调P53,上调促凋亡蛋白Bax,下调抑凋亡蛋白Bcl-2。黄连素联合Ta1722对SMMC-7721细胞的迁移无明显作用,但可诱导SMMC-7721细胞凋亡,其机制可能是其上调抑癌基因P53的表达,从而导致促凋亡Bax上调,抑凋亡蛋白Bcl-2下调有关。     

五味子酯甲对HepG2细胞增殖和凋亡的影响

目的 观察五味子酯甲(Schisantherin A,SCA)对人肝癌细胞(HepG2)增殖和凋亡的影响.方法 将体外培养的HepG2细胞分为空白对照组(CON)和SCA 25、50、100μmol/L 3个浓度组.给药48 h后,应用MTT法检测细胞存活率;应用Western blot法检测HepG2细胞Bcl-2、Bax和Caspase-3蛋白的表达水平;Hoechst染色观察上述各组细胞凋亡情况.结果 与CON组比较,SCA各组HepG2细胞存活率均显著降低(P<0.01);Bax及Caspase-3蛋白的表达水平显著增加(P<0.01),Bcl-2蛋白及Bcl-2/Bax比值表达显著降低(P<0.05或P<0.01);Hoechst染色显示HepG2细胞出现明显的核浓缩及核碎裂形态.结论 SCA能够抑制HepG2细胞增殖并促进其凋亡,该作用可为开发利用SCA的相关药物提供理论依据.     

表儿茶素没食子酸酯对人鼻咽癌C666-1细胞凋亡的影响

该文观察ECG在人鼻咽癌细胞株C666-1增殖与凋亡中的作用,并初步探讨其机制.人鼻咽癌C666-1细胞给予不同浓度ECG(0~100μmol/L)处理48 h后,采用CCK-8法检测细胞增殖水平,使用TUNEL法和流式细胞仪检测细胞凋亡率,利用Western blotting法检测Bax、Bcl-2、Caspase 3等凋亡相关蛋白的表达变化.结果表明,25μmol/L以上浓度的ECG可抑制C666-1细胞增殖,诱导细胞凋亡;并且ECG可浓度依赖性增加Bax/Bcl-2蛋白比值和剪切的Caspase 3蛋白水平.以上结果表明ECG可抑制人鼻咽癌细胞株C666-1增殖,并诱导细胞凋亡.ECG的作用与其增加Bax/Bcl-2相对蛋白比值进而激活caspase-3有关.     

白藜芦醇类似物对Bel-7404细胞增殖及凋亡的影响

为研制新型抗肿瘤药物,以3,4-二甲氧基苯乙腈和对甲氧基苯甲醛合成白藜芦醇类似物,并以MTT法、流式细胞仪细胞周期分析、Hoechst33258荧光染色及Western blot方法检测合成的白藜芦醇类似物对肝癌Bel-7404细胞的增殖影响及其机制。实验结果证明,所合成的含甲氧基白藜芦醇类似物可通过诱导Bcl-2蛋白表达下调、Bax蛋白表达上调,引起Bel-7404细胞G2/M周期阻滞,并发生细胞凋亡,最终抑制细胞增殖。     

运动对心肌细胞中凋亡调控基因的影响

研究运动对心肌细胞中调控基因Bcl-2、Bax和p53的影响以探讨凋亡调控基因对运动引起心肌细胞凋亡的作用。以大鼠中等运动强度训练、一次性力竭运动和过度训练为运动模型，采用反转录聚合酶链式反应（RT-PCR）技术观察了大鼠心肌细胞中调控基因Bcl-2、Bax和p53mRNA的表达。长期中等强度的运动可造成大鼠心肌细胞中凋亡调控基冈Bcl-2mRNA表达明显增加，可抑制心肌细胞凋亡；而力竭运动和过度训练可引起心肌细胞中Bcl-2mRNA表达下降和调控基冈Bax、p53mRNA表达显著升高以及凋亡调控基因Bcl-2／Bax比值显著下降，可促进心肌细胞凋亡。心肌细胞中凋亡调控基冈Bcl-2、Bax和p53在不同运动后的不同表达对心肌细胞凋亡的发生有明显的调控作用。     

Protective paracrine effect of mesenchymal stem cells on cardiomyocytes

Objective: The aim of this study was to test the protective effect of mesenchymal stem cells (MSCs) on cardiomyocytes in vitro and to investigate the anti-apoptotic signaling pathway. Methods: MSCs from Sprague-Dawley (SD) rats were separated and cultured. MSC medium was collected from MSCs cultured in serum-free Dulbecco’s modified eagle medium (DMEM) under hypoxia. Cultured cardiomyocytes from neonatal SD rats were exposed to hypoxia/reoxygenation (H/R) and treated with MSC medium. The apoptotic cardiomyocytes were stained with Annexin-V-fluorescein isothiocyanate (FITC), Hoechst 33342 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). The mitochondrial transmembrane potential of cardiomyocytes was assessed using a fluorescence microscope. The expression of Bcl-2, Bax, cytochrome C, apoptosis-induced factor (AIF), and caspase-3 was tested by Western blot analysis. Results: Our data demonstrated that MSC medium reduced H/R-induced cardiomyocyte apoptosis, increased the Bcl-2/Bax ratio, and reduced the release of cytochrome C and AIF from mitochondria into the cytosol. Conclusion: MSCs protected the cardiomyocytes from H/R-induced apoptosis through a mitochondrial pathway in a paracrine manner.     

IL-24联合大蒜素体外抗肺癌细胞生长的实验研究

目的探讨抑癌基因IL-24联合大蒜素对肺癌A549细胞凋亡的影响,并初步探讨其作用机制.方法提取前期构建的p DC316-h IL-24-EGFP质粒,转染肺癌A549细胞,分为A组(空白对照组)、B组(单独大蒜素组(40μL/m L))、C组(脂质体+IL-24组)、D组(大蒜素(40μL/m L)+脂质体+IL-24).荧光显微镜下观察质粒的转染情况;流式细胞仪检测细胞凋亡;Western blot法检测p DC316-h IL-24-EGFP、大蒜素和联合用药作用A5 4 9细胞4 8 h后IL-24,Bax,Bcl-2和Caspase-3蛋白表达水平.结果大蒜素(40μL/m L)和p DC316-h IL-24-EGFP单用48 h对A549细胞均有明显抑制作用;大蒜素(40μL/m L)联合p DC3 1 6-h IL-24-EGFP比单用p DC3 1 6-h IL-24-EGFP作用更强(P0.05);大蒜素(40μL/m L)、p DC316-h IL-24-EGFP和两药联用48 h后,A549细胞凋亡率分别为32.7%,39.4%和68.8%;大蒜素(40μL/m L)和/或IL-24作用于A549细胞48 h后,Bax/Bcl-2的比值及Caspase-3表达均上调,联合用药组效果尤其明显.结论p DC316-h IL-24-EGFP联合大蒜素可协同抑制人肺癌A549细胞生长,机制可能是通过调节基因Bax/Bcl-2的表达比率,并上调Caspase-3的表达,进而诱导A549细胞的凋亡.     

Treatment of PC-3 cells with ultrasound combined with microbubbles induces distinct alterations in the expression of Bcl-2 and Bax

The objective of the present study was to investigate whether ultrasound combined with microbubbles induces apoptotic cell death in androgen-independent prostate cancer cells and to identify the probable mechanism. We used ultrasound in continuous wave mode with a frequency of 21 kHz and a spatial-average temporal-average intensity of 46 mW/cm². Ultrasound combined with microbubbles (200 μL SonoVue?) was used to treat androgen-independent human prostate cancer PC-3 cells for 30 s. PC-3 cells were divided into three groups: the control group, the ultrasound group and the ultrasound combined with microbubbles group. Immediately after treatment, trypan blue exclusion was used to assess cell viability. Cell apoptosis at 24 h after treatment was measured using transmission electron microscopy and flow cytometry. Western blotting was used to evaluate the expression of the apoptosis-related proteins, Bcl-2 and Bax. Ultrasound combined with microbubbles had a minimal effect on the viability of PC-3 cells and induced minimal levels of cell lysis. The level of apoptosis in PC-3 cells induced by this modality was significantly higher than in controls (12.77 ± 0.31% vs. 2.56 ± 0.22%, P<0.01). Treatment with ultrasound combined with microbubbles increased the expression of Bax, a pro-apoptotic protein, and decreased the expression of Bcl-2, an anti-apoptotic protein. It was concluded that ultrasound combined with microbubbles induces apoptotic cell death in human prostate cancer PC-3 cells through down-regulation of Bcl-2 and up-regulation of Bax.     

Effect of thrombin on the apoptosis of hippocampal neurons in vitro

Hippocampal neurons were treated by thrombin and thrombin receptor activating peptides (TRAP). Cell survival rate was decreased in a dose-dependent manner by MTT assay. The numbers of apoptotic cell and apoptotic rate of hippocampal neurons treated by different concentrations of thrombin were increased in a dose-dependent manner by terminal deoxynucleotidyl transferase (TdT) mediated dUTP-biotin nick end-labeling (TUNEL) method and Flow Cytometry. When the concentration of thrombin is 40 U/mL, TUNEL positive cells and apoptotic rate of hippocampal neurons reached peak value, were 27.3±4.0 and (29.333±4.633)%, respectively. Immunocytochemistry assay show that Bcl-2 protein expression was down-regulated and Bax protein expression was up-regulated with the concentration of thrombin increased. TRAP can mimic the effect of thrombin to induce apoptosis on hippocampal neurons. These data demonstrated that thrombin induced hippocampal neuron apoptosis in a dose-dependent manner through activating protease-activated protein-1 (PAR-1). The change in expression of Bcl-2 and Bax was related with the effect of high concentration thrombin induced apoptosis on hippocampal neurons. Foundation item: Supported by the Natural Science Foundation of Hainan Province (N30215) Biography: YANG Wen-qiong (1968-), female, Ph.D. candidate, research direction: cerebrovascular disease.     

铅对小鼠睾丸生精细胞凋亡及Caspase-3,Bcl-2和Bax基因表达的影响

为了研究铅致小鼠睾丸生精细胞凋亡及其对Caspase-3,Bax和Bcl -2基因表达的影响.25只雄性小鼠随机分成5组:0h对照组、12h染铅组、24h染铅组、36h染铅组和48h染铅组,每组5只,实验组按照100mg/kg的剂量腹腔注射醋酸铅溶液,5组小鼠分别于0h、12h、24h、36h、48h后脊柱拉断处死,取其睾丸组织,用苏木精-伊红染色及免疫组化技术检测睾丸组织细胞凋亡及凋亡相关蛋白Caspase -3,Bcl -2和Bax的表达.结果显示:细胞凋亡相关基因Caspase -3,Bcl -2和Bax的表达较对照组差异显著,Caspase -3和Bax在染铅毒后表达量一直上升,Bcl -2的表达量一直下降,差异性随染铅时间的不同而有变化.铅负荷至一定程度时可致小鼠睾丸细胞凋亡,且呈一定量效关系(P＜0.01),这种作用可能与Bcl -2基因低表达和Caspase-3与Bax基因高表达有关.     

积雪草苷调控miR-342-5p/AQP3轴保护IL-1β诱导的软骨细胞损伤

目的 探讨积雪草昔(Ass)对白细胞介素1B(IL-1B)诱导的软骨细胞损伤的影响及作用机制。方法 不同 浓度的Ass作用IL-1B诱导软骨细胞24 h,流式细胞仪检测细胞凋亡,酶联免疫吸附法检测细胞培养上清液中白细 胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平,实时荧光定量PCR检测细胞中miR-342-5p和水通道蛋白3 (AQP3)mRNA表达水平,Western Blot法检测B淋巴细胞瘤-2 (Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)和AQP3蛋 白表达水平。双荧光素酶报告基因实验验证miR-342-5p与AQP3靶向关系。转染miR-342-5p抑制剂或AQP3过表达载体至软骨细胞,上述相同方法检测抑制miR-342-5p表达或AQP3过表达对IL-1β诱导的软骨细胞凋亡及IL- 6和TNF-α表达的影响。结果 Ass可抑制IL-1β诱导的软骨细胞凋亡率、Bax蛋白、IL-6和TNF-α表达及miR- 342-5p表达(P <0. 05),促进Bcl-2蛋白、AQP3 mRNA和蛋白表达(P <0.05)。miR-342-5p在软骨细胞中负调控 AQP3表达。抑制miR-342-5p或AQP3过表达后,IL-1&诱导的软骨细胞凋亡率、Bax蛋白、IL-6和TNF-α表达降低 (P <0. 05) ,Bcl-2蛋白表达升高(P <0.05)。抑制AQP3表达逆转了抑制miR-342-5p对IL-1β诱导的软骨细胞凋 亡、Bax和Bcl-2蛋白及IL-6和TNF-α表达的影响。结论 Ass可抑制IL-1β诱导软骨细胞凋亡和炎症反应,其可 能通过调控miR-342-5p/AQP3保护软骨细胞损伤。     

Involvement of Oxidative Stress and Down-Regulation of Bcl-2 in Arachidonic Acid-Induced Apoptosis in HUVECs

0IntroductionAraacicdhi,diosn fiocu ancidd p(reAdAo)m i,naanntelsys eantt itahle p sonl-y2u npsoastiutiroante doff actetl-ylular phospholipids . Normal free AAconcentrationin humanblood ranges from5 .8μmol/Lto 49 .3μmol/L[1].It is re-leased mostlythroughactivation of phospholipase A2by physi-ological and pathological sti muli[2]. Free AAcan be metabo-lizedinto various eicosanoids via specific enzymes such as cy-clooxygenases ( COX) , lipoxygenase and cytochromesP-450[3]. During AA met…