一氧化氮对肝癌细胞线粒体膜通透性和胞浆中细胞色素C含量的影响

目的探讨一氧化氮(NO)对肝癌细胞线粒体膜通透性和胞浆中细胞色素C(cytC)含量的影响.方法用NO供体硝普钠(SNP)诱导SMMC-7721和HepG2肝癌细胞株凋亡,流式细胞术检测SMMC-7721和HepG2细胞凋亡率,MTT法观察肝癌细胞生长增殖情况,流式细胞术检测细胞线粒体跨膜电位变化,Western blot检测胞浆中cyt C含量的变化,同时应用线粒体膜通透性转变孔开放抑制剂CsA和γ-谷氨酰半胱氨酸合成酶抑制剂BSO预处理细胞,并观察以上各指标的变化.结果 SNP能诱导人肝癌细胞SMMC-7721和HepG2凋亡,并可导致两株细胞线粒体跨膜电位下降,胞浆中cyt C含量增加,与SNP的作用时间成正比.CsA能够抑制SNP所致的肝癌细胞线粒体跨膜电位的下降及胞浆中cyt C含量的增加;而BSO则可促进线粒体跨膜电位下降,cyt C从线粒体释放到胞浆.结论 NO可能通过下调线粒体跨膜电位、开放线粒体膜通透性转变孔并释放线粒体cyt C,来诱导SMMC-7721和HepG2细胞凋亡.     

线粒体在肝细胞凋亡中的作用及中药对其保护作用

肝细胞凋亡在肝纤维化中有着非常重要的作用,而许多促细胞凋亡信号作用于线粒体,能够改变线粒体通透性,继而导致其蛋白释放入胞质,由此构成线粒体介导的凋亡途径。中药及其有效成分能够从抗氧化、稳定线粒体膜流动性、抑制线粒体膜通透性转变及维持线粒体膜电位稳定等方面发挥线粒体保护作用,从而减少肝细胞的凋亡而发挥抗肝纤维化的作用。现就肝细胞凋亡的线粒体途径及中药保护进行综述。     

糙叶败酱提取物抗肿瘤作用机制研究

目的 观察糙叶败酱提取物诱导细胞凋亡过程中自由基和线粒体膜电位的变化.方法 D-101大孔吸附树脂分离糙叶败酱水提物,糙叶败酱提取物作用于S180荷瘤小鼠,观察生命延长率;透射电镜观察细胞凋亡情况,流式细胞仪检测细胞凋亡率,激光共聚焦显微镜检测细胞自由基水平和线粒体膜电位变化.结果 糙叶败酱提取物可引起S180细胞的凋亡,细胞线粒体膜电位降低,自由基水平升高.结论 糙叶败酱提取物引起S180细胞的凋亡,与含药血清对细胞内自由基水平和线粒体膜电位的影响有关.     

流式细胞技术检测线粒体通透转变孔道开放

目的 通过白藜芦醇诱导线粒体通透过渡孔道(PTP)开放模型,介绍一种使用流式细胞技术检测线粒体PTP的新方法.方法 提取大鼠肝脏线粒体,使用荧光探针NAO(nonylacridineorange)选择性的标记单个线粒体;TMRE标记检测线粒体膜电位的变化;流式细胞仪检测线粒体的侧向散射角(SSC)改变来反映白藜芦醇诱导的线粒体肿胀,即PTP的开放.结果 NAO标记线粒体结合流式细胞技术可以确定从细胞中分离线粒体的纯度;白藜芦醇处理线粒体后引起了TMRE荧光强度的降低以及SSC的减少,说明白藜芦醇可以诱导PTP开放,环孢霉素A(CsA)可以抑制白藜芦醇诱导的PTP开放.结论 流式细胞技术可以准确的检测线粒体膜电位、线粒体的肿胀程度和PTP的开放.     

线粒体膜通透性转换作用对再灌注损伤后肝细胞凋亡的影响

目的观察大鼠肝脏常温缺血再灌注后肝细胞线粒体通透性转换(mitochondrial permeability transition,MPT)作用及其与肝细胞凋亡的关系;同时观察线粒体膜通透性转换孔(mitochondrial permeability transition pore,PTP)开放抑制剂环孢素A(CsA)对MPT的抑制作用,以及对肝细胞凋亡的影响和可能的机制.方法将SD大鼠随机分为3组:假手术组、缺血再灌注组(I/R)、CsA组(I/R CsA).CsA组术前连续给予CsA 10 mg·kg-1·d-1灌胃4 d,其余组给予等量生理盐水.大鼠热缺血再灌注模型参考Nauta的方法,热缺血时间60 min,分别于再灌注0、1、6、24、72 h等不同时相收取肝组织标本,免疫组化法检测细胞质活性caspase-3;Western blot检测胞浆细胞色素C,TUNEL法检测肝细胞凋亡.结果缺血再灌注引起肝细胞线粒体发生MPT作用,细胞色素C由线粒体释放入细胞质,再灌注后0、1、6、24 h,I/R组及CsA组与假手术组相比,细胞质caspase-3的阳性程度明显增强,肝细胞凋亡明显增多(P<.01);CsA组与L/R组相比,再灌注1、6 h细胞色素C释放显著减少,再灌注1、6、24 h,caspase-3阳性程度明显下降(P<.01);再灌注后6、24、72 h,细胞凋亡明显减少(P<.05).结论缺血再灌注后肝细胞线粒体发生MPT作用可能是引起肝细胞凋亡的关键环节;CsA可能通过抑制PT孔的开放而抑制MPT作用、减少再灌注后细胞色素C释放、抑制caspase-3活化、缓解再灌注后大鼠肝细胞的凋亡.     

PIG11高表达诱导HepG2细胞凋亡及其机制的初步探讨

目的利用已建立的稳定高表达PIG11蛋白的HepG2细胞株,探讨PIG11蛋白表达对HepG2细胞凋亡的影响,以及线粒体膜电位改变和Cyt C释放在凋亡过程中的作用。阐明PIG11诱导细胞凋亡的分子机制。方法 PI染色,流式检测细胞的凋亡情况。用Rh123标记,流式测定线粒体膜电位。Western Blot检测胞浆和线粒体中Cyt C的含量变化。结果流式细胞仪检测Rh123荧光强度,结果显示：pLXSN-PIG11-HepG2细胞荧光强度（5.4±0.15）低于HepG2细胞（14.7±0.56）和pLXSN-HepG2细胞（13.2±0.53）（P〈0.01）。表明PIG11高表达诱导细胞凋亡过程中存在线粒体膜电位去极化。用CsA（10μmol/L）抑制各组细胞的线粒体通透性转移孔后,流式凋亡检测结果显示pLXSN-PIG11-HepG2细胞凋亡率明显降低。Western Blot检测发现存在线粒体Cyt C向胞浆释放。结论 PIG11高表达可诱导HepG2细胞凋亡,PIG11诱导细胞凋亡机制可能由线粒体膜电位改变及Cyt C向胞浆释放而介导。     

环孢素A对人胃癌BGC823细胞增殖与凋亡的影响

目的 研究环孢素A(CsA)对人胃癌BGC823细胞的生长增殖以及凋亡的影响.方法 MTT法检测不同浓度CsA处理24、48、72 h后对BGC823细胞增殖抑制率;流式细胞术(FCM)检测细胞周期、活性氧(ROS)含量及细胞线粒体膜电位(△Ψm)改变;Annexin-V-FITC/PI双染法检测细胞凋亡率.结果 CsA在5～20 μmol/L浓度范围内和24 ～ 72 h时间范围对BGC823细胞增殖有显著抑制作用,与药物剂量、作用时间呈现量效和时效的依赖性;FCM结果显示CsA浓度依耐性使细胞周期阻滞于G0/G1期,与对照组相比差异有统计学意义(F=33.45,P＜0.05,P＜0.01);细胞凋亡率随CsA作用浓度增加而增加;5～ 20μmol/L CsA浓度范围细胞内ROS含量显著增加(F =46.17,P＜0.01),细胞线粒体膜电位明显降低.结论 CsA对BGC823细胞有抑制增殖和诱导凋亡作用,其机制可能与阻滞细胞生长周期、细胞内ROS含量增加以及线粒体膜电位下降相关.     

莫达非尼抑制过氧化氢诱导的PC12细胞凋亡

目的:研究莫达非尼(modafinil)对过氧化氢诱导的PC12细胞凋亡的抑制作用,并对其机制进行探讨.方法:以噻唑兰(MTT)法测定PC12细胞存活率;用流式细胞术检测PC12细胞凋亡的百分率;测定细胞内罗丹明123(Rhodamine123)的荧光强度,反映细胞线粒体膜电位的改变.结果:莫达非尼能抑制过氧化氢(200 μmol/L)、硝普钠(500μmol/L)和连二亚硫酸钠(2 mmol/L)对PC12细胞的损伤.过氧化氢(200 μmol/L)24h可以诱导PC12细胞凋亡(凋亡率为32.65%).莫达非尼(15,7.5 μmol/L)显著降低凋亡细胞的百分率(凋亡百分率分别为7.95%、15.46%).并且莫达非尼可以抑制过氧化氢引起的线粒体膜电位降低.结论:莫达非尼对PC12细胞有保护作用.能抑制过氧化氢诱导的PC12细胞凋亡,这一作用可能与其抑制过氧化氢引起的线粒体膜电位降低有关.     

雷公藤甲素调控p53/SLC7A11轴诱导结肠癌细胞铁死亡

目的 探讨雷公藤甲素调控p53/SLC7A11轴诱导结肠癌细胞铁死亡的分子机制。方法 培养人结肠癌细胞系（HCT116）,采用CCK-8法检测细胞活力,EdU实验和平板克隆形成实验检测细胞增殖与克隆能力;铁死亡相关检测试剂盒测定Fe2+离子、谷胱甘肽（GSH）、丙二醛（MDA）含量,流式细胞术检测活性氧自由基（ROS）水平;JC-1荧光探针检测细胞线粒体膜电位变化;Western Blot 检测p53、SLC7A11蛋白表达水平。结果 CCK-8实验显示雷公藤甲素呈浓度梯度依赖性抑制结肠癌细胞活力,其IC50为47.82nmol/L;EdU实验和平板克隆形成实验证实雷公藤甲素可显著抑制结肠癌细胞增殖与克隆能力（P<0.05）;在雷公藤甲素干预后,铁死亡相关指标检测发现,HCT116细胞 Fe2+离子、MDA及ROS水平显著升高,GSH含量降低（P<0.05）;并在荧光显微镜下观察到HCT116细胞线粒体膜电位下降（P<0.05）。进一步通过Western Blot实验,发现HCT116细胞p53蛋白表达增加,SLC7A11蛋白表达下调（P<0.05）。结论 雷公藤甲素可通过调控p53/SLC7A11轴诱导结肠癌细胞铁死亡,抑制结肠癌细胞增殖与克隆。     

镉对人肝细胞WRL-68离体线粒体结构及功能的影响

目的 研究氯化镉(CdCl2)对离体肝细胞线粒体结构及功能的影响.方法 从培养肝细胞(WRL-68)中提取线粒体,各组分别用0、1、5、10靘oL/L CdCl2进行处理.采用分光光度法检测线粒体膜通透性转运孔(MPTP)的开放程度;电镜下观察线粒体的形态改变;罗丹明123检测线粒体膜电位(MMP)的变化;测定线粒体内ATP酶LDH、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)的活性以及丙二醛(MDA)的含量.结果 随着CdCl2浓度的增加MPTP开放程度增加,并且MPTP的开放可被环孢霉素A(CsA)所抑制;电镜观察发现,线粒体经CdCl2处理后出现不同程度的肿胀变形;CdCl:诱导MMP显著降低;同时引起SOD、GSH-Px、Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶LDH活性趋于下降,5、10靘oL/L CdCl2显著下调了各种酶的活性(P＜0.05);CdCl2处理组MDA含量升高,10靘oL/L CdCl2处理组与对照组比较有统计学意义(P＜0.05).结论 CdCl2能引起离体肝细胞线粒体结构的破坏,并致MPTP开放、MMP下降、线粒体酶活性的改变等与细胞凋亡相关过程的发生.     

雷公藤甲素可损伤肝脏分类线粒体及HL7702细胞株线粒体

Objective: To observe the impairing effects of triptolide on liver mitochondria in isolated rat-liver mitochondria and human normal liver HL7702 cell line. Methods: Rat-liver mitochondria were isolated from adult female Sprague–Dawley (SD) rats. Liver mitochondria were incubated with 0, 1.25, 2.5, 5 and 10 μmol/L triptolide for detecting mitochondrial swelling and with 0, 2.5, 5 and 10 μmol/L triptolide for mitochondrial permeability transition pore (MPTP) activity. Mitochondrial swelling was estimated by measuring the apparent absorbance change during 600 s in the mitochondrial suspensions at 520 nm with a mitochondrial swelling examining kit. The effect of triptolide on MPTP was determined with a fluorescence detection kit by detecting the fluorescence intensity at an excitation wavelength of 488 nm emitted at 527 nm. Human normal liver HL7702 cells were treated without or with 0.02, 0.1 and 0.5 μmol/L triptolide for 24 h for analyzing mitochondrial transmembrane potential (△Ψm) and reactive oxygen species (ROS). △Ψm was measured using the fluorescent probe 5,5'',6,6''-tetrachloro-1,1'',3,3''-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). ROS was measured using fluorescent probe 2'',7''-dichlorofluorescin diacetate (DCFH-DA). The cells were harvested and dyed with JC-1 and DCFH-DA, and analyzed by flow cytometry, respectively. Results: Incubation of isolated mitochondria with triptolide results in swollen mitochondria in a concentration-dependent manner. Moreover, triptolide significantly activated mitochondrial permeability transition at 5 and 10 μmol/L (P<0.05 and P<0.01). When HL7702 cells were exposed to a various concentration triptolide for 24 h, mitochondrial membrane depolarization and increase of ROS were caused by triptolide in a concentration-dependent manner. Triptolide significantly induced the mitochondrial membrane depolarization at 0.1 and 0.5 μmol/L (P<0.05 and P<0.01) and the increase of ROS at 0.1 and 0.5 μmol/L (P<0.05 and P<0.01). Conclusion: Triptolide could induce mitochondrial impairment, which may be one of the mechanisms by which hepatotoxicity occurs.     

缺血再灌注引起神经元凋亡及褪黑素抗凋亡的机制

目的 探讨模拟缺血再灌注引起神经元凋亡的途径和褪黑素(melatonin，MT)抗凋亡的作用机制。方法 用原代培养的SD乳鼠小脑颗粒细胞建立以缺氧缺糖模拟缺血再灌注模型，并加不同浓度的MT(10^-5、10^-7、10^-9mol／L)孵育。采用荧光比色法检测其线粒体跨膜电位及细胞内的活性氧；用Western blot及ELISA分别检测线粒体和胞质中的细胞色素C；比色法检测胞质中Caspase-3的活性。结果 在模拟缺血再灌注小脑颗粒细胞模型中，细胞内的活性氧在缺氧缺糖后明显增加；线粒体跨膜电位降低并随再灌注时间延长而加重；线粒体释放入胞质的细胞色素C增加，胞质的Caspase-3活性增加；MT可显著减少活性氧和抑制线粒体释放细胞色素C，抑制线粒体跨膜电位的降低，并呈一定剂量依赖性。结论 ①缺血再灌注引起的神经元凋亡部分是通过线粒体凋亡途径；②MT可通过抗氧化作用和阻止线粒体凋亡而抑制模拟缺血再灌注诱导的小脑颗粒细胞凋亡。     

RNAi沉默STAT3基因联合mTOR抑制剂rapamycin诱导BEL-7402肝癌细胞凋亡

目的：探讨PI3K/AKT/mTOR和JAK/STAT3 2条信号转导途径共同作用对肝癌细胞凋亡的影响,为肝癌基因治疗提供依据。方法：选取对数生长期BEL-7402细胞,随机分为对照组、mTOR抑制剂rapamycin(Rapa)组、阴性质粒组、阴性质粒+ Rapa组、STAT3-siRNA质粒组和STAT3-siRNA 质粒+Rapa组,应用LipofectamineTM 2000转染试剂将含有目的基因的质粒转染BEL-7402细胞,同时应用rapamycin,分别采用流式细胞术和Hoechst33258荧光染色检测细胞凋亡率和形态学的变化,JC-1 荧光染色观察线粒体膜电位(ΔΨm)变化,Western blotting法检测活性caspase-3蛋白表达水平。结果：STAT3-siRNA+Rapa组细胞凋亡率为60.22%±0.87%,明显高于其他各组(P<0.05),且细胞ΔΨm明显降低(27.28%±1.82%,P<0.05);Hoechst33258荧光染色检测,见STAT3-siRNA有大量细胞出现细胞核聚集、边缘化和核
碎裂等典型细胞凋亡形态;Western blotting检测,STAT3-siRNA+Rapa组活性caspase-3蛋白表达水平明显高于其他各组（P<0.05）。结论：RNAi沉默BEL-7402肝癌细胞STAT3基因联合rapamycin可促进BEL-7402肝癌细胞的凋亡,二者具有明显的协同作用。     

Chromium（VI）—induces Production of Reactive Oxygen Species,Change of Plasma Membrane Potential and Dissipation of Mitochondria Membrane otential in Chinese Hamster Lung Cell Cultures

OBJECTIVE: To examine whether Reactive Oxygen Species (ROS) is generated, and whether plasma membrane potential and mitochondrial membrane potential are depolarized in Chinese Hamster Lung (CHL) cell lines exposed to Cr (VI). METHODS: CHL cells were incubated with Cr(VI) at 10 mumol/L, 2.5 mumol/L, 0.65 mumol/L for 3 and 6 hours, respectively. The production of ROS was performed by using 2,7-dichlorofluorescin diacetate; The changes in plasma membrane potential were estimated using fluorescent cationic dye DiBAC4; And the changes in mitochondria membrane potential were estimated using fluorescent dye Rhodamine 123. RESULTS: The ROS levels in CHL cells increased in all treated groups compared with the control group (P < 0.01); The plasma membrane potential and mitochondrial membrane potential in CHL cells dissipated after incubated with Cr(VI) at 10 mumol/L for 3 hours and 6 hours (P < 0.01), at 2.5 mumol/L for 6 hours (P < 0.01 or 0.05). CONCLUSION: Cr(VI) causes the dissipation of plasma membrane potential and mitochondrial membrane potential in CHL cell cultures, and Cr(VI)-induced ROS may play a role in the injuries.     

Protective Effect of Ozone against Hemiscorpius lepturus Envenomation in Mice

Objective Scorpion(Hemiscorpius lepturus) stings are a public health concern in Iran, particularly in south and southwestern regions of Iran. The gold standard for the treatment of a scorpion sting is anti-venom therapy. However, immunotherapy can have serious side effects, such as anaphylactic shock(which can sometimes even lead to death). The aim of the current study was to demonstrate the protective effect of ozone against toxicity induced by Hemiscorpius lepturus(H. lepturus) venom in mice. Methods Eight hours after the injection of ozone to the experimental design groups, the male mice were decapitated and mitochondria were isolated from five different tissues(liver, kidney, heart, brain, and spinal cord) using differential ultracentrifugation. Then, assessment of mitochondrial parameters including mitochondrial reactive oxidative species(ROS) production, mitochondrial membrane potential(MMP), ATP level, and the release of cytochrome c from the mitochondria was performed. Results Our results showed that H. lepturus venom-induced oxidative stress is related to ROS production and MMP collapse, which is correlated with cytochrome c release and ATP depletion, indicating the predisposition to the cell death signaling. Conclusion In general, ozone therapy in moderate dose can be considered as clinically effective for the treatment of H. lepturus sting as a protective and antioxidant agent.     

Protective Effect of Ozone against Hemiscorpius Iepturus Envenomation in Mice

Objective Scorpion (Hemiscorpius lepturus) stings are a public health concern in Iran, particularly in south and southwestern regions of Iran. The gold standard for the treatment of a scorpion sting is anti-venom therapy. However, immunotherapy can have serious side effects, such as anaphylactic shock (which can sometimes even lead to death). The aim of the current study was to demonstrate the protective effect of ozone against toxicity induced by Hemiscorpius lepturus (H. lepturus) venom in mice. Methods Eight hours after the injection of ozone to the experimental design groups, the male mice were decapitated and mitochondria were isolated from five different tissues (liver, kidney, heart, brain, and spinal cord) using differential ultracentrifugation. Then, assessment of mitochondrial parameters including mitochondrial reactive oxidative species (ROS) production, mitochondrial membrane potential (MMP), ATP level, and the release of cytochrome c from the mitochondria was performed. Results Our results showed that H. lepturus venom-induced oxidative stress is related to ROS production and MMP collapse, which is correlated with cytochrome c release and ATP depletion, indicating the predisposition to the cell death signaling. Conclusion In general, ozone therapy in moderate dose can be considered as clinically effective for the treatment of H. lepturus sting as a protective and antioxidant agent.     

神经干细胞与大鼠RPE细胞间的线粒体转运及对RPE细胞凋亡的影响

目的 研究小鼠神经干细胞(neural stem cells, NSCs)C17.2细胞系与从RCS大鼠变性视网膜中原代分离视网膜色素上皮细胞(retinal pigment epithelial,RPE)的线粒体交换方式及其对RPE细胞特性的影响.方法 分离RCS大鼠RPE细胞并进行培养和鉴定.分别用线粒体特异性标记物Mitotracker-red和Mitotracker-green标记RPE细胞和小鼠NSCs细胞的线粒体,将两种细胞共培养,激光共聚焦显微镜下观察两种细胞之间隧道纳米管(tunneling nanotubes,TNT)的形成及线粒体转运方式.采用流式细胞仪检测与NSCs共培养后RPE细胞的反应性活性氧类(reactive oxygen species,ROS)水平、细胞周期和细胞凋亡水平的变化.结果 来源于RCS大鼠视网膜的第3代RPE细胞生长状态良好,RPE65及Bestrophin蛋白阳性率细胞均大于95%.与NSCs共培养24 h后,可见RPE细胞与NSCs间TNT的形成,NSCs中的线粒体向RPE细胞方向单向运动,接受NSCs转运的线粒体后,RPE细胞的ROS水平降低(P<0.01);RPE细胞的增殖能力增加,处于S期的细胞比例明显增加(P<0.01),细胞增殖指数(PI)显著增加(P<0.05);RPE细胞早期凋亡细胞比例显著降低(P<0.05).结论 NSCs可通过与相邻的变性RPE细胞之间形成TNT并向其转运线粒体而改善变性RPE细胞的存活.     

大鼠肝再生时线粒体通透性转换的变化

目的:研究大鼠肝再生过程中肝细胞线粒体膜通透性转换(PT)的变化规律.方法:SD大鼠肝70%部分切除(PH)制作肝再生模型,断头取肝分离线粒体后,通过检测静息态和不同浓度钙离子诱导下线粒体悬液在540 nm处光密度值(D540)变化来观察线粒体PT的时相变化.结果:与对照组比较,大鼠肝再生早期(PH后0～24 h)和后期(PH后120～168 h)都表现为肝线粒体先收缩后肿胀,也即通透性先下降后增高;同时大鼠肝再生早期肝线粒体可明显抵抗钙离子的诱导作用,而PH后24 h和168 h组的大鼠肝线粒体对钙离子的诱导非常敏感.环孢素A(CsA)可阻断钙的诱导作用.结论:大鼠肝再生过程中线粒体PT出现明显规律性改变,这可能与肝再生过程中线粒体氧化磷酸化的变化以及肝再生的启动和终止有关.     

Inhibition of Phosphodiesterase 4 by FCPR16 Protects SH-SY5Y Cells against MPP+-Induced Cell Death through Activating cAMP/PKA/CREB and Epac/Akt Signaling Pathways

Background: Phosphodiesterase 4 (PDE4) is a promising target for the treatment of Parkinson's disease (PD). However, the underlying mechanism has not yet been well elucidated. Additionally, most of current PDE4 inhibitors produce severe nausea and vomiting response in patients, which limit their clinical application. FCPR16 is a novel PDE4 inhibitor with little emetic potential. In the present study, the neuroprotective effect and underlying mechanism of FCPR16 against cellular apoptosis induced by 1-methyl-4-phenylpyridinium (MPP+) were examined in SH-SY5Y cells and primary cultured neurons. Methods: CCK-8 assay, Hoechst staining, lactate dehydrogenase release and flow cytometry were used to study the protective effect of FCPR16 against cell damage caused by MPP+. Mitochondrial membrane potential (Δψm) was measured by JC-1 staining. The extent of oxidation was evaluated using Cell ROXs Deep Red Reagent and malonaldehyde (MDA) kit. Pretreatments with various pathway inhibitors were used to investigate the possible pathways involved in the protection of FCPR16. The phosphorylated and total levels of various proteins were analyzed by Western blot. Results: FCPR16 (12.5–50 μM) dose-dependently reduced MPP+-induced loss of cell viability, accompanied by reductions in nuclear condensation and lactate dehydrogenase release. The level of cleaved caspase 3 and the ratio of Bax/Bcl-2 were also decreased after treatment with FCPR16 in MPP+-treated cells. Furthermore, FCPR16 (25 μM) significantly suppressed the accumulation of reactive oxygen species (ROS), prevented the decline of Δψm and attenuated the expression of MDA level. Further studies disclosed that FCPR16 enhanced the levels of cAMP and the exchange protein directly activated by cAMP (Epac) in SH-SY5Y cells. Western blotting analysis revealed that FCPR16 increased the phosphorylation of cAMP response element-binding protein (CREB) and protein kinase B (Akt) down-regulated by MPP+ in SH-SY5Y cells. Moreover, the inhibitory effects of FCPR16 on the production of ROS and Δψm loss could be blocked by PKA inhibitor H-89 and Akt inhibitor KRX-0401. We also found that MPP+ induced a dose-dependent apoptosis in cultured neurons, and 500 μM MPP+ caused an approximately 50% loss of cortical neurons, while treatment with FCPR16 reversed the toxic effect of MPP+ and enhanced the cell viability in a dose-dependent manner. Conclusions: These results suggest that FCPR16 attenuates MPP+-induced dopaminergic degeneration via lowering ROS and preventing the loss of Δψm in SH-SY5Y cells. Mechanistically, cAMP/PKA/CREB and Epac/Akt signaling pathways are involved in these processes.