山茱萸羟甲基糠醛对叠氮钠致神经细胞损伤的影响及其作用机制

目的观察从山茱萸中提取的山茱萸羟甲基糠醛(HMF)对线粒体呼吸链复合体IV抑制剂叠氮钠致神经细胞损伤的保护作用,并探讨其作用机制。方法不同剂量的HMF分别与人神经母细胞瘤细胞系SH-SY5Y细胞预孵育24h,洗去培养液,加入50mmolL-1叠氮钠作用4h,用MTT法和乳酸脱氢酶(LDH)漏出率测定细胞存活率,激光共聚焦显微镜观察线粒体膜电位和细胞骨架(微管、微丝)结构,免疫细胞化学法检测β-淀粉样肽(Aβ)的表达。结果12.5～200mg·mL-1的HMF与SH-SY5Y细胞共孵育能明显拮抗叠氮钠的损伤作用,使细胞存活率增高,线粒体跨膜电位升高,并使损伤的微管、微丝结构基本恢复正常,减少Aβ含量。结论HMF能明显拮抗线粒体呼吸链复合体IV缺陷所致的神经细胞损伤,其机制与保护细胞骨架和降低Aβ含量有关,对于防治阿尔采末病等神经退行性疾病有一定应用前景。     

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。     

西红花苷对缺氧复氧损伤的SH-SY5Y细胞线粒体动力学的影响

目的探讨西红花苷对缺氧缺糖(oxygen-glucose deprivation,OGD)损伤的人神经母细胞瘤细胞(SH-SY5Y)线粒体的保护作用及可能的机制。方法 SH-SY5Y细胞OGD损伤同时给予药物,4 h后复氧2 h,测定线粒体膜电位、胞内钙离子浓度;免疫荧光、免疫印迹法检测细胞线粒体Drp1、Opa1、Mfn1、Mfn2含量变化。结果 SH-SY5Y细胞缺氧后线粒体膜电位明显降低,胞内钙离子浓度升高;西红花苷明显提高OGD损伤引起的SH-SY5Y细胞线粒体膜电位,降低胞内钙离子浓度,抑制Drp1表达量,上调Opa1表达量。结论西红花苷对OGD损伤的SH-SY5Y细胞具有明显的保护作用,恢复其线粒体功能,起保护作用机制可能与抑制线粒体分裂融合异常有关。     

钾通道阻断剂部分抑制三氧化二砷诱导的HeLa细胞死亡

目的研究钾通道阻滞剂对三氧化二砷诱导的HeLa细胞死亡的作用。方法采用MTT法评价HeLa细胞的存活情况，采用膜片钳技术记录HeLa细胞的电压依赖性钾电流。结果As₂O₃(5 μmol·L^-1)孵育24 h引起显著的HeLa细胞死亡，As₂O₃(5 μmol·L^-1)孵育24 h后存活的细胞表现明显的电压依赖性钾电流密度增加。+80 mV电压下，As₂O₃(5 μmol·L^-1)孵育组电流密度(61±18) pA/10 pF(n=8)明显高于对照组(38±10) pA/10 pF(n=8,P<005)。As₂O₃诱导的HeLa细胞死亡可被共同孵育钾通道阻滞剂四氨基吡啶(3 mmol·L^-1)或四乙基铵(5 mmol·L^-1)所部分抑制。3 mmol·L^-1四氨基吡啶或5 mmol·L^-1四乙基铵对HeLa细胞无明显细胞毒作用。结论As₂O₃长期处理增加HeLa细胞的电压依赖性钾电流。As₂O₃诱导的HeLa细胞死亡可被钾通道阻滞剂四氨基吡啶或四乙基铵部分抑制。     

五味子酚对氧自由基引起大鼠脑突触体和线粒体损伤的保护作用

以Fe²⁺-半胱氨酸(Cys)为氧自由基生成系统,在体外模仿脑出血或脑外伤引起的氧自由基损伤的模型,观察五味子酚是否对Fe²⁺-Cys引起的大鼠脑突触体和线粒体损伤有保护作用,以探讨Sal用于延缓衰老、防治某些神经系统疾病的可能性。结果显示,与Fe²⁺-Cys共温孵可使脑突触体和线粒体MDA生成量显著增加,线粒体ATPase活性下降。而预先加入Sal(10^{-6mol·L-1)可抑制MDA生成,防止线粒体ATPase活性降低。Sal对Fe2+-Cys引起的线粒体肿胀和膜流动性降低也有明显的保护作用,并能防止Fe2+-Cys所致线粒体和突触体形态的病理性损伤。结果提示,Sal对氧自由基引起的大鼠脑突触体和线粒体损伤有明显保护作用。}     

黄芩苷通过硫氧还原蛋白介导对SH-SY5Y细胞保护作用

目的 研究黄芩苷对过氧化氢(H₂O₂)损伤后的成人神经细胞瘤细胞母细胞SH-SY5Y细胞的影响,进一步探讨黄芩苷保护SH-SY5Y细胞的作用机制. 方法 不同浓度黄芩苷预先处理正常培养的SH-SY5Y细胞24 h,200 μmol.L^-1 H₂O₂损伤上述SH-SY5Y细胞24 h,采用实时荧光定量聚合酶链反应(PCR)技术检测各实验组细胞的硫氧还原蛋白(Trx)mRNA的表达;采用酶联免疫吸附测定(ELISA)技术检测各实验组细胞的Trx 蛋白的表达. 结果 黄芩苷在50～300 μmol.L^-1浓度范围内对SH-SY5Y细胞无细胞毒作用,0,50,100和200 μmol.L^-1黄芩苷组SH-SY5Y细胞存活率分别为(0.410±0.096),(1.304±0.101),(0.899±0.089)和(0.609±0.023),50,100和200 μmol.L^-1均明显高于H₂O₂ 损伤组(0.339±0.073)的存活率,正常组,H2O2损伤组,50 ,100和200 μmol.L^-1黄芩苷组Trx mRNA的表达水平分别为(1.023±0.014),(0.021±0.004),(0.054±0.005),(0.071±0.013)和(0.042±0.004);正常组Trx蛋白表达水平为(26.230±0.857),H₂O₂损伤组为(19.230±0.982),50,100和200 μmol.L^-1黄芩苷组Trx蛋白表达水平分别为(22.440±0.888),(23.020±1.070),(22.330±1.067). 结论 黄芩苷对H₂O₂损伤后的SH-SY5Y细胞有保护作用,其作用机制可能与黄芩苷抑制H2O2诱导的Trx表达下调作用有关,起到抗氧化应激及抗细胞凋亡的作用.     

山茱萸拮抗线粒体缺陷致神经细胞微管及微管相关蛋白表达异常

目的观察中药山茱萸水提液对线粒体呼吸链复合体IV(即细胞色素C氧化酶)抑制剂叠氮钠(NaN3)致神经细胞骨架系统损伤的拮抗作用,探讨山茱萸在防治阿尔采末病中的机制。方法山茱萸水提液(0.125～8g生药·L-1)分别与SH-SY5Y人神经母细胞瘤细胞及原代培养新生大鼠海马神经元共孵育24～48h,加入64mmol·L-1叠氮钠作用4～6h,MTT、LDH法测定细胞存活率,激光共聚焦显微镜观察细胞微管结构及微管相关蛋白表达的变化。结果1～4g·L-1山茱萸能明显拮抗叠氮钠致神经细胞存活率下降;使损伤的微管结构基本恢复正常,微管相关蛋白表达增加,尤以突起内最为明显。结论中药山茱萸能明显拮抗线粒体缺陷致神经细胞损伤,其机制与保护细胞骨架系统有关,对于防治阿尔采末病等神经退行性疾病有一定应用前景。     

羟乙基葛根素对过氧化氢致牛脑微血管内皮细胞损伤的保护作用

目的研究牛脑微血管内皮细胞(BCMEC)过氧化损伤机制并探讨羟乙基葛根素对牛脑微血管内皮细胞损伤的保护作用。方法用MTT法和LDH活性检测测定BCMEC的损伤；倒置相差显微镜下一般形态学观察、透射电子显微镜超微结构观察及流式细胞术测定BCMEC凋亡变化。结果H₂O₂ (200 μmol·L^-1)损伤BCMEC后，细胞存活率下降，LDH释放增加，羟乙基葛根素和edaravone可减轻此损伤。H₂O₂ (100 μmol·L^-1)可诱导BCMEC凋亡，羟乙基葛根素 和edaravone对此有保护作用。结论羟乙基葛根素和edaravone对H₂O₂导致的BCMEC坏死和凋亡有保护作用，该作用与其抗氧化作用有关。     

基于能量代谢分析白藜芦醇对H2O2诱导SH-SY5Y细胞保护作用

目的 基于能量代谢探究白藜芦醇对H2O2诱导的人神经母细胞瘤SH-SY5Y细胞氧化损伤的保护作用。方法 选取20、10、5、1 μmol•L^-1的白藜芦醇（Res）处理SH-SY5Y细胞24 h后,加入1.2 mmol•L^-1的H₂O₂,继续培养24 h,采用MTT法测定细胞活力,流式细胞术检测细胞凋亡率,试剂盒检测糖代谢相关酶己糖激酶（HK）、磷酸果糖激酶（PFK）、丙酮酸激酶（PK）、琥珀酸脱氢酶（SDH）、乳酸脱氢酶（LDH）活力以及葡萄糖消耗量、ATP含量,Western blot法检测低氧诱导因子1α（HIF-1α）和葡萄糖转运体1（GLUT-1）表达。结果 1.2 mmol•L^-1的H₂O₂造成SH-SY5Y细胞氧化损伤,细胞活力降低,细胞凋亡率升高（P<0.01）;与H₂O₂氧化损伤组相比,20、10、5 μmol•L^-1白藜芦醇组细胞活力升高,细胞凋亡率显著降低（P<0.01）;PFK、PK、SDH活力及ATP含量、葡萄糖消耗量显著升高,HK活力和细胞外LDH活力明显降低（P<0.01）;GLUT-1表达量显著升高,HIF-1α表达量显著降低（P<0.01）。结论 20、10、5 μmol•L^-1的白藜芦醇调控GLUT-1和HIF-1α蛋白表达,提高细胞糖代谢酶活力,增加产能,对H₂O₂诱导的SH-SY5Y细胞氧化损伤发挥保护作用。     

白藜芦醇对麻醉大鼠颈动脉窦压力感受器的作用

在隔离灌流左侧颈动脉窦区的麻醉大鼠上观察了白藜芦醇对颈动脉窦压力感受器活动的影响。隔离灌流麻醉大鼠的颈动脉窦区，同时记录窦神经放电，并绘制压力感受器活动的机能曲线。白藜芦醇(30， 60及120 μmol·L^-1)隔离灌流颈动脉窦区时，压力感受器活动的机能曲线向右下方移位，曲线的斜率以及窦神经放电的最大积分值显著下降，且其变化呈一定的剂量依赖性。预先应用NO合酶抑制剂(L-NAME， 100 μmol·L^-1)可完全消除白藜芦醇对压力感受器活动的抑制作用；预先应用钙通道的开放剂(Bay K8644， 500 nmol·L^-1)可以取消白藜芦醇的抑制作用；预先应用正矾酸钠(sodium orthovanadate， 1 mmol·L^-1)后，对白藜芦醇抑制压力感受器活动的作用无影响。白藜芦醇对大鼠颈动脉窦压力感受器活动有抑制作用，此作用可能与局部NO的释放及减弱牵张敏感性通道介导的钙离子内流有关。     

Silica nanoparticles induced intrinsic apoptosis in neuroblastoma SH-SY5Y cells via CytC/Apaf-1 pathway

The present study was to investigate effects of Silica nanoparticles (SiNPs) on nervous system and explore potential mechanisms in human neuroblastoma cells (SH-SY5Y). Cytotoxicity was detected by cell viability and Lactate dehydrogenase (LDH) release. Flow cytometry analysis was applied to assess mitochondrial membrane potential (MMP) loss, intracellular Ca²⁺ and apoptosis. To clarify the mechanism of SiNPs-induced apoptosis, intrinsic apoptosis-related proteins were detected. Our results showed that SiNPs caused cytotoxicity, cell membrane damage and Ca²⁺ increase in a dose-dependent manner in SH-SY5Y cells. Both the mitochondrial membrane potential (MMP) loss and potential mitochondria damage resulted in Cyt C release to the cytoplasm. The elevated Cyt C and Apaf1 further triggered intrinsic apoptosis via executive molecular caspase-9 and caspase-3. The present study confirmed that SiNPs induced intrinsic apoptosis in neuroblastoma SH-SY5Y cells via CytC/Apaf-1 pathway and provided a better understanding of the potential toxicity induced by SiNPs on human neurocyte.     

虾青素可能通过H~+传递功能保护NaN_3损伤的人胎肝L-02细胞

观察虾青素(astaxanthin)对呼吸链复合体Ⅳ抑制剂叠氮钠(NaN3)损伤的人胎肝L-02细胞保护作用,并初步探讨其作用机制。100 mmol·L-1 NaN3用于构建肝损伤细胞模型,通过测定不同浓度虾青素(0.01、0.10、1.00及10.00 nmol·L-1)对损伤细胞存活率(MTT检测)、细胞内活性氧(reactive oxygen species,ROS)水平(DCFH-DA检测)、细胞凋亡率(Annexin V-FITC/PI双染法)以及线粒体膜电位(mitochondrial membrane potential,MMP)水平(JC-1法)的影响,发现虾青素能抑制损伤细胞晚期凋亡;对细胞存活率和MMP的保护作用呈现先增加后降低的非剂量依赖性关系,其中0.10 nmol·L-1虾青素表现为较强的保护作用;实验浓度范围内的虾青素并不能显著降低细胞内ROS水平(P>0.05)。为进一步探讨虾青素对损伤细胞的保护作用,人工制备平面双层磷脂膜(planar bilayer lipid membrane,BLM)模拟线粒体膜,测定不同浓度虾青素(0.1%、2.0%、10.0%)对H+的传递能力。结果...     

松果菊苷对TNFα诱导的SH-SY5Y细胞凋亡的保护作用

目的　探讨肉苁蓉提取物松果菊苷对TNFα诱导的SH SY5Y细胞凋亡的保护作用。方法　用MTT法检测细胞存活率,DNA琼脂糖凝胶电泳和流式细胞仪检测细胞凋亡的发生,以激光共聚焦显微镜荧光染色法检测细胞内活性氧的产生和线粒体膜电位的变化,并用荧光酶标仪测定caspase 3的活性。结果　100μg·L-1 TNFα处理细胞 36h显著降低细胞的存活率;诱导细胞发生凋亡,凋亡率达37%;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,红 /绿荧光强度的比值由正常的 5 97降低为 0 35左右。而预先给予 1, 10或者 100mg·L-1浓度的松果菊苷处理细胞 2h,可提高细胞存活率;并可有效抑制DNAladder的发生;流式细胞仪检测凋亡率分别降低到25 9%, 18 3% 和 8 2%;激光共聚焦显微镜结果显示松果菊苷可明显抑制细胞内活性氧产生;并可逐渐恢复线粒体的高能量状态;caspase 3的活性不断降低,并呈现了一定的剂量依赖性。结论　松果菊苷能抑制TNFα诱导的SH SY5Y细胞凋亡,其神经细胞保护作用可能与降低细胞内活性氧水平,抑制caspase 3的活性和维持线粒体膜电位的高能状态有关。     

Protective effects of salidroside on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells

Oxidative stress plays an important role in Alzheimer's disease and other neurodegenerative disorders. Salidroside, a phenylpropanoid glycoside isolated from Rhodiola rosea L, shows potent antioxidant property. In this paper, the neuroprotective effects of salidroside on hydrogen peroxide (H2O2)-induced apoptosis in SH-SY5Y cells were investigated. Pretreatment with salidroside markedly attenuated H2O2-induced cell viability loss and apoptotic cell death in a dose-dependent manner. The mechanisms by which salidroside protected neuron cells from oxidative stress included the induction of several antioxidant enzymes, thioredoxin, heme oxygenase-1, and peroxiredoxin-I; the downregulation of pro-apoptotic gene Bax and the upregulation of anti-apoptotic genes Bcl-2 and Bcl-X(L). Furthermore, salidroside dose-dependently restored H2O2-induced loss of mitochondrial membrane potential as well as the elevation of intracellular calcium level. These results suggest that salidroside has protective effects against oxidative stress-induced cell apoptosis, which might be a potential therapeutic agent for treating or preventing neurodegenerative diseases implicated with oxidative stress.     

PINK1/Parkin-mediated mitophagy play a protective role in manganese induced apoptosis in SH-SY5Y cells

Manganese (Mn) as an environmental risk factor of Parkinson's disease (PD) is considered to cause manganism. Mitophagy is thought to play a key role in elimination the injured mitochondria. The goal of this paper was to explore whether the PINK1/Parkin-mediated mitophagy is activated and its role in Mn-induced mitochondrial dysfunction and cell death in SH-SY5Y cells. Here, we investigated effects of MnCl₂ on ROS generation, mitochondrial membrane potential (MMP/ΔΨ_m) and apoptosis by FACS and examined PINK1/Parkin-mediated mitophagy by western-blotting and the co-localization of mitochondria and acidic lysosomes. Further, we explore the role of mitophagy in Mn-induced apoptosis by inhibition the mitophagy by knockdown Parkin level. Results show that MnCl₂ dose-dependently caused ΔΨ_m decrease, ROS generation and apoptosis of dopaminergic SH-SY5Y cells. Moreover, Mn could induce mitophagy and PINK1/Parkin-mediated pathway was activated in SH-SY5Y cells. Transient transfection of Parkin siRNA knockdown the expressing level of parkin inhibited Mn-induced mitophagy and aggravated apoptosis of SH-SY5Y cells. In conclusion, our study demonstrated that Mn may induce PINK1/Parkin-mediated mitophagy, which may exert significant neuro-protective effect against Mn-induced dopaminergic neuronal cells apoptosis.     

Protective Effect of Total Phenolic Compounds from Inula helenium on Hydrogen Peroxide-induced Oxidative Stress in SH-SY5Y Cells

Inula helenium has been reported to contain a large amount of phenolic compounds, which have shown promise in scavenging free radicals and prevention of neurodegenerative diseases. This study is to investigate the neuroprotective effects of total phenolic compounds from I. helenium on hydrogen peroxide-induced oxidative damage in human SH-SY5Y cells. Antioxidant capacity of total phenolic compounds was determined by radical scavenging activity, the level of intracellular reactive oxygen species and superoxide dismutase activity. The cytotoxicity of total phenolic compounds was determined using a cell counting kit-8 assay. The effect of total phenolic compounds on cell apoptosis due to hydrogen peroxide-induced oxidative damage was detected by Hoechst 33258 and Annexin-V/PI staining using fluorescence microscope and flow cytometry, respectively. Mitochondrial function was evaluated using the mitochondrial membrane potential and mitochondrial ATP synthesis by JC-1 dye and high performance liquid chromatography, respectively. It was shown that hydrogen peroxide significantly induced the loss of cell viability, increment of apoptosis, formation of reactive oxygen species, reduction of superoxide dismutase activity, decrease in mitochondrial membrane potential and a decrease in adenosine triphosphate production. On the other hand, total phenolic compounds dose-dependently reversed these effects. This study suggests that total phenolic compounds exert neuroprotective effects against hydrogen peroxide-induced oxidative damage via blocking reactive oxygen species production and improving mitochondrial function. The potential of total phenolic compounds and its neuroprotective mechanisms in attenuating hydrogen peroxide-induced oxidative stress-related cytotoxicity is worth further exploration.     

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.     

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.