HPVm16E7刺激树突状细胞SOCS1基因沉默后对宫颈癌小鼠模型免疫效应的影响

摘要 目的：在宫颈癌细胞TC-1的荷瘤小鼠模型中探讨树突状细胞（DCs）的细胞因子信号抑制物1（SOCS1）基因沉默后对宫颈癌细胞免疫效应的影响。方法：构建宫颈癌细胞系TC-1的荷瘤小鼠模型,将含有SOCS1沉默基因的慢病毒载体感染DCs细胞,对荷瘤小鼠进行细胞免疫后监测小鼠体内肿瘤生长、小鼠存活率,并检测小鼠脾细胞对TC-1细胞的体外裂解率以及γ干扰素（IFN-γ）、白细胞介素-12（IL-12）表达等指标。结果：DCs SOCS1基因沉默后可延长小鼠存活期,增强DCs对小鼠体内肿瘤生长的抑制作用,增强小鼠脾细胞对TC-1细胞的体外裂解率(P<0.05),增加小鼠血清IL-12因子表达(P<0.05)和小鼠脾细胞IFN-γ表达(P<0.05),但对小鼠脾内效应细胞的数量没有影响(P>0.05)。结论：小鼠体内实验初步证实,DCs SOCS1基因沉默后可一定程度上增强小鼠体内效应细胞对肿瘤细胞的杀伤效果。     

一种羊肚菌源多肽MIP-16的分离纯化及其神经保护活性

分离纯化人工栽培的梯棱羊肚菌子实体多肽(MIP-16),对其结构和神经保护活性进行研究.采用磷酸盐缓冲液提取,分子排阻色谱分离,反相高效液相色谱纯化获得梯棱羊肚菌子实体多肽,通过液相色谱-质谱连用技术完成氨基酸序列鉴定.构建6-羟基多巴胺处理后引起凋亡的PC12细胞模型,验证MIP-16对PC12细胞的保护作用.结果 ...     

罗布麻叶总黄酮提取物促进小鼠抗抑郁基因CREB、BDNF的表达

罗布麻是中国药典收录的传统中药,本研究提取它的主要有效成分罗布麻总黄酮,采用经典抗抑郁评价模型小鼠强迫游泳实验对罗布麻叶总黄酮进行抗抑郁活性评价.试验中给小鼠罗布麻叶总黄酮提取物25 mg/kg、50 mg/kg和100 mg/kg后,观察罗布麻叶总黄酮对小鼠强迫游泳不动时间的影响,结果显示罗布麻总黄酮提取物能显著缩短小鼠强迫游泳不动时间(P<0.05),药效与氟西汀相似,表明罗布麻具有明显的抗抑郁活性.在此基础上,应用皮质酮损伤的PC12细胞模型,采用荧光相对定量RT-PCR法检测罗布麻对皮质酮损伤的PC12细胞中脑内脑源性神经营养因子(BDNF)和环磷酸腺苷反应元件结合蛋白(CREB)表达水平的影响.结果表明,皮质酮处理后PC12细胞中BDNF、CREB基因的表达量最低,罗布麻处理后,表达量显著增加,较处理前增加了近十倍,且呈剂量依赖性.本研究结果提示罗布麻抗抑郁机制可能是通过(AC-cAMP-CREB)信号通路促进BDNF、CREB的基因表达而发挥的.     

橘络活性成分小分子果胶治疗非小细胞肺癌作用及机制研究北大核心CSCD

本文观察橘络活性成分小分子果胶在离体细胞水平及整体动物模型中对非小细胞肺癌的治疗作用并初步探索其作用机制。小分子果胶处理人非小细胞肺癌PC-9细胞72 h后,MTS法检测细胞活率,流式细胞仪检测细胞凋亡率;以PC-9细胞裸小鼠皮下移植瘤模型观察小分子果胶体内对肿瘤生长的抑制作用,免疫组化法检测肿瘤组织Ki67表达水平,TUNEL法检测组织中肿瘤细胞凋亡程度。结果表明小分子果胶(1.5~3 mg/m L)可显著抑制PC-9细胞增殖,与对照组相比细胞凋亡率显著升高,且具有浓度依赖性;小分子果胶(56.25 mg/kg)可显著抑制非小细胞肺癌小鼠体内肿瘤生长,肿瘤组织中细胞凋亡率显著升高。研究证明小分子果胶可显著抑制非小细胞肺癌小鼠体内肿瘤生长,该作用可能与小分子果胶抑制肿瘤细胞增殖、诱导细胞凋亡有关。     

人脐血CD34+细胞来源的树突状细胞疫苗在SCID小鼠体内的抗肿瘤免疫作用

目的观察脐血CD34 干细胞来源的树突状细胞(dendritic cells, DC)疫苗在严重联合免疫缺陷(severecombined immunity deficiency,SCID)小鼠体内对人肝癌细胞的免疫治疗和免疫保护作用.方法采用微磁珠分选系统(Mini MACS)从脐血单个核细胞中分离CD34 干细胞.重组人粒细胞-巨噬细胞集落刺激因子(recombined human granulocyte-macrophage colony-stimulating factor, rhGM-CSF)、重组人肿瘤坏死因子(recombined human tumor necrosis factor,TNF)-α诱导脐血CD34 细胞向DC分化,相差显微镜下观察DC分化过程中细胞的形态及数量变化.用人肝癌细胞BEL-7402裂解物冲击DC制备DC疫苗.在SCID小鼠体内观察DC疫苗致敏的淋巴细胞对人肝癌细胞的免疫治疗和免疫保护作用.结果脐血CD34 细胞在细胞因子诱导下,细胞形态由小变大、由圆形逐渐变为不规则形;细胞分裂扩增过程中,数量逐渐增多,形成细胞集落.经过细胞因子联合诱导2周的DC胞质突起丰富,具有典型的树枝状形态.在体内的抗肿瘤实验中,经DC疫苗致敏的人外周血淋巴细胞治疗荷瘤小鼠,肿瘤生长速度、瘤体积和重量明显小于未致敏淋巴细胞治疗组(P<0.05).与未致敏淋巴细胞免疫组和DC疫苗致敏的淋巴细胞治疗组比较,经DC疫苗致敏的淋巴细胞免疫SCID小鼠,肝癌细胞攻击后,肿瘤的发病率降低(P<0.05),发病潜伏期延长(P<0.01),肿瘤体积和重量减小(P<0.05).结论人脐血CD34 细胞来源的DC疫苗能在一定程度上抑制人肝癌细胞在小鼠体内的生长,抵抗肿瘤细胞的攻击,对机体具有相应的抗肿瘤免疫治疗和免疫保护作用.     

人胎肝中低分子天然抑瘤物对小鼠肉瘤S—180作用的研究

从人胎儿肝脏中分离出一种分子量较小的抑瘤物质(FLS-MeOH),观察了它对小鼠肉瘤S-180细胞生长的抑制效应。在体外琼脂培养中,当FLS-MeOH浓度达到300μg/ml时,可完全抑制S-180细胞的集落形成;给荷瘤小鼠每日注射FLS-MeOH8mg/g体重,共20d,可明显延长小鼠的存活时间,部分小鼠可以无病存活。这些结果说明FLSMeOH在体内外均有明显的抗肿瘤活性,而且具有分子量小、无组织和种系特异性,以及对肿瘤细胞不可逆的毒性作用等特性。     

橘络活性成分小分子果胶治疗非小细胞肺癌作用及机制研究

本文观察橘络活性成分小分子果胶在离体细胞水平及整体动物模型中对非小细胞肺癌的治疗作用并初步探索其作用机制。小分子果胶处理人非小细胞肺癌PC-9细胞72 h后,MTS法检测细胞活率,流式细胞仪检测细胞凋亡率;以PC-9细胞裸小鼠皮下移植瘤模型观察小分子果胶体内对肿瘤生长的抑制作用,免疫组化法检测肿瘤组织Ki67表达水平,TUNEL法检测组织中肿瘤细胞凋亡程度。结果表明小分子果胶(1.5~3 mg/m L)可显著抑制PC-9细胞增殖,与对照组相比细胞凋亡率显著升高,且具有浓度依赖性;小分子果胶(56.25 mg/kg)可显著抑制非小细胞肺癌小鼠体内肿瘤生长,肿瘤组织中细胞凋亡率显著升高。研究证明小分子果胶可显著抑制非小细胞肺癌小鼠体内肿瘤生长,该作用可能与小分子果胶抑制肿瘤细胞增殖、诱导细胞凋亡有关。     

过表达Nogo-C对PC12细胞存活及增殖的影响

以PC12细胞为神经元细胞模型,研究Nogo-C对神经元细胞存活及增殖的作用。在PC12细胞中转染过表达Nogo-C,使用G418药物筛选以获得稳定表达的细胞克隆,利用Hoechst33342染色、细胞计数、MTT以及流式细胞仪等技术检测Nogo-C对细胞增殖以及细胞周期的影响。结果表明:(1)Hoechst33342染色未观察到表达Nogo-C的细胞发生明显凋亡;(2)细胞计数及MTT实验观察到转染Nogo-C后的PC12细胞生长增殖活性明显降低;(3)流式细胞仪检测细胞生长周期,正常PC12细胞G1期的百分数为(37.8±7.9)%,S期为(50.4±8.5)%,而转染Nogo-C的PC12细胞G1期为(76.8±4.1)%,S期为(14.7±1.7)%,提示转染Nogo-C的PC12细胞的细胞周期被阻滞在G1期;(4)没有获得稳定表达Nogo-C的PC12细胞模型。实验证明,过表达Nogo-C通过使PC12细胞周期被阻滞在G1期而明显抑制细胞的增殖,但是并不引起细胞的凋亡。     

小鼠去细胞拟胚体对小鼠Lewis肺癌细胞体内生长的影响

目的:阐明小鼠去细胞拟胚体对小鼠Lewis肺癌细胞在体内生长的影响。方法:先制备来源于小鼠胚胎干细胞的拟胚体,然后用SDS去细胞处理。实验分成3组:小鼠Lewis肺癌细胞与去细胞拟胚体培养组,癌细胞与Matrigel培养组和单纯癌细胞组(每组n=12)。培养3天后注射入裸鼠体内,观察肿瘤生长情况。28天取出瘤体,Ki67和CD31免疫组化染色(n=12)检测细胞增殖和肿瘤微血管密度(MVD),Western blot检测组织Paxillin,E-cadherin和β-actin水平(n=6)。结果:去细胞拟胚体组肿瘤生长明显较单纯细胞组和Matrigel组慢。去细胞拟胚体组Ki67指数((17.1±2.6)%)明显小于单纯细胞组((34.5±4.7)%)和Matrigel组((48.4±8.6)%)(P0.05);去细胞拟胚体组的MVD(18.7±3.6个/mm2)明显小于单纯细胞组(32.1±6.4个/mm2)和Matrigel组(42.6±7.1个/mm2)(P0.05)。Western blot结果提示去细胞拟胚体组的Paxillin表达小于单纯细胞组和Matrigel组(P0.05),而E-cadherin表达大于单纯细胞组和Matrigel组(P0.05)。结论:小鼠去细胞拟胚体对小鼠Lewis肺癌细胞在体内有明显的促分化作用。     

广西眼镜蛇毒神经生长因子分离纯化方法

为了得到更高纯度和活性的广西眼镜蛇毒神经生长因子(never growth factor,NGF),我们对原有的分离纯化方法进行改进。采用DEAE CL-6B离子交换柱、Sephadex G-50凝胶层析柱、Macro-prep High S离子交换柱结合的方法进行分离。在分离纯化过程中,采用PC12细胞检验每一步所得结果中的NGF活性,并测定NGF诱导PC12细胞分化的最小浓度。经DEAE CL-6B离子交换柱、Sephadex G-50凝胶层析柱、Macro-prep High S离子交换柱分离纯化后,得到的第二峰具有NGF活性,并且已达到电泳纯。经PC12细胞验证后,确定NGF诱导PC12细胞分化的最小浓度为0.1μg/m L。     

RUNX1对氧糖剥夺诱导PC12细胞凋亡的保护作用研究

目的:研究RUNX1在PC12细胞氧糖剥夺模型中的表达及其对PC12细胞的保护作用,并探讨其相关机制。方法:体外培养PC12细胞并构建氧糖剥夺模型,将细胞分为对照组、氧糖剥夺组、RUNX1 si RNA处理组、si RNA对照处理组(sicontrol)、pc DNA3.1-RUNX1处理组(pc RUNX1)和pc DNA3.1对照处理组(pc DNA 3.1)。q RT-PCR和western blot检测RUNX1、磷酸化Akt(p-Akt)和总Akt(t-Akt)表达水平;MTT法检测细胞存活率;Annexin V-FITC/PI双染法检测细胞凋亡。结果:与对照组比较,RUNX1在PC12细胞氧糖剥夺模型中表达水平显著升高;沉默RUNX1可下调PC12细胞的存活率,促进细胞的凋亡,有效抑制p-Akt蛋白表达,而过表达RUNX1显著提高细胞存活率,抑制细胞凋亡,并上调p-Akt蛋白表达;此外,PI3K/Akt通路抑制剂LY294002明显抑制RUNX1过表达对细胞存活率的促进作用和对细胞凋亡的抑制作用。结论:RUNX1可通过PI3K/Akt信号通路保护OGD对PC12细胞的损伤作用。     

Low-Dose Levodopa Protects Nerve Cells from Oxidative Stress and Up-Regulates Expression of pCREB and CD39

Objective

This study aimed to investigate the influence of low-dose levodopa (L-DOPA) on neuronal cell death under oxidative stress.

Methods

PC12 cells were treated with L-DOPA at different concentrations. We detected the L-DOPA induced reactive oxygen species (ROS). Meanwhile, MTT and LDH assay were performed to determine the proliferation and growth of PC12 cells with or without ROS scavenger. In addition, after pretreatment with L-DOPA at different concentrations alone or in combination with CD39 inhibitor, PC12 cells were incubated with hydrogen peroxide (H₂O₂) and the cell viability was evaluated by MTT and LDH assay. In addition, the expression of pCREB and CD39 was detected by immunofluorescence staining and Western blot assay in both cells and rat’s brain after L-DOPA treatment.

Results

After treatment with L-DOPA for 3 days, the cell proliferation and growth were promoted when the L-DOPA concentration was <30 µM, while cell proliferation was comparable to that in control group when the L-DOPA concentration was >30 µM. Low dose L-DOPA could protect the PC12 cells from H₂O₂ induced oxidative stress, which was compromised by CD39 inhibitor. In addition, the expression of CD39 and pCREB increased in both PC12 cells and rats’ brain after L-DOPA treatment.

Conclusions

L-DOPA at different concentrations has distinct influence on proliferation and growth of PC12 cells, and low dose (<30 µM) L-DOPA protects PC12 cells against oxidative stress which might be related to the up-regulation of CD39 and pCREB expression.     

miR-1298表达对PC-12细胞糖氧剥夺/复氧损伤的影响及其机制研究

摘要 目的：通过体外细胞培养探讨miR-1298对缺血缺氧性神经损伤的调节作用。方法：首先通过细胞活性检测和乳酸脱氢酶（LDH）细胞毒性法确定大鼠PC-12细胞糖氧剥夺/复氧（OGD/R）的造模效果,同时采用实时荧光定量PCR（RT-qPCR）检测细胞miR-1298的表达差异。体外转染miR-1298mimic、mimic NC、miR-1298 inhibitor和inhibitor NC至大鼠PC-12细胞系,检测mimic、mimic NC、inhibitor、inhibitor NC的转染效率。经过OGD/R处理后将细胞分为Control组、OGD/R组、mimic组、mimicNC组、inhibitor组和inhibitorNC组。流式细胞术检测各组PC-12细胞凋亡的情况,免疫印迹试验（Western blot）检测各组PC-12细胞凋亡相关蛋白B淋巴细胞瘤-2基因（BCL-2）和Bcl-2相关的x基因（Bax）表达的情况。结果：PC12细胞经过OGD/R处理后,其细胞存活率与Control组比明显下降且LDH漏出率明显上升（均P<0.05）;模型细胞中miR-1298相对表达量明显低于Control组（P<0.05）。转染24小时后mimic组细胞中miR-1298的相对表达量明显高于mimicNC组（P<0.05）;mimic组细胞凋亡率低于mimicNC组,而inhibitor组细胞凋亡率高于inhibitor NC组（均P<0.05）;mimic组的BCL-2表达量较mimicNC组升高,而BAX表达量下降,inhibitor组与inhibitorNC组相比,BCL-2表达量下降,而BAX表达量上升,差异均有统计学意义（均P<0.05）。结论：miR-1298通过抑制细胞凋亡减轻PC-12细胞OGD/R的损伤。     

Cucurbitacin B induces neurogenesis in PC12 cells and protects memory in APP/PS1 mice

Cucurbitacin B (CuB) isolated from Cucumis melo by using a PC12 cell bioassay system exhibited significant nerve growth factor (NGF)‐mimic or NGF‐enhancing activity in PC12 and primary neuron cells. It was also demonstrated pro‐neurogenesis effects in ICR and APP/PS1 mice and improved memory deficit of APP/PS1 mice. Its possible mechanism includes significant induction of the phosphorylation of glucocorticoid receptor (GR), protein kinase C (PKC), phospholipase C (PLC) and inhibition of cofilin. ChemProteoBase profiling, binding assay and cellular thermal shift assay (CETSA) were used to determine the target protein. Results revealed that CuB could affect actin dynamics as an actin inhibitor but did not bind with GR. The protein level of cofilin in PC12 cells after treating 0.3 μM and different temperatures was significantly higher than that of control group. Other neurotrophic signalling pathways, such as TrkA/TrkB, were analysed with specific inhibitors and Western blot. The inhibitors of TrkA, PLC, PKC, Ras, Raf and ERK1/2 significantly decreased the percentage of PC12 cells with neurite outgrowth and shortened the length of neurite outgrowth induced by CuB. CuB significantly induced the phosphorylation of TrkA, ERK and CREB. The phosphorylation of these proteins was obviously decreased by their specific inhibitors. These results suggest that cofilin is a candidate target protein of CuB in PC12 cells and that the GR/PLC/PKC and TrkA/Ras/Raf/ERK signalling pathways play important roles in the neuroprotective effect of CuB.     

Ionic responses and growth stimulation induced by nerve growth factor and epidermal growth factor in rat pheochromocytoma (PC12) cells

Rat pheochromocytoma cells (clone PC12) respond to nerve growth factor (NGF) by the acquirement of a phenotype resembling neuronal cells. In an earlier study we showed that NGF causes an increase in Na+,K+ pump activity, as monitored by ouabain-sensitive Rb+ influx. Here we show that addition of epidermal growth factor (EGF) to PC12 cells resulted in a stimulation of Na+,K+ pump activity as well. The increase of Na+,K+ pump activity by NGF or EGF was due to increased Na+ influx. This increased Na+ influx was sensitive to amiloride, an inhibitor of Na+,H+ exchange. Furthermore, no changes in membrane potential were observed upon addition of NGF or EGF. Amiloride-sensitive Na+,H+ exchange in PC12 cells was demonstrated by H+ efflux measurements and the effects of weak acids on Na+ influx. These observations suggest that both NGF and EGF activate an amiloride-sensitive, electroneutral Na+,H+ exchange mechanism in PC12 cells. These findings were surprising in view of the opposite ultimate biological effects of NGF and EGF, e.g., growth arrest vs. growth stimulation. However, within 24 h after addition, NGF was found to stimulate growth of PC12 cells, comparable to EGF. In the presence of amiloride, this stimulated growth by NGF and EGF was abolished. In contrast, amiloride did not affect NGF-induced neurite outgrowth of PC12 cells. From these observations it is concluded that in PC12 cells: (a) NGF has an initial growth stimulating effect; (b) neurite outgrowth is independent of increased amiloride-sensitive Na+ influx; and (c) growth stimulation by NGF and EGF is associated with increased amiloride-sensitive Na+ influx.     

皮质酮诱导的PC12细胞梯度应激损伤模型的构建及评价

目的:建立皮质酮诱导的PC12细胞梯度应激损伤模型,为细胞应激水平的评估和细胞应激损伤调控研究提供实验基础和对象。方法:通过检测不同浓度皮质酮(0～1 000μmol/L)在经过不同干预时间(8～48 h)后PC12细胞活力,观察皮质酮对细胞活力的影响,筛选最佳干预条件的细胞模型。分光光度法和微量法检测细胞模型的关键应激指标(MDA、SOD、NADH、LDH),对模型进行评价。结果:当皮质酮浓度在200μmol/L以下且干预时间为12 h时,细胞活力在半数失活率以下,可减少各组由于细胞活力下降而产生的混杂因素。与空白对照组比较,皮质酮浓度依赖性地升高模型组的MDA、NADH和LDH水平,降低SOD水平(P<0.01),符合梯度应激模型的构建要求。结论:成功建立了PC12细胞梯度应激损伤模型,在干预时间为12 h的情况下,干预浓度为0μmol/L、25μmol/L、50μmol/L、100μmol/L、150μmol/L、200μmol/L,使得细胞模型应激损伤程度梯度增加,可作为开展细胞应激损伤评估及调控实验的基础和对象。     

Artepillin C Derived from Propolis Induces Neurite Outgrowth in PC12m3 Cells via ERK and p38 MAPK Pathways

We investigated whether artepillin C, a major component of Brazilian propolis, acts as a neurotrophic-like factor in rat PC12m3 cells, in which nerve growth factor (NGF)-induced neurite outgrowth is impaired. When cultures of PC12m3 cells were treated with artepillin C at a concentration of 20 μM, the frequency of neurite outgrowth induced by artepillin C was approximately 7-fold greater than that induced by NGF alone. Artepillin C induced-neurite outgrowth of PC12m3 cells was inhibited by the ERK inhibitor U0126 and by the p38 MAPK inhibitor SB203580. Although artepillin C-induced p38 MAPK activity was detected in PC12m3 cells, phosphorylation of ERK induced by artepillin C was not observed. On the other hand, artepillin C caused rapid activation of ERK and the time course of the activation was similar to that induced by NGF treatment in PC12 parental cells. However, NGF-induced neurite outgrowth was inhibited by artepillin C treatment. Interestingly, inhibition of ERK by U0126 completely prevented artepillin C-induced p38 MAPK phosphorylation of PC12m3 cells. These findings suggest that artepillin C-induced activation of p38 MAPK through the ERK signaling pathway is responsible for the neurite outgrowth of PC12m3 cells.     

GTK, a Src-related tyrosine kinase, induces nerve growth factor-independent neurite outgrowth in PC12 cells through activation of the Rap1 pathway. Relationship to Shb tyrosine phosphorylation and elevated levels of focal adhesion kinase

The rat pheochromocytoma cell line PC12 is extensively used as a model for studies of neuronal cell differentiation. These cells develop a sympathetic neuron-like phenotype when cultured in the presence of nerve growth factor. The present study was performed in order to assess the role of mouse GTK (previously named BSK/IYK), a cytoplasmic tyrosine kinase belonging to the Src family, for neurite outgrowth in PC12 cells. We report that PC12 cells stably overexpressing GTK exhibit a larger fraction of cells with neurites as compared with control cells, and this response is not accompanied by an increased ERK activity. Treatment of the cells with the MEK inhibitor PD98059 did not reduce the GTK-dependent increased in neurite outgrowth. GTK expression induces a nerve growth factor-independent Rap1 activation, probably through altered CrkII signaling. We observe increased CrkII complex formation with p130(Cas), focal adhesion kinase (FAK), and Shb in PC12-GTK cells. The expression of GTK also correlates with a markedly increased content of FAK, phosphorylation of the adaptor protein Shb, and an association between these two proteins. Transient transfection of GTK-overexpressing cells with RalGDS-RBD or Rap1GAP, inhibitors of the Rap1 pathway, reduces the GTK-dependent neurite outgrowth. These data suggest that GTK participates in a signaling pathway, perhaps involving Shb, FAK and Rap1, that induces neurite outgrowth in PC12 cells.     

Protective Effects of YC-1 Against Glutamate Induced PC12 Cell Apoptosis

Glutamate, one of the major neurotransmitters in the central nervous system, is released into the synaptic spaces and bound to the glutamate receptors which facilitate normal synaptic transmission, synaptic plasticity, and brain development. Past studies have shown that glutamate with high concentration is a potent neurotoxin capable of destroying neurons through many signal pathways. In this research, our main purpose was to determine whether the specific soluble guanylyl cyclase activator YC-1 (3-(5′-hydroxymethyl-2′-furyl)-1-benzyl indazole) had effect on glutamate-induced apoptosis in cultured PC12 cells. The differentiated PC12 cells impaired by glutamate were used as the cell model of excitability, and were exposed to YC-1 or/and ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one) with gradient concentrations for 24 h. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl) assay was used to detect the cellular viability. Radioimmunoassay (RIA) was used to detect the cGMP (cyclic guanosine monophosphate) concentrations in PC12 cells. Hoechst 33258 staining and flow cytometric analysis were used to detect the cell apoptosis. The cellular viability was decreased and the apoptotic rate was increased when PC12 cells were treated with glutamate. Cells treated with YC-1 or/and ODQ showed no significant differences in the cell viability and intracellular cGMP levels compared with those of control group. The specific soluble guanylyl cyclase (sGC) inhibitor ODQ showed an inhibitory effect on cGMP level and aggravated the apoptosis of PC12 cells induced by glutamate. YC-1 elevated cGMP level thus decreased PC12 cell apoptosis induced by glutamate, but this effect could be reversed by ODQ. These results revealed that YC-1 might attenuate glutamate-induced PC12 cell apoptosis via a sGC–cGMP involved pathway.