脂质立方液晶设计制备与表征评价研究进展

目的：研究伊拉地平（ ISR ）对1-甲基-4-苯基吡啶离子（ MPP＋）损伤的PC12细胞的保护作用及可能机制。方法MPP ＋处理PC12细胞建立帕金森病细胞模型;4-甲基偶氮唑蓝（ MTT）比色法检测细胞存活率;双氯荧光黄乙酸乙酯（ DCFH-DA）染色流式细胞术检测细胞内活性氧（ ROS）的生成;JC-1染色流式细胞术检测细胞线粒体膜电位（ MMP）。结果1 mmol · L－1MPP＋处理PC12细胞24 h后能明显抑制细胞生长（P＜0．01）;降低线粒体膜电位;ROS含量增加。2μmol· L－1伊拉地平预处理后, PC12细胞存活率显著增加（ P＜0．01）;线粒体膜电位升高;ROS生成减少。结论伊拉地平对MPP＋损伤的PC12细胞具有保护作用,其作用机制可能与维持线粒体正常膜电位,稳定线粒体功能,阻止线粒体氧化应激发生有关。     

1-甲基-4-苯基吡啶离子诱导大鼠肾上腺嗜铬细胞瘤PC12细胞发生铁死亡的实验观察

目的 进行1-甲基-4-苯基吡啶离子(1-methyl-4-phenyl pyridine,MPP+)诱导大鼠肾上腺嗜铬细胞瘤PC12细胞铁死亡(Ferroptosis)的研究.方法 采用噻唑蓝法(MTT)检测细胞存活率和筛选铁死亡抑制剂(Ferrostatin-1)的最佳作用浓度;倒置显微镜观察Ferrostatin-1对PC12细胞的保护作用;MDC染色检测细胞自噬;ELISA法检测caspase-3的酶活性;流式细胞术检测活性氧ROS.结果 1 mmol·L-1 MPP+对PC12细胞有明显抑制作用;5 μmol·L-1 Ferrostatin-1能够明显提高PC12细胞的存活率;倒置显微镜观察结果表明Ferrostatin-1预保护后,PC12细胞损伤明显减少;MDC染色检测细胞自噬和ELISA法检测caspase-3的酶活性结果显示Ferrostatin-1对MPP+诱导的PC12细胞自噬和凋亡的保护作用不明显;ROS活性氧检测结果显示ROS在胞内增加,可能发生氧化应激,加Ferrostatin-1后ROS荧光强度减弱.结论 Ferroptosis能够显著抑制MPP+诱导的PC12细胞损伤,且Ferrostatin-1对细胞的自噬和凋亡作用不明显,推测出MPP+诱导PC12细胞的损伤可能有Ferroptosis的存在.     

Protective effect of tubuloside B on TNFalpha-induced apoptosis in neuronal cells.

AIM: To investigate the neuroprotective effect of tubuloside B, one of the phenylethanoids isolated from the stems of Cistanche salsa, on tumor necrosis factor-alpha (TNFalpha)-induced apoptosis in SH-SY5Y neuronal cells. METHODS: Cell viability was analyzed using MTT assay. Apoptotic cells were detected using Hoechst33342 staining, and confirmed by DNA fragmentation and flow cytometric analysis. The activity of caspase-3 was measured with special assay kit. The concentration of free intracellular calcium was determined with the probe Indo-1 by spectrometer. The level of intracellular reactive oxygen species and the potential of mitochondrial membrane were determined by laser scanning confocal microscopy (LSCM) combined with fluorescence probe H2DCFDA or JC-1 respectively. RESULTS: SH-SY5Y cells treated with TNFalpha 100 microg/L for 36 h showed typical morphological changes of apoptosis. DNA ladder could be observed by agarose gel electrophoresis. The highest percentage of apoptotic cells accumulated to 37.5 %. Following 36 h treatment with TNFalpha, accumulation of intracellular ROS and [Ca2+]i and decrease in mitochondrial membrane potential were observed, and caspase-3 activity increased by about five-fold compared with controls. However, pretreatment with tubuloside B (1, 10, or 100 mg/L) for 2 h attenuated the TNFalpha-mediated apoptosis. The antiapoptotic action of tubuloside B was partially dependent on an anti-oxidative stress effects, maintain of mitochondria function, decrease of concentration of free intracellular calcium and inhibition of caspase-3 activity. CONCLUSION: Tubuloside B has the neuroprotective capacity to antagonize TNFalpha-induced apoptosis in SH-SY5Y cells and may be useful in treating some neurodegenerative diseases.     

没药甾酮对H2O2损伤PC12细胞的保护作用

探讨没药甾酮(guggulsterone)对氧化应激损伤PC12细胞的保护作用。以过氧化氢(hydrogen peroxide，H₂O₂)损伤PC12细胞为氧化应激损伤模型， 维生素E为对照， 采用四甲基偶氮唑蓝［3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide，MTT］法检测细胞增殖状况； 试剂盒检测乳酸脱氢酶(lactate dehydrogenase,LDH)及一氧化氮(nitric oxide，NO)的释放； DCFH法和Fura 2-AM法检测细胞内活性氧(reactive oxygen species，ROS)和Ca²⁺的含量； 碘化丙啶(propidium iodide，PI)染色流式细胞术(flow cytometry，FCM)检测细胞凋亡； 罗丹明123(rhodamine 123，Rh 123)染色FCM检测细胞线粒体膜电位(mitochondrial membrane protential，MMP)。结果表明， 没药甾酮(0.1～10 μmol·L^-1)可使200 μmol·L^-1 H₂O₂作用24 h后的PC12细胞生长抑制率下降； 细胞外LDH和NO， 细胞内ROS和Ca²⁺含量降低； 明显抑制200 μmol·L^-1 H₂O₂作用12 h后诱导的PC12细胞凋亡和线粒体膜电位降低作用，没药甾酮(0.1～10 μmol·L^-1)使细胞凋亡率由24.3%下降至18.4%、 15.9%、 11.8%。实验结果表明, 没药甾酮对氧化应激损伤PC12细胞具有保护作用， 其机制可能为降低细胞内ROS含量， 进而抑制LDH和NO释放， 降低细胞内Ca²⁺含量， 升高线粒体膜电位,减少细胞凋亡。     

毛蕊花苷对MPP~+诱导的SHSY5Y细胞凋亡的保护作用

目的观察肉苁蓉提取物毛蕊花苷对MPP+诱导的SHSY5Y细胞损伤的影响。方法用MTT法检测细胞存活率,以流式细胞仪检测细胞内活性氧的产生和线粒体膜电位的变化,以及细胞凋亡的发生,并用荧光酶标仪测定caspase-3的活性,蛋白印迹测定Bcl-2的表达水平。结果200μmol·L-1MPP+处理细胞24h降低细胞的存活率;诱导细胞发生凋亡,凋亡率达38.9%;细胞内活性氧水平及caspase-3的活性升高;而线粒体膜电位却明显降低。而预先给予0.1、1或者10mg·L-1浓度的毛蕊花苷处理细胞12h,可提高细胞存活率;流式细胞仪检测凋亡率分别降低到29.5%,15.3%和8.6%,而且细胞内活性氧的水平明显降低,并可逐渐恢复线粒体的高能量状态;caspase-3的活性不断降低,Bcl-2的表达水平增高,并呈现一定的剂量依赖性。结论毛蕊花苷能抑制MPP+诱导的SHSY5Y细胞凋亡,其神经细胞保护作用可能与其降低细胞内活性氧水平,维持线粒体膜电位的高能状态和抑制caspase-3的活性有关。     

Effect of hydrogen sulphide on beta-amyloid-induced damage in PC12 cells

1. Hydrogen sulphide (H(2)S) is a well-known cytotoxic gas. Recently, H(2)S has been shown to protect neurons against oxidative stress caused by glutamate, peroxynitrite and HOCl. Considerably lower H(2)S levels have been reported in the brain of Alzheimer's disease (AD) patients with accumulation of beta-amyloid (A beta). 2. The aim of present study was to explore the cytoprotection by H(2)S against A beta(25-35)-induced apoptosis and the molecular mechanisms underlying this effect in PC12 cells. 3. Our findings indicated that A beta(25-35) significantly reduced cell viability and induced apoptosis of PC12 cells, along with dissipation of the mitochondrial membrane potential (MMP) and overproduction of reactive oxygen species (ROS). 4. Sodium hydrosulphide (NaHS), an H(2)S donor, protected PC12 cells against A beta(25-35)-induced cytotoxicity and apoptosis not only by reducing the loss of MMP, but also by attenuating the increase in intracellular ROS. 5. The results of the present study suggest that the cytoprotection by H(2)S is related to the preservation of MMP and attenuation of A beta(25-35)-induced intracellular ROS generation. These findings could significantly advance therapeutic approaches to the neurodegenerative diseases that are associated with oxidative stress, such as AD.     

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.     

Protocatechuic acid from Alpinia oxyphylla against MPP+-induced neurotoxicity in PC12 cells.

An ethyl acetate extract of Alpinia oxyphylla was found to possess neuroprotective activity against 1-methyl-4-phenylpyridinium ion (MPP(+)) induced apotosis and oxidative stress in cultured PC12 cells. From the extract, a phenolic compound was isolated through bioassay-guided fractionation and identified as protocatechuic acid (PCA) by IR, MS, and (1)H and (13)C NMR spectroscopy. It was the first time which was isolated from the kernels of A. oxyphylla. Exposure of PC12 cells to 1mM MPP(+) may cause significant viability loss and apoptotic cell death. PCA stimulated PC12 cellular proliferation and markedly attenuated MPP(+)-induced apoptotic cell death in a dose-dependent manner. By observing the nuclear morphological changes and flow cytometric analysis, PCA showed its significant effect on protecting PC12 cells against MPP(+)-induced apoptosis. Meanwhile, PCA enhanced the activities of superoxide dismutase (SOD) and catalase (CAT) in PC12 cells. In addition, PCA also dose-dependently reduced the hydrogen peroxide (H(2)O(2))- or sodium nitroprusside (SNP)-induced cell death in PC12 cells. The results suggest that PCA may be one of the primary active components in the kernels of A. oxyphylla and provide a useful therapeutic strategy for the treatment of oxidative stress-induced neurodegenerative disease such as Parkinson's disease.     

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.     

Calycopterin promotes survival and outgrowth of neuron-like PC12 cells by attenuation of oxidative- and ER-stress-induced apoptosis along with inflammatory response

There is mounting evidence implicating the role of oxidative stress induced by reactive oxygen species (ROS) in neurodegenerative disease, including Alzheimer's disease. Herein we investigated the neuroprotective potential of a natural flavonoid, calycopterin, against H(2)O(2)-induced cell death in differentiated PC12 cells. We pretreated PC12 cells with 25, 50, and 100 μM calycopterin followed by the addition of H(2)O(2) as an oxidative stress agent. We measured cell viability by the MTT test and found that 50 μM is the best protective concentration of calycopterin. Moreover, we measured six different parameters of neurite outgrowth. Interestingly, we found that calycopterin not only protects PC12 cells against H(2)O(2)-induced apoptosis but also defends against the destructive effect of oxidative stress on the criteria of neural differentiation. Calycopterin decreased ER stress-associated proteins including calpain and caspase-12, and suppressed ERK, JNK, and p38 MAPK phosphorylation. Moreover, calycopterin inhibited H(2)O(2)-induced nuclear translocation of nuclear factor-κB, a known regulator of a host of genes involved in specific stress and inflammatory responses. This observation was perfectly in agreement with the decrease of COX-2 and TNF-α levels. Calycopterin reduced intracellular ROS levels and increased catalase activity. The protective effect of this compound could represent a promising approach for the treatment of neurodegenerative diseases.     

利福平抑制MPP^＋所致PC12细胞凋亡的研究

目的探讨利福平对MPP＋（1-甲基4-苯基吡啶离子）诱导的分化大鼠嗜铬细胞瘤细胞株（ratphaeochmmocytoma,PC12）细胞活性、细胞形态、调亡率的影响及其影响的机制。方法（1）利用MPP^＋诱导分化PCI2细胞建立帕金森病细胞模型;（2）MTT法检测细胞活性,（3）Tunel原位末端标记法检测细胞形态及半定量细胞凋亡率,（4）流式细胞术检测caspase－3激活率。结果（1）MPP＋作用后,细胞生长活性明显受到抑制,凋亡细胞数量增多,调亡率增加,caspase－3激活率明显增高;（2）而经100、200和300μmol／L各浓度利福平预处理后,利福平各浓度组内细胞活性增高,细胞凋亡程度、凋亡率及caspase－3激活率降低,且与利福平作用浓度存在剂量一效应关系。结论利福平可抑制MPP＋所致PCI2细胞的凋亡,这一作用是通过caspase-3途径起作用的,且存在剂量一效应关系。     

神经妥乐平对H2O2致PC12细胞损伤的保护作用

目的 探索神经妥乐平对H2O2诱导的PC12细胞氧化应激损伤的影响及其潜在机制.方法 用CCK-8法检测细胞存活率,以流式细胞术检测细胞氧化损伤的发生、细胞内活性氧的生成及线粒体膜电位的变化,荧光显微镜观察细胞内活性氧的产生,qRT-PCR测定Caspase-3、Bax和Bcl-2 mRNA的表达.结果 PC12细胞存活率随H2O2浓度的增加而逐渐下降.其中,450μM H2O2处理细胞24 h后细胞存活率、凋亡率、坏死率明显降低;细胞内活性氧水平表达明显升高;线粒体膜电位JC-1红/绿荧光比值下降;Bax和Caspase-3的mRNA表达升高,而Bcl-2的mRNA表达下降,以上指标与对照组相比,差异有统计学意义(P＜0.05).而预先给予0.01UN/ml的NTP处理细胞12h可明显提高细胞存活率,降低细胞凋亡率和坏死率,减少细胞内活性氧生成并提高线粒体膜电位,抑制Bax和Caspase-3的mRNA表达,促进Bcl-2 mRNA的表达,以上指标与H2O2组相比,差异有统计学意义(P＜0.05).结论 NTP能抑制H2O2诱导的PC12细胞损伤,其神经细胞保护作用可能与其降低细胞内活性氧水平、维持线粒体膜电位的高能状态和抑制促凋亡基因表达、促进抗凋亡基因表达有关.     

Protocatechuic acid inhibits apoptosis by mitochondrial dysfunction in rotenone-induced PC12 cells.

Protocatechuic acid (PCA), a phenolic compound isolated from the kernels of Alpinia (A.) oxyphylla, showed the significant neuroprotective effects on hydrogen peroxide (H2O2) or MPP+-induced apoptosis in cultured PC12 cells. However, the mechanism how PCA suppresses rotenone-induced neurotoxicity in cultured PC12 cells remains unclear. In this study, we investigated the protective effects of PCA in PC12 cells exposed to rotenone as an in vitro model of mitochondrial dysfunction and apoptotic cell death. The apoptosis in rotenone-induced PC12 cells was accompanied by the loss of mitochondrial membrane potential, the formation of reactive oxygen species (ROS), the total glutathione depletion, activation of caspase-3 and down-regulation of Bcl-2. In contrast, PCA markedly attenuated the above-mentioned mitochondrial dysfunction in a dose-dependent manner. Taken together, these results suggest that treatment of PC12 cells with PCA can block rotenone-induced apoptosis via ameliorating the mitochondrial dysfunction.     

Protocatechuic acid suppresses MPP+ -induced mitochondrial dysfunction and apoptotic cell death in PC12 cells.

Protocatechuic acid (PCA), a phenolic compound isolated from the kernels of Alpinia (A.) oxyphylla, showed antioxidant neuroprotective effect in our previous study. Here, we investigated the effect of PCA on the MPP(+)-induced mitochondrial dysfunction and apoptotic cell death in PC12 cells. The apoptosis in MPP(+)-induced PC12 cells was associated with loss of mitochondrial membrane potential, the formation of reactive oxygen species (ROS), GSH depletion, activation of caspase-3 and down-regulation of Bcl-2. In contrast, treatment of PC12 cells with PCA significantly prevented the above-mentioned mitochondrial dysfunction. Our data pointed to the potential clinical application/use of PCA to overcome neurodegenerative diseases such as Parkinson's disease.     

人参皂苷Rg1可能通过丝裂素活化蛋白激酶途径抑制细胞凋亡人参皂苷Rg1可能通过丝裂素活化蛋白激酶途径抑制细胞凋亡

目的探讨人参皂苷Rg1对MPP⁺诱导细胞凋亡保护作用的可能信号传导途径。方法用吖啶橙-溴化乙锭染色观察SHSY5Y细胞凋亡率,流式细胞仪检测细胞内活性氧ROS水平,Western Blotting法检测JNK(c-jun NH₂-terminal kinase)激酶活性,免疫细胞化学染色法检测裂解的Caspase-3阳性细胞的表达率。结果经10 μmol·L^-1 Rg1或2.5 mmol·L^-1 N-乙酰半胱氨酸预处理后,MPP⁺诱导的SHSY5Y细胞凋亡受到明显抑制,同时细胞内ROS下降,JNK激酶的活性减弱,裂解的Caspase-3阳性细胞表达率下降。结论Rg1可抑制MPP⁺诱导的SHSY5Y细胞凋亡,其作用机制可能是通过清除ROS、减弱JNK激酶的活性,从而减少Caspase-3的激活     

The role of dopamine transporter in selective toxicity of manganese and rotenone

The dopamine transporter has been shown to be the most relevant target site for the specificity of 1-methyl-4-phenylpyridinium ion (MPP+), a neurotoxin for dopaminergic neurons. In contrast, the mechanisms underlying the selective toxicity of manganese and rotenone, potentially toxic agents implicated in dopaminergic neuronal cell death, remain unknown. The aim of this study was to determine the cellular mechanisms of manganese or rotenone uptake in dopaminergic cells via the dopamine transporter. PC12 cells overexpressing the dopamine transporter, which were exposed to 10microM MPP+, showed extensive DNA fragmentation, a biochemical hallmark of apoptosis, whereas wild-type PC12 cells or vector-transfected PC12 cells, which were exposed to 5mM MPP+, did not show DNA fragmentation. In contrast, manganese and rotenone induced DNA fragmentation at slightly lower concentrations in PC12 cells overexpressing the dopamine transporter compared to control cells. Dopamine transporter inhibitors, such as mazindol, nomifensine, or GBR12909, inhibited MPP+-induced DNA fragmentation but did not affect manganese- and rotenone-induced DNA fragmentation in PC12 cells overexpressing the dopamine transporter. Finally, manganese accumulated to similar levels in PC12 cells overexpressing the dopamine transporter and control PC12 cells following incubation with manganese chloride. These results suggested that the dopamine transporter dose not confer cytotoxicity to manganese and rotenone.     

FCPR16 induces AMPK-dependent autophagy and provides neuroprotection in SH-SY5Y cells and neurons exposed to MPP~+-induced oxidative damage

OBJECTIVE The cause of Parkinson disease (PD) is generally not clear, but it is considered to be related to excessive oxidative damage. Therefore, the identification of therapeutic targets and compounds with antioxidant damage is a reasonable strategy to slow down the progress of PD. FCPR16 is a novel phosphodiesterase4 inhibitor with little emetic potential. Our previous studies showed that FCPR16 was an effective compound for blocking 1-methyl-4-phenylpyridine(MPP+)-induced oxidative damage in SH-SY5Y cells and neurons. However, the detailed mechanisms underlying its protective effect have not been investigated. The level of oxidative stress in neurons is closely related to the balance of mitochondria mass, while autophagy strongly regulates mitochondrial activity in neurons. Our previous study indicated that inhibition of PDE4 or PDE4 knockdown enhanced the activation of autophagy in microglial cells. While whether PDE4 inhibition mediates autophagy in neurons is largely unknown. As described above, autophagy plays a pivotal role in maintaining redox and mitochondrial homeostasis. We were interested in exploring the impact of PDE4 inhibition on autophagy in neurons. METHODS SH-SY5Y cells and neurons was induced with MPP+to mimic PD cell injury in vitro, and MTT assay was used to investigate the viability effects of FCPR16 (50μmol·L-1) with or without different autophagy inhibitors on MPP+-injured SH-SY5Y cells. Detection of apoptosis was performed by PI staining fluorescence. Lysosomes are essential in autophagy, so LYT Red Stain was used to detect lysosomes in SY5Y cells and neurons. Cells were exposed to Cell ROX Deep Red Reagent to detect intracellular reactive oxygen species (ROS). Mitochondrial membrane potential (Δψm)measurement was executed by 5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolyl-carbocyanineiodide (JC-1).To better detect intracellular autophagy, we used the CYTO-ID Autophagy detection kit to detect the autophagic vacuoles and monitor autophagic flux. The expression of autophagy related proteins and other related signal molecules were demonstrated by Western blot. As relevant indicators of oxidative stress, 3-nitrotyrosine (3-NT) and highly toxic peroxide4-hydroxynonenal (4-HNE) were detected with 3-NT and 4-HNE ELISA kits. RESULTS FCPR16 could significantly decrease the expression of p62, an autophagy substrate, at 6 and 12 h, while FCPR16 enhanced the level of LC3-Ⅱ. Similarly, FCPR16 increased the lysosomes fluorescence and CYTO-ID signal in cells and neurons, while it could be blockby 3-methyladenine (3-MA) and hydroxychloroquine sulfate (HCQ). Simultaneously, Treatment of SH-SY5Y cells with FCPR16 prevented MPP+-induced production of ROS and the decline ofΔψm. Importantly, we also found that FCPR16 phosphorylated and thus activated AMPK in SH-SY5Y cells treated with MPP+. In contrast, blockade of the AMPK pathway with compound C blocked the role of FCPR16 in autophagy enhancement. MPP+-induced a significant increase in PI-positive cells, while FCPR16 decreased the ratio of PI positive cells and 3-MA and compound C could block the protective effect. Additionally, FCPR16 reduced MPP+-induced decline of cell viability, and 3-MA and compound C could block the protective effect. CONCLUSION Deficits in autophagy have been proven to participate the pathology of PD and targeting autophagic function has been viewed as a potential therapeutic strategy for the clearance of toxic proteins (such asα-synuclein) and of impaired mitochondria. this is the first time that PDE4 inhibition has been shown to induce autophagic enhancement both in SH-SY5Y cells and in primary cultured neurons. In addition, our findings indicate that inhibition of PDE4 by FCPR16 protects against MPP+-induced oxidative stress and cellular injury in SH-SY5Y cells and neurons through the activation of AMPK-dependent autophagy. Taken together, these results show that PDE4 is a promising target for developing novel drugs against neuronal apoptosis and FCPR16 may be a potential compound for the prevention and treatment of PD.     

Tetrahydroxystilbene glucoside protects human neuroblastoma SH-SY5Y cells against MPP+-induced cytotoxicity

1-methyl-4-phenylpyridinium (MPP+), an inhibitor of mitochondrial complex I, has been widely used as a neurotoxin for inducing a cell model of Parkinson's disease. This study aimed to evaluate the effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG), an active component extracted from Polygonum multiflorum, on MPP+-induced cytotoxicity in human dopaminergic neuroblastoma SH-SY5Y cells. The results from the MTT and lactate dehydrogenase (LDH) assays showed that incubating cells with 500 μM MPP+ for 24 h decreased cell viability and increased LDH leakage, whereas preincubating cells with 3.125 to 50 μM TSG for 24 h protected the cells against MPP+-induced cell damage. Using 2',7'-dichlorofluorescin diacetate (DCFH-DA) and rhodamine 123, respectively, we found that TSG inhibited both the elevation of intracellular reactive oxygen species and the disruption of mitochondrial membrane potential induced by MPP+. In addition, TSG suppressed both the upregulation of the ratio of Bax to Bcl-2 and the activation of caspase-3 induced by MPP+, and TSG inhibited apoptosis as detected by flow cytometric analysis using Annexin-V and propidium (PI) label. These results suggest that TSG may protect neurons against MPP+-induced cell death through improving mitochondrial function, decreasing oxidative stress and inhibiting apoptosis, and this may provide a potentially new strategy for preventing and treating neurodegenerative disorders such as Parkinson's disease.