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Ligustilide protects PC12 cells from oxygen-glucose deprivation/reoxygenation-induced apoptosis via the LKB1-AMPK-mTOR signaling pathway 预览
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作者 Dan-Yang Zhao Dong-Dong Yu +1 位作者 Li Ren Guo-Rong Bi 《中国神经再生研究:英文版》 SCIE CAS CSCD 2020年第3期473-481,共9页
Autophagy has been shown to have a protective effect against brain damage.Ligustilide(LIG)is a bioactive substance isolated from Ligusticum chuanxiong,a traditional Chinese medicine.LIG has a neuroprotective effect;ho... Autophagy has been shown to have a protective effect against brain damage.Ligustilide(LIG)is a bioactive substance isolated from Ligusticum chuanxiong,a traditional Chinese medicine.LIG has a neuroprotective effect;however,it is unclear whether this neuroprotective effect involves autophagy.In this study,PC12 cells were treated with 1×10^-5–1×10^-9 M LIG for 0,3,12 or 24 hours,and cell proliferation was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)assay.Treatment with 1×10^-6 M LIG for 3 hours had the greatest effect on cell proliferation,and was therefore used for subsequent experiments.PC12 cells were pre-treated with 1×10^-6 M LIG for 3 hours,cultured in 95%N2/5%CO2 in Dulbecco’s modified Eagle’s medium without glucose or serum for 4 hours,and then cultured normally for 16 hours,to simulate oxygen-glucose deprivation/reoxygenation(OGD/R).Cell proliferation was assessed with the MTS assay.Apoptosis was detected by flow cytometry.The expression levels of apoptosis-related proteins,Bcl-2 and Bax,autophagy-related proteins,Beclin 1 and microtubule-associated protein l light chain 3B(LC3-II),and liver kinase B1(LKB1)-5′-adenosine monophosphate-activated protein kinase(AMPK)-mammalian target of rapamycin(mTOR)signaling pathway-related proteins were assessed by western blot assay.Immunofluorescence staining was used to detect LC3-II expression.Autophagosome formation was observed by electron microscopy.LIG significantly decreased apoptosis,increased Bcl-2,Beclin 1 and LC3-II expression,decreased Bax expression,increased LC3-II immunoreactivity and the number of autophagosomes,and activated the LKB1-AMPK-mTOR signaling pathway in PC12 cells exposed to OGD/R.The addition of the autophagy inhibitor 3-methyladenine or dorsomorphin before OGD/R attenuated the activation of the LKB1-AMPK-mTOR signaling pathway in cells treated with LIG.Taken together,our findings show that LIG promotes autophagy and protects PC12 cells from apoptosis induced by 展开更多
关键词 AMPK apoptosis autophagy Bax Bcl-2 BECLIN 1 LC3-II LIGUSTILIDE mTOR PC12 cells
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Protective effects of organic extracts of Alpinia oxyphylla against hydrogen peroxide-induced cytotoxicity in PC12 cells 预览
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作者 Li-Hong Duan Meng Li +10 位作者 Chun-Bao Wang Qing-Mei Wang Quan-Quan Liu Wan-Feng Shang Ya-Jin Shen Zhuo-Hua Lin Tong-Yang Sun Zheng-Zhi Wu Ying-Hong Li Yu-Long Wang Xun Luo 《中国神经再生研究:英文版》 SCIE CAS CSCD 2020年第4期682-689,共8页
Alpinia oxyphylla,a traditional herb,is widely used for its neuroprotective,antioxidant and memory-improving effects.However,the neuroprotective mechanisms of action of its active ingredients are unclear.In this study... Alpinia oxyphylla,a traditional herb,is widely used for its neuroprotective,antioxidant and memory-improving effects.However,the neuroprotective mechanisms of action of its active ingredients are unclear.In this study,we investigated the neuroprotective effects of various organic extracts of Alpinia oxyphylla on PC12 cells exposed to hydrogen peroxide-induced oxidative injury in vitro.Alpinia oxyphylla was extracted three times with 95%ethanol(representing extracts 1–3).The third 95%ethanol extract was dried and resuspended in water,and then extracted successively with petroleum ether,ethyl acetate and n-butanol(representing extracts 4–6).The cell counting kit-8 assay and microscopy were used to evaluate cell viability and observe the morphology of PC12 cells.The protective effect of the three ethanol extracts(at tested concentrations of 50,100 and 200μg/mL)against cytotoxicity to PC12 cells increased in a concentration-dependent manner.The ethyl acetate,petroleum ether and n-butanol extracts(each tested at 100,150 and 200μg/mL)had neuroprotective effects as well.The optimum effective concentration ranged from 50–200μg/mL,and the protective effect of the ethyl acetate extract was comparatively robust.These results demonstrate that organic extracts of Alpinia oxyphylla protect PC12 cells against apoptosis induced by hydrogen peroxide.Our findings should help identify the bioactive neuroprotective components in Alpinia oxyphylla. 展开更多
关键词 active INGREDIENTS ALPINIA oxyphylla apoptosis ethanol crude extract fraction hydrogen PEROXIDE nerve regeneration NEUROPROTECTIVE agent NEUROPROTECTIVE effects PC12 cells traditional HERB
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Transfer of mitochondria from mesenchymal stem cells derived from induced pluripotent stem cells attenuates hypoxia-ischemia-induced mitochondrial dysfunction in PC12 cells 预览
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作者 Yan Yang Gen Ye +5 位作者 Yue-Lin Zhang Hai-Wei He Bao-Qi Yu Yi-Mei Hong Wei You Xin Li 《中国神经再生研究:英文版》 SCIE CAS CSCD 2020年第3期464-472,共9页
Mitochondrial dysfunction in neurons has been implicated in hypoxia-ischemia-induced brain injury.Although mesenchymal stem cell therapy has emerged as a novel treatment for this pathology,the mechanisms are not fully... Mitochondrial dysfunction in neurons has been implicated in hypoxia-ischemia-induced brain injury.Although mesenchymal stem cell therapy has emerged as a novel treatment for this pathology,the mechanisms are not fully understood.To address this issue,we first co-cultured 1.5×10^5 PC12 cells with mesenchymal stem cells that were derived from induced pluripotent stem cells at a ratio of 1:1,and then intervened with cobalt chloride(CoCl2)for 24 hours.Reactive oxygen species in PC12 cells was measured by Mito-sox.Mitochondrial membrane potential(ΔΨm)in PC12 cells was determined by JC-1 staining.Apoptosis of PC12 cells was detected by terminal deoxynucleotidal transferase-mediated dUTP nick end-labeling staining.Mitochondrial morphology in PC12 cells was examined by transmission electron microscopy.Transfer of mitochondria from the mesenchymal stem cells derived from induced pluripotent stem cells to damaged PC12 cells was measured by flow cytometry.Mesenchymal stem cells were induced from pluripotent stem cells by lentivirus infection containing green fluorescent protein in mitochondria.Then they were co-cultured with PC12 cells in Transwell chambers and treated with CoCl2 for 24 hours to detect adenosine triphosphate level in PC12 cells.CoCl2-induced PC12 cell damage was dose-dependent.Co-culture with mesenchymal stem cells significantly reduced apoptosis and restoredΔΨm in the injured PC12 cells under CoCl2 challenge.Co-culture with mesenchymal stem cells ameliorated mitochondrial swelling,the disappearance of cristae,and chromatin margination in the injured PC12 cells.After direct co-culture,mitochondrial transfer from the mesenchymal stem cells stem cells to PC12 cells was detected via formed tunneling nanotubes between these two types of cells.The transfer efficiency was greatly enhanced in the presence of CoCl2.More importantly,inhibition of tunneling nanotubes partially abrogated the beneficial effects of mesenchymal stem cells on CoCl2-induced PC12 cell injury.Mesenchymal stem cells reduced CoCl2-induced 展开更多
关键词 apoptosis brain injury HYPOXIA-ISCHEMIA INDUCED pluripotent STEM CELLS mesenchymal STEM CELLS MITOCHONDRIAL membrane potential MITOCHONDRIAL TRANSFER PC12 CELLS tunneling nanotubes
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葛根素对酒精诱导的PC12氧化损伤的保护作用 预览
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作者 鲁洁 王晓丹 《泰山医学院学报》 CAS 2019年第2期92-94,共3页
目的研究葛根素(Pueraria,Pue)对酒精损伤大鼠肾上腺嗜铬细胞瘤细胞(PC12)的保护作用及其机制。方法用酒精和不同浓度葛根素处理PC12细胞;MTT法检测细胞存活率,乳酸脱氢酶(LDH)法检测葛根素对酒精致PC12细胞损伤的保护作用;相关试剂盒... 目的研究葛根素(Pueraria,Pue)对酒精损伤大鼠肾上腺嗜铬细胞瘤细胞(PC12)的保护作用及其机制。方法用酒精和不同浓度葛根素处理PC12细胞;MTT法检测细胞存活率,乳酸脱氢酶(LDH)法检测葛根素对酒精致PC12细胞损伤的保护作用;相关试剂盒测定总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化氢酶(GSH-Px)的活性和丙二醛(MDA)的量。结果葛根素能够抑制酒精诱导的PC12细胞的损伤和死亡,显著降低酒精诱导的PC12细胞的凋亡率(p<0.05),提高细胞的T-SOD、CAT和GSH-Px酶活性(p<0.05),降低MDA(p<0.05)。结论葛根素能有效保护酒精损伤的PC12细胞,其作用机理与提高细胞抗氧化能力有关。 展开更多
关键词 葛根素 酒精 PC12细胞 抗氧化 乳酸脱氢酶
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mTOR/p70S6K通路在缺氧/复氧诱导的分化后PC12神经细胞凋亡中的作用
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作者 周高雅 柴志勇 胡治平 《中南药学》 CAS 2019年第7期1024-1029,共6页
目的探讨缺氧/复氧(H/R)诱导的分化后肾上腺嗜铬细胞瘤细胞(PC12)凋亡与哺乳动物雷帕霉素靶蛋白(mTOR)/核糖体蛋白S6激酶(p70S6K)通路的关系。方法实验分为正常对照(Con)组、H/R组、H/R+N-乙酰半胱氨酸(NAC)组、H/R+雷帕霉素(Rapa)组、... 目的探讨缺氧/复氧(H/R)诱导的分化后肾上腺嗜铬细胞瘤细胞(PC12)凋亡与哺乳动物雷帕霉素靶蛋白(mTOR)/核糖体蛋白S6激酶(p70S6K)通路的关系。方法实验分为正常对照(Con)组、H/R组、H/R+N-乙酰半胱氨酸(NAC)组、H/R+雷帕霉素(Rapa)组、NAC组及Rapa组。应用MTT法检测细胞活性,Hoechst 33342染色检测细胞凋亡率,用活性氧试剂盒检测细胞内活性氧(ROS)生成,Western blot检测Bax、Bcl-2、p-mTOR(Ser2448)及p-p70S6K(Thr389)的表达。结果H/R诱导分化后PC12神经细胞凋亡,Bax表达增加,Bcl-2表达降低,NAC或Rapa显著抑制H/R所致的细胞凋亡,逆转Bax及Bcl-2表达的变化;H/R增加分化后PC12神经细胞内ROS水平,NAC显著抑制H/R所致细胞内ROS水平的增加,而Rapa对细胞内ROS水平无明显影响;H/R增加分化后PC12神经细胞p-mTOR(Ser2448)及p-p70S6K(Thr389)的表达,NAC或Rapa显著抑制H/R所致细胞p-mTOR(Ser2448)及p-p70S6K(Thr389)表达的增加。结论H/R可能通过升高细胞内ROS水平,进而激活mTOR/p70S6K信号通路,诱导分化后PC12神经细胞凋亡。 展开更多
关键词 缺氧/复氧 PC12细胞 mTOR/p70S6K通路 N-乙酰半胱氨酸 雷帕霉素
抑制PI3K/AKt信号通路对帕金森病相关基因Nurr1表达的影响 预览
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作者 庞霖霖 欧阳强 +6 位作者 韦宁 钟才 刘亚媛 周少旦 吕庚珉 何荣新 闫灵婉 《中国老年学杂志》 CAS 北大核心 2019年第15期3764-3767,共4页
目的探讨LY294002(LY)抑制磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKt)信号通路对帕金森病(PD)相关基因Nurr1表达的影响。方法体外培养BV2小胶质细胞,分为正常对照组、脂多糖(LPS)组、LY+LPS组及LY组。Western印迹检测各组PI3K/AKt信号通... 目的探讨LY294002(LY)抑制磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKt)信号通路对帕金森病(PD)相关基因Nurr1表达的影响。方法体外培养BV2小胶质细胞,分为正常对照组、脂多糖(LPS)组、LY+LPS组及LY组。Western印迹检测各组PI3K/AKt信号通路标志物P-AKt蛋白表达情况;LY抑制PI3K/AKt信号通路后,运用荧光定量(Q)-PCR检测抗炎因子白细胞介素(IL)-4、转化生长因子(TGF)-β的mRNA及PD相关基因Nurr1的表达情况,CCK-8法检测各组细胞培养液对PC12细胞存活率的影响。结果与正常对照组相比,LPS组P-AKt蛋白表达显著下降(P<0.01);与LPS组相比,LY+LPS组下降更明显(P<0.01)。与对照组相比,LPS处组IL-4、TGF-β及Nurr1mRNA表达均显著升高(均P<0.01);与LPS组相比,LY+LPS组均显著升高(均P<0.01)。与正常对照组相比,LPS培养液培养的PC12细胞存活率显著下降(P<0.01),LY预处理可以抑制LPS导致的PC12细胞死亡率。结论LY抑制PI3K/AKt信号通路能够促进BV2中PD相关基因Nurr1的表达,且对LPS诱导的炎症反应有一定的抗炎作用。 展开更多
关键词 PI3K/AKT信号通路 帕金森病 NURR1 炎症 PC12
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溶血磷脂酸诱导PC12细胞凋亡的机制研究
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作者 张杰 李易易 张兆辉 《卒中与神经疾病》 2019年第3期255-260,共6页
目的探究溶血磷脂酸诱导PC12细胞凋亡的机制并提出干预措施。方法实验1:不同浓度LPA(0μM、10μM、20μM、40μM)处理PC12细胞24 h;实验2:LPA(40μM)分别处理PC12细胞不同时间(0 h、6 h、12 h、24 h);实验3:正常组(0μM LPA)、LPA组(40... 目的探究溶血磷脂酸诱导PC12细胞凋亡的机制并提出干预措施。方法实验1:不同浓度LPA(0μM、10μM、20μM、40μM)处理PC12细胞24 h;实验2:LPA(40μM)分别处理PC12细胞不同时间(0 h、6 h、12 h、24 h);实验3:正常组(0μM LPA)、LPA组(40μM LPA)和干预组(5μM SP600125预处理2 h+40μM LPA)培养24 h;CCK-8检测细胞活力;TUNEL染色检测细胞凋亡比例;WesternBlot检测Bcl2、caspase 3、磷酸化JNK水平。结果 LPA以时间依赖和浓度依赖的方式使PC12细胞的细胞活力和Bcl2水平降低,而使PC12细胞的凋亡指数和caspase 3水平增高;SP600125(5μM)预处理不仅明显阻断LPA诱导的PC12细胞活力下降、细胞凋亡,并且极大地抑制了LPA诱导的JNK通路的激活、Bcl2水平的下调和caspase 3水平的上调。结论 JNK特异性抑制剂SP600125预处理能够明显阻断LPA诱导的PC12细胞损伤。 展开更多
关键词 溶血磷脂酸 PC12 JNK 凋亡
红景天苷对CoCl2诱导PC12细胞低氧损伤的神经保护作用 预览
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作者 杨泽霖 洪桂祝 +8 位作者 张小琴 南丽红 黄鑫 林昱 唐宇恒 刘俊杰 汪旭雯 赖文芳 褚克丹 《中国药理学通报》 CAS CSCD 北大核心 2019年第4期530-534,共5页
目的 研究红景天苷对CoCl2诱导PC12细胞低氧损伤的神经保护作用,进一步探讨红景天苷抑制补体成分,以及对神经保护的作用机制。方法 CoCl2诱导PC12细胞低氧损伤模型,经不同浓度红景天苷作用后,利用caspase-3活性检测试剂盒检测caspase-3... 目的 研究红景天苷对CoCl2诱导PC12细胞低氧损伤的神经保护作用,进一步探讨红景天苷抑制补体成分,以及对神经保护的作用机制。方法 CoCl2诱导PC12细胞低氧损伤模型,经不同浓度红景天苷作用后,利用caspase-3活性检测试剂盒检测caspase-3活性;qPCR检测Egr1、Egr4、C3等mRNA表达;Western blot检测Egr1、Egr4、C3等蛋白表达。PC12细胞低氧损伤后,C3a受体拮抗剂作用并同时给药,检测caspase-3活性、Egr1、Egr4的表达水平;siRNA干扰Egr4表达后,检测caspase-3活性及Bax、Bcl-xl、C3的表达水平。结果 红景天苷能明显降低CoCl2诱导PC12细胞低氧损伤模型的caspase-3活性及Bax、C3的蛋白表达,促进Bcl-xl、Egr1、Egr4的蛋白表达;C3a受体拮抗剂作用后,能够抑制caspase-3活性及Egr1、Egr4的表达;siRNA干扰Egr4表达后,能逆转caspase-3活性及Bax、Bcl-xl的表达水平,但对C3作用不明显。结论 红景天苷对CoCl2诱导PC12细胞低氧损伤模型具有神经保护作用,与红景天苷抑制补体成分C3,促进Egr4的表达水平,进而抑制细胞凋亡有关。 展开更多
关键词 红景天苷 低氧损伤 PC12 补体 神经保护 Egr4
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白藜芦醇通过激活mTOR/自噬抑制缺氧缺糖/复氧复糖诱导的PC12细胞损伤及凋亡 预览 被引量:1
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作者 吴印华 胡万保 《中医药临床杂志》 2019年第2期298-303,共6页
目的:白藜芦醇对缺氧缺糖/复氧复糖(OGD/R)诱导的PC12细胞损伤及凋亡的影响及作用机制。方法:以PC12细胞建立OGD/R自噬性损伤模型,设置空白对照组、模型组、不同浓度(5、10μmol/L)白藜芦醇组,自噬抑制剂(3-MA)组。使用CCK-8试剂盒检测P... 目的:白藜芦醇对缺氧缺糖/复氧复糖(OGD/R)诱导的PC12细胞损伤及凋亡的影响及作用机制。方法:以PC12细胞建立OGD/R自噬性损伤模型,设置空白对照组、模型组、不同浓度(5、10μmol/L)白藜芦醇组,自噬抑制剂(3-MA)组。使用CCK-8试剂盒检测PC12的增殖能力;流式细胞术检测各组细胞凋亡率;ELISA检测各组细胞培养液中TNF-α和IL-6的浓度变化;DHE荧光探针检测ROS的变化;特定试剂盒检测乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH-PX)含量;Westernblot法检测各组mTOR的磷酸化情况及LC3-Ⅱ、Beclin-1蛋白表达。结果:与空白对照组相比模型组能够显著降低PC12的增殖能力,增加细胞中ROS的含量和凋亡率,诱导TNF-α和IL-6分泌,LDH、MDA含量显著增加,SOD、GSH-PX活性显著下降,而P-mTOR/mTOR、LC3-Ⅱ、Beclin-1表达增加(P<0.01)。与模型组比较,不同浓度白藜芦醇作用PC12时,细胞存活率逐渐上升,ROS含量及凋亡率逐渐下降,LDH、MDA含量减少,SOD、GSH-PX活性随之上升,TNF-α和IL-6分泌显著下降,P-mTOR/mTOR、LC3-Ⅱ、Beclin-1表达进一步增加(P<0.01)。自噬抑制剂(3-MA)组上述指标的检测结果与白藜芦醇组相反。结论:白藜芦醇可能部分通过激活mTOR/自噬通路对神经细胞发挥保护作用。 展开更多
关键词 白藜芦醇 PC12 凋亡 mTOR/自噬
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原儿茶酸对碱性成纤维细胞生长因子诱导PC12细胞分化的影响
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作者 王丽芳 薛小燕 谢宁生 《抗感染药学》 2019年第6期931-933,1008共4页
目的:探究原儿茶酸(PCA)对碱性成纤维细胞生长因子(bFGF)诱导PC12细胞分化的影响。方法:体外培养PC12细胞,采用PCA低、中和高剂量组(16.0、32.0和64.0μmol/L)增强bFGF诱导PC12细胞分化,并通过其细胞形态学的观察、细胞活力的检测,分析... 目的:探究原儿茶酸(PCA)对碱性成纤维细胞生长因子(bFGF)诱导PC12细胞分化的影响。方法:体外培养PC12细胞,采用PCA低、中和高剂量组(16.0、32.0和64.0μmol/L)增强bFGF诱导PC12细胞分化,并通过其细胞形态学的观察、细胞活力的检测,分析突起细胞数目及平均突起长度值,考察不同剂量的PCA对bFGF诱导PC12细胞分化的影响。结果:与对照组相比,低、中和高剂量(16.0、32.0和64.0μmol/L)PCA组能增强bFGF诱导PC12细胞分化。结论:PCA能增强bFGF诱导PC12细胞分化的能力。 展开更多
关键词 原儿茶酸(PCA) 碱性成纤维细胞生长因子(bFGF) PC12 细胞分化
多巴胺抑制Erastin诱导PC12细胞系铁死亡 预览
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作者 高万里 王鼎 +2 位作者 李晓莹 陈永源 周伯荣 《基础医学与临床》 CSCD 2019年第2期207-212,共6页
目的探讨多巴胺在不同诱导剂导致的嗜铬细胞瘤PC12细胞死亡中的作用。方法应用RT-qPCR鉴定PC12的肿瘤特异性基因和神经相关基因的mRNA表达;通过细胞存活率评价生理浓度范围内(0~100μmol/L)不同浓度(0、6.25、12.5、25、50和100μmol/L... 目的探讨多巴胺在不同诱导剂导致的嗜铬细胞瘤PC12细胞死亡中的作用。方法应用RT-qPCR鉴定PC12的肿瘤特异性基因和神经相关基因的mRNA表达;通过细胞存活率评价生理浓度范围内(0~100μmol/L)不同浓度(0、6.25、12.5、25、50和100μmol/L)的多巴胺对PC12存活的影响,以及对药物诱导的铁死亡、凋亡和坏死的影响;流式细胞仪检测并分析PC12细胞铁死亡的发生和多巴胺保护PC12过程中PI、Annexin V和细胞周期的情况。结果PC12表达肿瘤特异性基因和部分神经类基因。多巴胺在浓度25、50和100μmol/L时对PC12细胞有杀伤效果。生理浓度多巴胺特异性保护Erastin诱导的PC12细胞铁死亡。多巴胺能有效的降低Erastin造成的PI、Annexin V双阳性细胞上升和S期到G2/M期阻滞。结论生理浓度多巴胺对PC12有细胞毒性,并可以保护PC12抑制Erastin诱导的铁死亡。 展开更多
关键词 PC12 多巴胺 细胞死亡 铁死亡 Erastin
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Abnormal mitochondrial fusion and fissionin PBDE-47-induced change in mitochondrial mass in PC12 cells
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作者 杨凯朝 《中国医学文摘:内科学分册(英文版)》 2019年第2期73-74,共2页
Objective To investigate the effect of 2,2’,4,4’-tetrabromodiphenyl ether (PBDE-47) on the mitochondrial mass in rat adrenal pheochromocytoma (PC12)cells and the potential mechanisms. Methods Highly differentiated P... Objective To investigate the effect of 2,2’,4,4’-tetrabromodiphenyl ether (PBDE-47) on the mitochondrial mass in rat adrenal pheochromocytoma (PC12)cells and the potential mechanisms. Methods Highly differentiated PC12 cells were divided into control,1,10 or20μmol/L PBDE-47-treated groups and cultured for 24 h. 展开更多
关键词 PC ABNORMAL MITOCHONDRIAL FUSION PC12 CELLS
Ginsenoside Rgl protects against ischemic/reperfusioninduced neuronal injury through miR-144/Nrf2/ARE pathway
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作者 Shi-feng Chu Zhao Zhang +10 位作者 Xin Zhou Wen-bin He Chen Chen Piao Luo Dan-dan Liu Qi-di Ai Hai-fan Gong Zhen-zhen Wang Hong-shuo Sun Zhong-ping Feng Nai-hong Chen 《中国药理学报:英文版》 SCIE CAS CSCD 2019年第1期13-25,共13页
Ginsenoside Rg1 (Rg1),a saponin extracted from Panax ginseng,has been well documented to be effective against ischemic/ reperfusion (I/R)neuronal injury.However,the underlying mechanisms remain obscure.In the present ... Ginsenoside Rg1 (Rg1),a saponin extracted from Panax ginseng,has been well documented to be effective against ischemic/ reperfusion (I/R)neuronal injury.However,the underlying mechanisms remain obscure.In the present study,we investigated the roles of Nrf2 and miR-144 in the protective effects of Rgl against I/R-induced neuronal injury.In OGD/R-treated PC12 cells,Rgl (0.01-1 μmol/L)dose-dependently attenuated the cell injury accompanied by prolonging nuclear accumulation of Nrf2,enhancing the transcriptional activity of Nrf2,as well as promoting the expression of ARE-target genes.The activation of the Nrf2/ARE pathway by Rgl was independent of disassociation with Keapl,but resulted from post-translational regulations.Knockdown of Nrf2 abolished all the protective changes of Rgl in OG-D/R-treated PC12 cells.Furthermore,Rgl treatment significantly decreased the expression of miR-144,which downregulated Nrf2 production by targeting its 3'-untranlated region after OGD/R.Knockdown of Nrf2 had no effect on the expression of miR-144,suggesting that miR-144 was an upstream regulator of Nrf2.We revealed that there was a direct binding between Nrf2 and miR-144 in PC12 cells.Application of anti-miR-144 occluded the activation of the Nrf2/ ARE pathway by Rgl in OGD/R-treated PC12 cells.In tMCAO rats,administration of Rgl (20 mg/kg).significantly alleviated ischemic injury,and activated Nff2/ARE pathway.The protective effects of Rgl were abolished by injecting of AAV-HIF-miR-144-shRNA into the predicted ischemic penumbra.In conclusion,our results demonstrate that Rgl alleviates oxidative,stress after I/R through inhibiting miR-144 activity and subsequently promoting the Nrf2/ARE pathway at the post-translational-level. 展开更多
关键词 STROKE GINSENOSIDE Rgl ischemic/reperfusion oxidative stress Nrf2/ARE miR-144 PC12 cells tMCAO rats
Mu-Xiang-You-Fang protects PC12 cells against OGD/R-induced autophagy via AMPK/mTOR signaling pathway 预览
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作者 MA Hui-xia CHEN Ai-ling +3 位作者 HOU Fan LI Ting-ting ZHU Ya-fei ZHAO Qi-peng 《中国药理学与毒理学杂志》 CAS 北大核心 2019年第9期742-743,共2页
OBJECTIVE Mu-Xiang-You-Fang(MXYF)is a classic prescription of Hui medicine,composed of five herbs,which has been used to treat ischemic stroke for many years.However,the potential pharmacological mecha⁃nisms of MXYF r... OBJECTIVE Mu-Xiang-You-Fang(MXYF)is a classic prescription of Hui medicine,composed of five herbs,which has been used to treat ischemic stroke for many years.However,the potential pharmacological mecha⁃nisms of MXYF remain unclear.The present research is to investigate the neuroprotective effect of MXYF and its role in modulating autophagy via AMPK/mTOR signaling pathway in the PC12 oxygen-glucose deprivation and reperfusion(OGD/R)injury model.METHODS MXYF was extracted by supercritical CO2 fluid extraction apparatus.PC12 OGD/R injury model was established by oxygen-glucose deprivation for 2 h and reperfusion for 24 h.The effects of MXYF on the viability and cytotoxicity of PC12 cells were determined through cell counting kit(CCK-8)assay.Colorimetric method was performed to determine the LDH leakage rate.The calcium concentration was determined by chemical fluorescence method and the mitochondrial membrane potential was determined through flow cytometry.Monodansylcadaverine(MDC)staining was conducted to detect autophagosome formation.The expression of LC3,Beclin1,p62,p-AMPK,ULK1,p-mTOR and p-p70s6k proteins were determined by immunofluorescence and Western blotting analyses.RESULTS MXYF(1,2 and 4 mg·L^-1)could significantly increase the cell viability and mitochondrial membrane potential,while decreased the release of lactate dehydrogenase(LDH)and calcium concentration in PC12 cells.Mechanistic studies showed that MXYF reduced the LC3-II/LC3-I ratio and inhibited the expression of beclin1,p-AMPK and ULK1.In comparison,the expres⁃sion of p-mTOR,p-p70s6k and p62 were significantly enhanced.CONCLUSION MXYF inhibits autophagy after OGD/Rinduced PC12 cell injury through AMPK-mTOR pathway,thus MXYF might have therapeutic potential for treating the ischemic stroke. 展开更多
关键词 Mu-Xiang-You-Fang PC12 cells oxygen-glucose deprivation and reperfusion AUTOPHAGY AMPK/mTOR pathway
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Long non-coding RNA GAS5 promotes PC12 cells differentiation into Tuj1-positive neuron-like cells and induces cell cycle arrest 预览
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作者 He-Yan Zhao Sheng-Tong Zhang +7 位作者 Xiang Cheng Hao-Ming Li Lei Zhang Hui He Jian-Bing Qin Wei-Ye Zhang Yan Sun Guo-Hua Jin 《中国神经再生研究:英文版》 SCIE CAS CSCD 2019年第12期2118-2125,共8页
Growth arrest-specific 5 (GAS5) is an anti-oncogene that has been extensively studied in tumors. However, research on GAS5 in the context of nervous system disease is rare at present. This study aimed to investigate t... Growth arrest-specific 5 (GAS5) is an anti-oncogene that has been extensively studied in tumors. However, research on GAS5 in the context of nervous system disease is rare at present. This study aimed to investigate the role of the long non-coding RNA GAS5 in rat pheochromocytoma cells (PC12 cells). GAS5-overexpressing lentivirus was transfected into PC12 cells, and expression levels of GAS5 and C-myc were detected by real-time PCR. Ratios of cells in S phase were detected by 5-ethynyl-2′-deoxyuridine. Immunohistochemical staining was used to detect the immunoreactivity of neuron microtubule markers Tuj1, doublecortin, and microtubule-associated protein 2. Apoptosis was detected by flow cytometry, while expression of acetylcholine in cells was detected by western blot assay. We found that GAS5 can promote PC12 cells to differentiate into Tuj1-positive neuron-like cells with longer processes. In addition, cell proliferation and cell cycle were significantly suppressed by GAS5, whereas it had no effect on apoptosis of PC12 cells. Our results indicate that GAS5 could increase the expression of choline acetyltransferase and acetylcholine release. Thus, we speculate that GAS5 is beneficial to the recovery of neurons and the cholinergic nervous system. 展开更多
关键词 nerve REGENERATION growth arrest-specific 5 PC12 CELL neuron proliferation CELL cycle CHOLINE ACETYLTRANSFERASE ACETYLCHOLINE Alzheimer's disease neural REGENERATION
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葡萄籽提取物对PC12神经元毒性的研究 预览
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作者 杨智超 宋丽娟 +2 位作者 姜维佳 宋俊丽 王青 《山西中医学院学报》 2018年第2期13-15,共3页
目的:考察葡萄籽提取物(grape seed extract,GSE)在体外对高分化的PC12神经元增殖的影响及机制。方法:采用MTT法考察GSE对PC12神经元增殖的影响,Western blot法检测GSE对PC12神经元中与凋亡相关的蛋白Bcl-2、Bax表达量的影响。结果... 目的:考察葡萄籽提取物(grape seed extract,GSE)在体外对高分化的PC12神经元增殖的影响及机制。方法:采用MTT法考察GSE对PC12神经元增殖的影响,Western blot法检测GSE对PC12神经元中与凋亡相关的蛋白Bcl-2、Bax表达量的影响。结果:实验结果表明,低浓度的GSE(2.5μg/m L,5μg/m L)长时间作用72 h可促进PC12细胞增殖,10μg/m L GSE对PC12细胞增殖无影响,20μg/m L GSE作用48 h出现细胞毒性,30μg/m L,50μg/m L和100μg/m L GSE作用24 h即可抑制细胞增殖。GSE显著抑制抗凋亡蛋白Bcl-2的表达,对促凋亡蛋白Bax表达无影响。结论:20μg/m L以上浓度的GSE作用于PC12神经元会显著抑制其增殖,作用机制为减少了细胞抗凋亡蛋白Bcl-2的表达。 展开更多
关键词 葡萄籽提取物 PC12 细胞毒性
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去氢骆驼蓬对PC12和NCTC1469细胞增殖与凋亡的影响 被引量:1
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作者 陈蓓 马丽娟 +4 位作者 李丽华 文丽梅 高惠静 王建华 赵军 《中国病原生物学杂志》 CSCD 北大核心 2018年第1期59-63,67共6页
目的 探讨去氢骆驼蓬碱Harmine(HM)对小鼠肝细胞NCTC1469和神经细胞PC12增殖与凋亡的影响,为去氢骆驼蓬碱衍生物作为抗包虫病候选药物的研发提供参考。方法 体外培养的小鼠肝细胞NCTC1469和神经细胞PC12分为HM干预组和无药物组对照组... 目的 探讨去氢骆驼蓬碱Harmine(HM)对小鼠肝细胞NCTC1469和神经细胞PC12增殖与凋亡的影响,为去氢骆驼蓬碱衍生物作为抗包虫病候选药物的研发提供参考。方法 体外培养的小鼠肝细胞NCTC1469和神经细胞PC12分为HM干预组和无药物组对照组,用噻唑蓝(MTT)比色法测定不同浓度HM作用不同时间对细胞增殖的抑制作用;倒置显微镜下观察药物作用后的细胞形态改变;采用Hoechst 33258核染色法观察细胞凋亡的形态学变化,采用流式细胞术检测细胞凋亡情况。结果 MTT检测去氢骆驼蓬碱可抑制PC12和NCTC1469和细胞的增殖,该抑制作用呈时间-浓度依赖(F值分别为126.43、125.32、145.82,P〈0.01;F值分别为147.95、222.67、242.28,P〈0.01)。HM处理PC12细胞24、48、72h的IC50分别为32.40、24.50和15.37μg/ml,HM处理PC12细胞24、48、72h对NCTC1469细胞的IC50分别为62.19、38.43和28.52μg/ml。对照组细胞贴壁生长,排列均匀,大小基本一致,轮廓清晰,悬浮细胞少;实验组随着去氢骆驼蓬碱浓度的升高,细胞出现皱缩,细胞数量及生长密度逐渐减少。Hoeehst33258染色观察去氢骆驼蓬碱干预24h后细胞核均出现不同程度蓝色荧光,可见核致密、浓染等典型的细胞凋亡特征。10、20、40μg/ml去氢骆驼蓬碱干预24h,PC12细胞总凋亡率分别为(12.52±2.21)%、(23.44±1.27)和(55.85±2.30)%,NCTC1469细胞总凋亡率分别为(15.20±2.67)%、(41.31±3.09)和(53.92±2.58)%,与对照组比较差异均有统计学意义(t值分别为-27.42、-48.73、-93.76,P〈0.01;t值分别为-76.00、-116.67、-144.25,P〈0.01)。结论 去氢骆驼蓬碱能抑制PC12、NCTC1469细胞增殖,促进细胞凋亡,且存在正向的剂量-效应关系。 展开更多
关键词 去氢骆驼蓬碱 增殖 凋亡 PC12细胞 NCTC1469细胞
梓醇调控自噬和凋亡促进H2O2诱导的PC12细胞存活 被引量:2
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作者 王园 刘珂 +3 位作者 邱邴勋 祝慧凤 王涛 万东 《西南大学学报:自然科学版》 CSCD 北大核心 2018年第2期27-34,共8页
探讨梓醇对过氧化氢(H2O2)诱导的PC12细胞氧化损伤后调控凋亡和自噬相关分子的机制,采用0.01 mmol,0.1 mmol,1 mmol的梓醇预处理PC12细胞2h后,再用250μmol/L H2O2损伤细胞12h.MTT法检测细胞存活率,设置梓醇低、中、高3个剂量组(0.01... 探讨梓醇对过氧化氢(H2O2)诱导的PC12细胞氧化损伤后调控凋亡和自噬相关分子的机制,采用0.01 mmol,0.1 mmol,1 mmol的梓醇预处理PC12细胞2h后,再用250μmol/L H2O2损伤细胞12h.MTT法检测细胞存活率,设置梓醇低、中、高3个剂量组(0.01 mmol,0.1 mmol,1 mmol),自噬抑制剂3MA阳性对照组,阴性对照组,H2O2诱导的PC12细胞损伤模型组,采用荧光单标检测LC3Ⅱ的表达,Western blot检测凋亡和自噬相关蛋白Bcl2/Bax,LC3Ⅱ/Ⅰ和Cleaved-caspase 3的表达情况.结果显示,梓醇呈剂量依赖性地提高了H2O2诱导的PC12细胞的存活,并对H2O2诱导的PC12细胞的LC3Ⅱ/Ⅰ,Bcl2/Bax升高蛋白水平表达,降低了Cleaved-caspase 3的表达;随着梓醇浓度的增加,Cleaved-caspase 3的表达减少,而LC3Ⅱ/Ⅰ和Bcl2/Bax的表达在升高.得出梓醇通过恢复凋亡和自噬相关蛋白Bcl2/Bax,LC3Ⅱ/Ⅰ的平衡能够保护PC12细胞免于H2O2诱导的氧化损伤. 展开更多
关键词 梓醇 PC12细胞 过氧化氢 凋亡 自噬
基于析因设计的川芎嗪和阿魏酸配伍抗PC12氧糖剥夺损伤的交互作用研究 被引量:1
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作者 刘璐 徐士欣 +2 位作者 张军平 张男男 杨惠林 《药物评价研究》 CAS 2018年第9期1597-1601,共5页
目的观察川芎嗪和阿魏酸配伍对PC12糖氧剥夺(OGD)损伤的预保护作用,优化配比组合。方法体外培养PC12,考虑A(是否OGD造模)、B(川芎嗪0.5、1.0、2.0μmol/L)及C(阿魏酸5、10、20μmol/L)3个因素,采用完全随机分组的2×3×... 目的观察川芎嗪和阿魏酸配伍对PC12糖氧剥夺(OGD)损伤的预保护作用,优化配比组合。方法体外培养PC12,考虑A(是否OGD造模)、B(川芎嗪0.5、1.0、2.0μmol/L)及C(阿魏酸5、10、20μmol/L)3个因素,采用完全随机分组的2×3×3析因设计。倒置相差显微镜观察造模前后细胞形态;MTS测细胞活力;采用乳酸脱氢酶(LDH)法测定细胞损伤,并进行析因分析。结果与对照组比较,模型组细胞变圆或肿胀,突起回缩或消失,细胞膜褶皱或破损,胞体折光性差,细胞数目明显减少,细胞活力降低;与模型组比较,加药组细胞损伤得到不同程度的改善,细胞活力增加,川芎嗪2μmol/L+阿魏酸20μmol/L改善作用最明显。A、B、C 3因素均对LDH值具有显著影响(P〈0.01),且B、C间存在交互作用(P〈0.01),以川芎嗪2μmol/L、阿魏酸20μmol/L配比时,LDH值最低。结论川芎嗪与阿魏酸合用对PC12剥夺糖氧损伤具有预保护作用,川芎嗪与阿魏酸配比具有交互作用,最佳配比为2∶20。 展开更多
关键词 川芎嗪 阿魏酸 PC12细胞 析因分析
五子衍宗丸细胞干预液的制备及其抗NTDs细胞凋亡的作用研究
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作者 孙琳 姜弢 +3 位作者 樊慧杰 张娜 严维花 柴智 《山西大学学报:自然科学版》 CSCD 北大核心 2018年第3期650-655,共6页
采用超高效液相色谱法对五子衍宗丸中金丝桃苷的含量进行了测定,并比较了70℃蒸馏水、常温蒸馏水、体积分数70%乙醇、无水乙醇和DMSO等5种溶剂对金丝桃苷的提取效果;在此基础上,优选出细胞干预液制备方法,观察五子衍宗丸对NTDs细胞凋亡... 采用超高效液相色谱法对五子衍宗丸中金丝桃苷的含量进行了测定,并比较了70℃蒸馏水、常温蒸馏水、体积分数70%乙醇、无水乙醇和DMSO等5种溶剂对金丝桃苷的提取效果;在此基础上,优选出细胞干预液制备方法,观察五子衍宗丸对NTDs细胞凋亡的防控作用。结果显示,金丝桃苷在0.004~0.1μg范围内具有良好的线性关系(r=0.999 9);平均回收率为98.85%,RSD为3.86%;5种提取溶剂中体积分数为70%乙醇对金丝桃苷的提取效果最好;以五子衍宗丸70%乙醇提取物干预NTDs细胞,能够明显提高细胞存活率,有效抑制细胞凋亡。 展开更多
关键词 金丝桃苷 五子衍宗丸 超高效液相色谱法 神经管畸形 PC12细胞
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