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11.
Yan-Wei Hu Chun-Yu Liu Chong-Min Du Jian Zhang Wen-Qian Wu Zhen-Lun Gu 《Journal of ethnopharmacology》2009
Aim of the study
Flavonoids extracted from the seeds of Astragalus complanatus R.Br. reduce the proliferation of many cancer cells. The present study was carried out to evaluate the effects of these flavonoids from Astragalus complanatus (FAC) on human hepatocarcinoma cell viability and apoptosis and to investigate its mechanisms of action in SMMC-7721 cells.Materials and methods
Cell viability was measured using the MTT assay. To detect apoptotic cells, SMMC-7721 cells treated with FAC were stained with Hoechst 33258 and subjected to agarose gel electrophoresis. Quantitative detection of apoptotic cells was performed by flow cytometry. The effects of FAC on apoptosis and cell cycle regulatory genes and proteins in SMMC-7721 cells were examined using an S series apoptosis and cell cycle gene array and Western blot analysis.Results
The growth of SMMC-7721 and HepG2 cells was inhibited by treatment with FAC. Cell death induced by FAC was characterized by nuclear condensation and DNA fragmentation. Moreover, the cell cycle was arrested in the G0/G1 and S phases in FAC-treated SMMC-7721 cells. A sub-G1 peak with reduced DNA content was also formed. The activity of caspase-3 was significantly increased following FAC treatment. Microarray data indicated that the expression levels of 76 genes were changed in SMMC-7721 cells treated with FAC: 35 genes were up-regulated and 41 were down-regulated. Western blot analysis showed that caspase-3, caspase-8, Bax, P21, and P27 protein levels in SMMC-7721 cells were increased after 48 h of FAC treatment, while cyclinB1, cyclinD1, CDK1, and CDK4 protein levels were decreased.Conclusions
These results suggest that FAC may play an important role in tumor growth suppression by inducing apoptosis in human hepatocarcinoma cells via mitochondria-dependent and death receptor-dependent apoptotic pathways. 相似文献12.
蒋凯|张靖岩|劳华毅|曹永刚|佟立权|张峰 《中国普通外科杂志》2013,22(1):49-53
目的:探讨氨溴索对大鼠肝脏缺血再灌注损伤(I/RI)的保护作用及其机制。方法:18只雄性Wistar大鼠被随机均分为假手术组、肝I/RI模型组(模型组)、氨溴索预处理+肝I/RI模型组(氨溴索预处理组)。肝I/RI模型采用阻断入肝血流30 min后再灌注方法诱导,氨溴索预处理组于缺血前20 min尾静脉注射氨溴索(100 mg/kg),而模型组给予等体积生理盐水。术后6 h处死大鼠,检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平和肝组织病理学改变,同时检测肝组织超氧化物岐化酶(SOD),谷胱甘肽(GSH),丙二醛(MDA)含量,caspase-3的活化水平。结果:与假手术组比较,模型组与氨溴索预处理组血清ALT和AST水平明显升高(均P<0.05);肝组织出现明显的病理学改变;肝组织SOD和GSH含量明显下降,而MDA水平明显升高(均P<0.05);肝组织caspase-3活化水平明显升高(均P<0.05)。与模型组比较,氨溴索预处理组以上各项指标的变化均明显减弱(均P<0.05)。结论:氨溴索预处理能减轻大鼠肝脏I/RI,机制可能与其调控抗氧化和抗凋亡信号通路有关。 相似文献
13.
Caspase-8在5氟尿嘧啶诱导肝癌细胞凋亡中的作用 总被引:4,自引:0,他引:4
目的探讨 5氟尿嘧啶 (5 Fu)诱导肝癌细胞凋亡与caspase 8活性变化的关系。方法采用caspase 8荧光检测试剂盒检测HepG2细胞凋亡过程中caspase 8活性变化。流式细胞仪检测加入caspase 8抑制剂IETD FMK后 5 Fu诱导的HepG2细胞凋亡百分率的变化。结果不同浓度的 5 Fu均可诱导HepG2细胞caspase 8活性升高 ,高浓度与低浓度比较差异有显著意义 (P <0 0 0 1)。肝癌细胞的caspase 8活性随 5 Fu作用时间延长而逐步升高 ,至 16h后达到高峰。IETD FMK能阻断caspase 8活化而抑制 5 Fu诱导的HepG2细胞凋亡。结论 5 Fu通过caspase 8信号传导通路诱导肝癌细胞凋亡 ;5 Fu诱导caspase 8活性升高有浓度与时间依赖性 相似文献
14.
The cathepsin B inhibitor, benzyloxycarbonyl-phenylalanine-alanine-chloromethylketone (z-FA-CMK) was found to be toxic and readily induced cell death in the human T cell line, Jurkat, whereas two other analogs benzyloxycarbonyl-phenylalanine-alanine-fluoromethylketone (z-FA-FMK) and benzyloxycarbonyl-phenylalanine-alanine-diazomethylketone (z-FA-DMK) were not toxic. The toxicity of z-FA-CMK requires not only the CMK group, but also the presence of alanine in the P1 position and the benzyloxycarbonyl group at the N-terminal. Dose–response studies showed that lower concentrations of z-FA-CMK induced apoptosis in Jurkat T cells whereas higher concentrations induced necrosis. In z-FA-CMK-induced apoptosis, both initiator caspases (-8 and -9) and effector caspases (-3, -6 and -7) were processed to their respective subunits in Jurkat T cells. However, only the pro-form of the initiator caspases were reduced in z-FA-CMK-induced necrosis and no respective subunits were apparent. The caspase inihibitor benzyloxycarbonyl-valine-alanine-aspartic acid-(O-methyl)-fluoromehylketone (z-VAD-FMK) inhibits apoptosis and caspase processing in Jurkat T cells treated with low concentration of z-FA-CMK but has no effect on z-FA-CMK-induced necrosis and the loss of initiator caspases. This suggests that the loss of initiator caspases in Jurkat T cells during z-FA-CMK-induced necrosis is not a caspase-dependent process. Taken together, we have demonstrated that z-FA-CMK is toxic to Jurkat T cells and induces apoptosis at low concentrations, while at higher concentrations the cells die of necrosis. 相似文献
15.
Is the Cell Death in Mesial Temporal Sclerosis Apoptotic? 总被引:6,自引:0,他引:6
Uysal H Cevik IU Soylemezoglu F Elibol B Ozdemir YG Evrenkaya T Saygi S Dalkara T 《Epilepsia》2003,44(6):778-784
PURPOSE: Mesial temporal sclerosis (MTS) is characterized by neuronal loss in the hippocampus. Studies on experimental models and patients with intractable epilepsy suggest that apoptosis may be involved in neuronal death induced by recurrent seizures. METHODS: We searched evidence for apoptotic cell death in temporal lobes resected from drug-resistant epilepsy patients with MTS by using the terminal deoxynucleotidyl transferase (TdT) and digoxigenin-11-dUTP (TUNEL) method and immunohistochemistry for Bcl-2, Bax, and caspase-cleaved actin fragment, fractin. The temporal lobe specimens were obtained from 15 patients (six women and nine men; mean age, 29 +/- 8 years). RESULTS: Unlike that in normal adult brain, we observed Bcl-2 immunoreactivity in some of the remaining neurons dispersed throughout the hippocampus proper as well as in most of the reactive astroglia. Bax immunopositivity was increased in almost all neurons. Fractin immunostaining, an indicator of caspase activity, was detected in approximately 10% of these neurons. Despite increased Bax expression and activation of caspases, we could not find evidence for DNA fragmentation by TUNEL staining. We also could not detect typical apoptotic changes in nuclear morphology by Hoechst-33258 or hematoxylin counterstaining. CONCLUSIONS: These data suggest that either apoptosis is not involved in cell loss in MTS, or a very slow rate of cell demise may have precluded detecting TUNEL-positive neurons dying through apoptosis. Increased Bax expression and activation of caspases support the latter possibility. 相似文献
16.
目的 探讨耐药细胞和药敏细胞中caspase - 3mRNA、蛋白的表达及相互关系 ,试图阐明caspase - 3在诱导耐药细胞凋亡中的作用。方法 利用胃癌耐药细胞系SGC790 1/ 0 8,通过RT -PCR研究caspase - 3的mRNA在细胞毒药物处理耐药细胞0h ,2 4h及药敏细胞 0h ,6h时的表达 ,用Western -blot检测其蛋白在耐药细胞中 0h ,6h ,2 4h ,30h的表达和药敏细胞中 0h ,6h ,2 4h的表达 ,DNAladder检测其特征性凋亡 ,MTT法检测细胞的存活率。结果 耐药细胞和药敏细胞在 0h ,2 4h表达的caspase- 3mRNA无显著性差异 ,蛋白表达在耐药细胞中 6h有微量表达 ,30h出现更小片断 ;药敏细胞在 6h时出现小片断 ,2 4h完全裂解。细胞DNA分析表明耐药细胞 30h出现典型的凋亡谱带 ;应用了caspase - 3抑制剂后细胞凋亡延迟。药敏细胞 6h出现典型的凋亡谱带 ,应用抑制剂后凋亡延迟。结论 提高耐药细胞用药量仍可以使耐药细胞凋亡 ,其凋亡的过程与耐药细胞相同 ,均有caspase - 3的激活 ,而抑制了caspase - 3的活性 ,凋亡可被抑制或是延迟 ,其差别在于耐药细胞需要较高的药物诱导浓度。耐药细胞与药敏细胞的凋亡机制的差别可能在于凋亡发生前的诸多因素产生的差异。 相似文献
17.
Variability in apoptotic response to poliovirus infection 总被引:7,自引:0,他引:7
Romanova LI Belov GA Lidsky PV Tolskaya EA Kolesnikova MS Evstafieva AG Vartapetian AB Egger D Bienz K Agol VI 《Virology》2005,331(2):292-306
In several cell types, poliovirus activates the apoptotic program, implementation of which is suppressed by viral antiapoptotic functions. In such cells, productive infection leads to a necrotic cytopathic effect (CPE), while abortive reproduction, associated with inadequate viral antiapoptotic functions, results in apoptosis. Here, we describe two other types of cell response to poliovirus infection. Murine L20B cells expressing human poliovirus receptor responded to the infection by both CPE and apoptosis concurrently. Interruption of productive infection decreased rather than increased the proportion of apoptotic cells. Productive infection was accompanied by the early efflux of cytochrome c from the mitochondria in a proportion of cells and by activation of DEVD-specific caspases. Inactivation of caspase-9 resulted in a marked, but incomplete, prevention of the apoptotic response of these cells to viral infection. Thus, the poliovirus-triggered apoptotic program in L20B cells was not completely suppressed by the viral antiapoptotic functions. In contrast, human rhabdomyosarcoma RD cells did not develop appreciable apoptosis during productive or abortive infection, exhibiting inefficient efflux of cytochrome c from mitochondria and no marked activation of DEVD-specific caspases. The cells were also refractory to several nonviral apoptosis inducers. Nevertheless, typical caspase-dependent signs of apoptosis in a proportion of RD cells were observed after cessation of viral reproduction. Such "late" apoptosis was also observed in productively infected HeLa cells. In addition, a tiny proportion of all studied cells were TUNEL positive even in the presence of a caspase inhibitor. Degradation of DNA in such cells appeared to be a postmortem phenomenon. Biological relevance of variable host responses to viral infection is discussed. 相似文献
18.
A role of inhibitor of apoptosis (IAP) proteins in increased lymphocyte apoptosis in aged humans 总被引:2,自引:0,他引:2
Gupta S 《Mechanisms of ageing and development》2004,125(2):99-101
Lymphocytes from aged humans show increased death-receptor-mediated apoptosis, which is associated with an increased and early activation of caspases. Inhibitor of apoptosis (IAP) proteins inhibit apoptosis by inhibiting activation and activity of caspases. Therefore, we examine the expression of two of the IAPs, the cIAP-2 and XIAP in lymphocytes from young and aged subjects by Western blotting. Lymphocytes from aged expressed significantly less cIAP2 whereas no difference was observed in XIAP expression between young and aged subjects. These data may suggest that decreased cIAP2 may play a role in increased apoptosis in aged humans. Possible mechanisms for the regulation of IAPs in aging are discussed. 相似文献
19.
Jiménez A Jordà EG Verdaguer E Pubill D Sureda FX Canudas AM Escubedo E Camarasa J Camins A Pallàs M 《Toxicology and applied pharmacology》2004,196(2):223-234
The neurotoxic action of the abuse drugs methamphetamine (METH) and 3,4-methylenedioxymethamphetamine (MDMA) on cerebellar granule neurones (CGNs) culture was examined. Treatment for 48 h with METH or MDMA (1-5 mM) induced a higher decrease in viability than 24 h treatment. z.VAD.fmk (100 microM) but not MK-801 nor NBQX recovered control viability values. In both cases, cell death was characterised as apoptotic rather than necrotic by morphology cell observation. Apoptosis measured by flow cytometry indicated an increase in the hypodiploid population after 48 h treatment with METH and MDMA. Apoptosis was reverted by the presence of z.VAD.fmk (100 microM) but not by 10 microM MK-801 or NBQX. Similar results were obtained by analysing nuclear chromatine condensation. These results ruled out excitotoxic participation in amphetamine derivative-induced neurotoxicity in CGNs. Participation of radical oxygen species (ROS) was evaluated using alpha-tocopherol (1-15 microM) and cytometric studies. The co-treatment with 4 mM METH or MDMA for 48 h partially reverted neurotoxic action and apoptotic features, indicating ROS implication in CGNs death by amphetamine derivatives. Alteration of mitochondrial function induced cytochrome C (Cyt C) release after 48-h treatment with METH and MDMA (4 mM). There was also indication of caspase-3-like activation, measured by immunoanalysis and biochemically. Finally, neurodegenerative action caused by amphetamine derivatives may be prevented by using caspase inhibitors. 相似文献
20.
Purpose: α-Fodrin is a neuronal cytoskeletal protein and a known caspase-3 target. We sought to determine whether caspase-3 cleaves α-fodrin in COH rat retinas and whether this process is reduced by adeno-associated virus (AAV)-induced retinal ganglion cell expression of baculovirus inhibitory repeat-containing 4 (BIRC4), a potent caspase-3 inhibitor.Methods: Ocular hypertension was induced unilaterally in five rat eyes by limbal injection of hypertonic saline. In a similar experiment, ocular hypertension was induced in four eyes pre-treated with an intravitreal injection of AAV-BIRC4 to assess α-fodrin cleavage. Western immunoblotting was performed on all retinas.Results: Caspase-3 cleavage of α-fodrin yields a specific 120 kDa protein fragment. COH retina immunoblots indicated significantly more caspase-3 cleavage of α-fodrin than controls (P<0.01, paired t-test). Inhibition of retinal caspase-3 activity with BIRC4 reduced caspase-3-mediated α-fodrin cleavage compared to controls.Conclusion: This confirms our previous finding of caspase-3 cleavage of α-fodrin in COH retinas and parallels pathology seen in Alzheimer’s disease, in which neurons undergo chronic caspase activation, slow build-up of cleavage products, and delayed apoptosis. If caspase activation in glaucoma leads to protracted rather than rapid retinal ganglion cell apoptosis, a much longer therapeutic window exists for apoptosis inhibition with caspase inhibitors such as BIRC4. 相似文献