Stevioside enhances apoptosis induced by serum deprivation in PC12 cells

In the laboratory, using a PC12 cell system, studies have been conducted on the effects of various chemicals on apoptosis, as it is considered to be an essential part of normal development, maintenance, and defense in organisms. Stevioside is a natural sweetener extracted from the leaves of Stevia rebaudiana. Since it is widely used as a sugar replacement, it was decided to evaluate the toxicological effects of low concentrations of stevioside on apoptosis induced by serum deprivation using the PC12 cell system. It was found that based on data from DNA electrophoresis and TUNEL signal assays stevioside enhanced apoptosis induced by serum deprivation. This enhancement was caused by increased expression of Bax and of cytochrome c released into the cytosol. These findings suggest that stevioside affects the regulation of the normal apoptotic condition. Further investigation will be needed to clarify the detailed mechanism of the enhancement due to the treatment with stevioside.     

Neuroprotective effects of chlorogenic acid against apoptosis of PC12 cells induced by methylmercury

Chlorogenic acid (CGA) widely exists in edible and medicinal plants. We aimed to evaluate the effect of CGA on the protection from apoptosis by methylmercury (MeHg) in PC12 cells. Cell viability was evaluated by MTT assay. Apoptosis was assayed by flow cytometry detection. Caspase-3 activity was measured by confocal microscopy. Intracellular GSH levels were determined by bicinchoninic acid protein assay. Intracellular reactive oxygen species (ROS) was assessed by means of chloromethyl-dihydrodichlorofluorescein diacetate. Glutathione peroxidase (GPx) activity was determined by UV. In order to elucidate the action of CGA, the protective effects of CGA were compared to Vit.E. CGA was effective at protecting PC12 cells against MeHg-induced damage in dose-dependent manner. CGA not only suppressed the generation of ROS, the decrease of activity in GPx and the decrease of GSH, but also attenuated caspase-3 activation in PC12 cells by MeHg. CGA eventually protected PC12 cells against MeHg-induced apoptosis. The results highlighted that CGA may exert neuroprotective effects through its antioxidant actions.     

Flupirtine and retigabine prevent -glutamate toxicity in rat pheochromocytoma PC 12 cells

Flupirtine is an analgesic drug thought to have NMDA receptor antagonistic and antiapoptotic effects. We investigated the effects of Ethyl-2-amino-6-(4-(4-fluorbenzyl)amino)-pyridine-3-carbamamic acid, maleate (flupirtine) and the related compound N-(2-amino-4-(4-fluorobenzylamino)-phenyl)-carbamic acid, ethyl ester) (retigabine) (Desaza-flupirtine) on the toxicity of -glutamate and -3,4-dihydroxyphenylalanine ( -DOPA) in rat pheochromocytoma PC 12 cells in vitro. Both drugs (10 μM) markedly decreased nonreceptor-mediated necrotic cell death in PC 12 cultures treated with -glutamate (10 mM) for 72 h. In contrast, apoptosis induced by -DOPA (250 μM) after 48 h was not affected by either substance. While -DOPA elicited massive generation of reactive oxygen intermediates, -glutamate-induced cell death was accompanied by only slightly increased levels of reactive oxygen intermediates. Flupirtine and retigabine exerted anti-oxidative effects in PC 12 cultures independent of their ability to prevent cell death. Further examination of the protective action of flupirtine and retigabine against -glutamate toxicity showed that it had no influence on monoamine oxidase (monoamine: oxygen oxidoreductase (deaminating), EC 1.4.3.4., MAO) activity. Thus, flupirtine and retigabine provided protection against cystine deprivation and -glutamate toxicity but did not protect against -glutamate under cystine-free conditions indicating that both compounds are sufficiently effective to compensate the oxidative stress elicited by cystine deprivation but not excessive activity of monoamine oxidase after -glutamate treatment.     

Deoxynivalenol induces apoptosis in PC12 cells via the mitochondrial pathway

Deoxynivalenol (DON) has broad toxicity in animals and humans. In this study the impact of DON treatment on apoptotic pathways in PC12 cells was determined. The effects of DON were evaluated on (i) typical indicators of apoptosis, including cellular morphology, cell activity, lactate dehydrogenase (LDH) release, and apoptosis ratio in PC12 cells, and on (ii) the expression of key genes and proteins related to apoptosis, including Bcl-2, Bax, Bid, cytochrome C (Cyt C), apoptosis inducing factor (AIF), cleaved-Caspase9, and cleaved-Caspase3. DON treatment inhibited proliferation of PC12 cells, induced significant morphological changes and apoptosis, promoted the release of Cyt C and AIF from the mitochondria, and increased the activities of cleaved-Caspase9 and cleaved-Caspase3. Bcl-2 expression decreased with increasing DON concentrations, in contrast to Bax and Bid, which were increased with increasing DON concentration. These data demonstrate that DON induces apoptosis in PC12 cells through the mitochondrial apoptosis pathway.     

Mechanism of apoptosis induced by copper in PC12 cells

Copper, an essential trace element, induces apoptosis in mammalian cells. However, the precise mechanism of copper-induced apoptosis is still unclear. In this study, to determine the apoptotic pathway initiated by copper treatment, apoptotic factors such as Bax, Bad and Bcl-2, and the caspase family in PC12 cells treated with copper were measured by Western blot and RT-PCR analyses. The expression of Bax, Bad, cytochrome c and caspases 3 and 9 were increased by copper treatment. From these results, two pathways for copper-induced apoptosis were suggested. At first, an increase of Bax induces the release of cytochrome c from the mitochondria into the cytoplasm owing to binding to apoptotic activating caspase 9 leading to the activation of caspases 3. In the other pathway an increase of Bax and reactive oxygen species activates the release of AIF from the mitochondria. The AIF induces apoptosis via a caspase-independent pathway.     

超氧化物歧化酶对浓缩铀诱发PC12细胞凋亡的保护作用

目的 研究超氧化物歧化酶（SOD）对浓缩铀诱导PC12细胞凋亡的保护作用。方法 在PC12细胞中加入浓缩铀DMEM／F12工作液，计算PC12细胞的内照射吸收剂量。结果 SOD可抑制浓缩铀诱发细胞迅速下降及DNA链断裂，同时提高细胞内游离精脒含量。结论 研究表明外源性SOD在清除自由基、抑制凋亡、保护细胞抗浓缩铀诱发损伤中有着重要作用。     

氯化钴诱导PC12细胞凋亡的分子机制

目的确定氯化钴(CoCl2)诱导PC12细胞凋亡的分子机制。方法500μmolLCoCl2诱导PC12细胞24h后,检测活性氧(ROS)生成量的变化以及抗氧化剂N乙酰半胱氨酸(NAC)、二硫代苏糖醇(DTT)对细胞存活率和ROS生成量的影响;琼脂糖凝胶电泳检测NAC、DTT对CoCl2诱导PC12细胞DNA片段化的影响;利用RTPCR法检测凋亡相关基因bclxl和bax在PC12细胞调亡过程中的表达及NAC、DTT对基因表达的影响;化学发光法检测NAC、DTT对Caspase3表达的作用。结果在CoCl2诱导PC12细胞凋亡中,ROS生成量明显上升,为正常时的2.92倍;NAC、DTT可提高细胞存活率,由53.1%上升为94.8%和92.3%(P<0.01),并有效抑制ROS的生成,为正常时的24.2%和82.1%(P<0.01);同时抑制DNA片段化的发生。bclxl在细胞凋亡过程中表达明显下降,bax表达无明显变化;NAC和DTT可使bclxl的表达明显上升。Caspase3在细胞凋亡中被激活,NAC、DTT可抑制Caspase3的表达(P<0.01)。结论ROS介导了CoCl2诱导的PC12细胞凋亡,这一过程与抑制bclxl表达,激活Caspase3有关。     

Oxidative mechanisms contribute to nanosize silican dioxide-induced developmental neurotoxicity in PC12 cells

Neurotoxicity was investigated in nano-SiO₂-treated cultured PC12 cells, an in vitro neuronal cell model, in order to define a relatively safe dose range for its application. The following were observed in the present study: (1) A dose-dependent increase in the level of reactive oxygen species (ROS) with a corresponding decrease in the level of glutathione (R² = 0.965) suggesting 20- and 50-nm SiO₂-induced free radical generation and glutathione depletion. (2) A dose- and time-dependent decrease in cell viability that was associated with elevation of ROS level, especially after 24-h nano-SiO₂ exposure (R² = 0.965), suggesting the role of oxidative stress on nano-SiO₂ induced cell death. (3) An increase in the level of thiobarbituric-acid reactive species that correlated reversely with cell viability of the PC12 cells treated with nano-SiO₂ (R² = 0.945) suggesting nano-SiO₂-induced membrane damage caused by lipid peroxidation. (4) A dose-dependent increase in sub-G1 population in SiO₂-exposed cells along with cell shrinkage and nuclear condensation from morphological examination suggesting nano-SiO₂-induced cell apoptosis. Furthermore, nano-SiO₂ exposure diminished the ability of neurite extension in response to nerve growth factor in treated PC12 cells. In summary, SiO₂ nanoparticle exposure resulted in dose-dependent neurotoxicity in cultured PC12 cells that was probably associated with oxidative stress and induced apoptosis.     

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.     

Aspartame-induced apoptosis in PC12 cells

Aspartame is an artificial sweetner added to many low-calorie foods. The safety of aspartame remains controversial even though there are many studies on its risks. In this study, to understand the physiological effects of trace amounts of artificial sweetners on cells, the effects of aspartame on apoptosis were investigated using a PC12 cell system. In addition, the mechanism of apoptosis induced by aspartame in PC12 cells and effects on apoptotic factors such as cytochrome c, apoptosis-inducing factor, and caspase family proteins were studied by Western blotting and RT-PCR.Aspartame-induced apoptosis in PC12 cells in a dose-dependent manner. In addition, aspartame exposure increased the expressions of caspases 8 and 9, and cytochrome c. These results indicate that aspartame induces apoptosis mainly via mitochondrial pathway involved in apoptosis due to oxigen toxicity.     

丙泊酚对布比卡因诱导的PC12细胞毒性和硫氧还蛋白系统的影响

目的：探讨丙泊酚对布比卡因诱导的PC12细胞毒性的保护作用及内源性硫氧还蛋白（Trx）系统在其中的作用。方法：培养的PC12细胞分成四组，正常对照组、丙泊酚组、布比卡因组、丙泊酚＋布比卡因（PB）组，每组6孔。培养6h和24h后，用MTT比色微量分析细胞存活率，测定上清液乳酸脱氢酶（LDH）活性和细胞内硫氧还蛋白还原酶（TrxR）、活性氧（ROS）活性，RT-PCR检测Trx-1 mRNA和TrxR-1 mRNA表达。结果：与正常PC12细胞相比，布比卡因可显著降低细胞存活率（P〈0．01）和细胞内TrxR活性（P〈0．05），增加上清液中LDH活性和细胞内ROS活性（P〈0．05，P〈0．01），明显降低Trx mRAN和Trx mRAN表达（P〈0．05）；丙泊酚对正常PC12细胞无明显影响；与布比卡因组相比，PB组细胞存活率（P〈0．01）和细胞内Trx活性（P〈0．05）明显增加，上清液中LDH活性和细胞内ROS活性显著降低（P〈0．05，P〈0．01），Trx mRAN和Trx mRAN表达明显增加（P〈0．05）。结论：布比卡因对PC12细胞具有毒性作用可能与降低细胞内TrxR活性、增加ROS活性有关，丙泊酚通过保护细胞内Trx系统的活性及清除ROS来减轻布比卡因诱导的PC12细胞毒性。     

川芎嗪对H_2O_2诱导PC12细胞凋亡的保护作用研究

目的探讨盐酸川芎嗪对H2O2诱导PC12细胞损伤的保护作用。方法采用川芎嗪预处理PC12细胞后,制备H2O2细胞损伤模型,通过四甲基偶氮唑盐(MTT)法检测细胞存活率,测定细胞培养上清液中乳酸脱氢酶(LDH)的活性及细胞活性氧(ROS)水平来反映细胞损伤程度,流式细胞术和蛋白印迹法分析细胞凋亡发生机制。结果川芎嗪预处理后,PC12细胞的存活数提高,LDH释放及ROS水平显著降低(P<0.05),总凋亡率降低,caspase-3活性降低,Bax/Bcl-2比值降低。结论川芎嗪预处理后可以降低H2O2诱导的PC12细胞凋亡。     

海风藤对氯化钴诱导PC12细胞凋亡的保护作用

目的研究海风藤对氯化钴(CoCl2)诱导的PC12细胞凋亡的分子机制,从而确定海风藤对缺氧诱导的神经细胞凋亡的保护作用。方法设立海风藤保护组,CoCl2诱导组和正常细胞对照组;倒置相差显微镜观察细胞形态变化,四甲基偶氮噻唑蓝法(MTT)检测细胞存活率,分光光度法检测培养基中乳酸脱氢酶(LDH)含量和活性氧(ROS)生成量水平变化;化学发光法检测细胞三磷酸腺苷(ATP)生成量和Caspase-3表达量的变化;RT-PCR法检测凋亡相关基因bcl-xl的表达。结果海风藤预保护的PC12细胞在CoCl2诱导后,与CoCl2诱导组相比,细胞形态有明显改善,细胞存活率显著上升,由28.7%变为65.1%(P<0.01);培养基中LDH含量和ROS生成量明显下降,相应吸光度分别由29.8和18.3变为18.33及7.8(P<0.01);细胞ATP释放量明显增加,保护组为诱导组的1.2倍(P<0.01);基因bcl-xl表达上升;Caspase-3在细胞凋亡中的活性被抑制,保护组为诱导组的81%(P<0.01)。结论海风藤能通过抑制氧化应激损伤对线粒体功能破坏,增强bcl-xl的表达,抑制Caspase-3激活等机制提高细胞存活率,达到抑制CoCl2诱导PC12细胞凋亡作用。     

盐酸埃他卡林对谷氨酸所致PC12细胞损伤的保护作用

目的 研究盐酸埃他卡林(Ipt)对谷氨酸(Gli)所致细胞损伤的保护作用。方法 采用培养PC12细胞,MTT法测细胞存活率。结果 Ipt 1×10~(-10)～1×10~(-4)mol·L~(-1)可拮抗Glu介导的神经毒作用,提高细胞存活率,该保护作用可被10 μmol·L~(-1)格列苯脲(ATP敏感性钾通道拮抗剂)所取消。结论 Ipt可拮抗Glu介导的神经毒作用,该保护作用可能与ATP敏感性钾通道相关。     

丹皮酚对过氧化氢诱导的PC12细胞凋亡的抑制作用(英文)

目的探讨丹皮酚对过氧化氢(H2O2)诱导的PC12细胞凋亡的抑制作用及其机制。方法建立H2O2致PC12细胞损伤模型,采用MTT法测定细胞存活率,流式细胞术测定细胞凋亡率及细胞内活性氧含量,化学比色法测定乳酸脱氢酶(LDH)释放量及细胞内丙二醛(MDA)含量。结果PC12细胞经H2O2100μmol.L-1处理10 h可致细胞存活率下降,并能诱导细胞凋亡,LDH释放量及细胞内活性氧和MDA含量明显增加;丹皮酚(12,25和50μmol.L-1)预处理1 h可提高细胞存活率,减少细胞凋亡、LDH释放量及细胞内活性氧和MDA含量。结论丹皮酚对H2O2诱导的PC12细胞凋亡具有抑制作用,该作用可能与其抗氧化作用有关。     

一条新的可诱导PC12细胞转化为交感神经原表型的小分子肽链

目的根据神经生长因子 (NGF)的氨基酸序列及其晶体构象资料 ,对 β NGF进行限制性酶解 ,从而获得关键的功能区域片段。方法选择溴化氰在 9位Met处 ,Trypsin在Arg或Lys处裂解 β NGF ,用SephadexG 5 0色谱、DE5 2离子交换色谱和反相高效液相色谱进行分离纯化。所得片段用PC12细胞测定活性 ,对活性片段进行氨基酸组成及序列分析。结果从 β NGF裂解片段中获得一可诱导PC12细胞分化的活性片段 ,氨基酸组成及序列分析表明 ,此片段由 16肽GEFSVCDSVSVWVGDK与 14肽HWNSYCTTTHTFVK通过一对二硫键连接而成 ,与 β NGF肽链的10～ 2 5和 75～ 88氨基酸残基序列相对应。生物活性分析表明其最佳作用浓度为 0 .0 0 1～ 0 .1ng/ml。结论此实验获得了 β NGF关键的功能区域片段 ,虽然其空间结构和功能的关系尚需研究探讨 ,但它的成功分离和鉴定为合成或表达高活性小分子神经营养物质奠定了重要的基础     

Scutellarin protects PC12 cells from oxidative stress-induced apoptosis

The present study investigated the effects of scutellarin on oxidative stress-induced cell apoptosis in PC12 cells. Exposure of cells to hydrogen peroxide (H₂O₂) triggered a typical apoptosis, as evidenced by DNA fragmentation, DNA loss and externalization of phosphatidylserine (PS). This treatment also caused significant elevation of oxidative stress characterized by intracellular accumulations of reactive oxygen species (ROS) and malondialdehyde (MDA), a product of lipid peroxidation. Preincubation of cells with scutellarin significantly inhibited the fragmentation and loss of DNA, the externalization of PS, and decreased the percentage of cell apoptosis. Also, intracellular accumulations of ROS and MDA resulting from H₂O₂ exposure were significantly reduced by scutellarin. These findings suggest that scutellarin exerts significant protection against oxidative stress-induced apoptosis, which might be beneficial for the prevention and treatment of oxidative stress-mediated disorders.     

Scutellarin protects PC12 cells from oxidative stress-induced apoptosis

The present study investigated the effects of scutellarin on oxidative stress-induced cell apoptosis in PC12 cells. Exposure of cells to hydrogen peroxide (H₂O₂) triggered a typical apoptosis, as evidenced by DNA fragmentation, DNA loss and externalization of phosphatidylserine (PS). This treatment also caused significant elevation of oxidative stress characterized by intracellular accumulations of reactive oxygen species (ROS) and malondialdehyde (MDA), a product of lipid peroxidation. Preincubation of cells with scutellarin significantly inhibited the fragmentation and loss of DNA, the externalization of PS, and decreased the percentage of cell apoptosis. Also, intracellular accumulations of ROS and MDA resulting from H₂O₂ exposure were significantly reduced by scutellarin. These findings suggest that scutellarin exerts significant protection against oxidative stress-induced apoptosis, which might be beneficial for the prevention and treatment of oxidative stress-mediated disorders.     

无肋马尾藻多肽的制备及其促进大鼠肾上腺嗜铬细胞瘤细胞(PC12)分化特性的研究

目的 从无肋马尾藻中提取分离一种生物活性多肽(SFPP),分析其理化特性及对大鼠肾上腺嗜铬细胞瘤PC12细胞分化的促进作用.方法 采用葡聚糖凝胶层析分离和透析脱盐纯化无肋马尾藻的水浸提物,应用Tricine-SDS-PAGE电泳和差示扫描量热法(DSC)分析SFPP的分子量和热变性温度,检测其对PC12细胞分化的促进作...     

Effects of tributyltin chloride on L-DOPA-induced cytotoxicity in PC12 cells

Tributyltin chloride (TBTC) at concentrations of 0.5-1.0 microM inhibits dopamine biosynthesis in PC12 cells. In this study, the effects of TBTC on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced cytotoxicity in PC12 cells were investigated. TBTC at concentrations up to 1.0 microM neither affected cell viability, nor induced apoptosis after 24 or 48 h in PC12 cells. However, TBTC at concentrations higher than 2.0 microM caused cytotoxicity through an apoptotic process. In addition, exposure of PC12 cells to non-cytotoxic (0.5 and 1.0 microM) or cytotoxic (2.0 microM) concentrations of TBTC in combination with L-DOPA (20, 50 and 100 microM) resulted in a significant increase in cell loss and the percentage of apoptotic cells after 24 or 48 h compared with TBTC or L-DOPA alone. The enhancing effects of TBTC on L-DOPA-induced cytotoxicity were concentration- and treatment time-dependent. These data demonstrate that TBTC enhances L-DOPA-induced cytotoxicity in PC 12 cells.