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Science China Chemistry 2015年第10期58卷 1600-1604页

作者： Pratikkumar Shah Qiaoli Yue Xuena Zhu Fangcheng Xu Huai-Sheng Wang Chen-Zhong Li College of Chemistry and Chemical Engineering Liaocheng University Nanobioengineering/Bioelectronics Laboratory Department of Biomedical Engineering Florida International University Department of Chemical and Biochemical Engineering College of Chemical Engineering Xiamen University

In this research, we explored a rapid assessment of silver nanoparticles(Ag-NPs) neurotoxicity at a single-cell level. Traditional nanotoxicity assays on large cell-populations may hide the important heterogeneity of individual cells often found in neuronal cells. The development in the area of new nanomaterial discoveries is far ahead of the development of advanced tools to measure these materials' toxicity. Development of alternative approaches to assess nanomaterials toxicity rapidly, reliably, and accurately is desirable. Here, we present a chip-based, cell-integrated microwell-array device for rapid assessment of neurotoxicity of Ag-NPs by monitoring the exocytosis function of a pc12 cell. Results presented here confirm the dose-dependent toxicity of Ag-NPs and the immediate alteration of their exocytosis function when exposed to NPs.

关键词： neuron devices nanotoxicity amperometry exocytosis pc12 silver nanoparticle

Ligustilide protects pc12 cells from oxygen-glucose deprivation/reoxygenation-induced apoptosis via the LKB1-AMPK-mTOR signaling pathway

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Neural Regeneration Research 2020年第3期15卷 473-481页

作者： Dan-Yang Zhao Dong-Dong Yu Li Ren Guo-Rong Bi Shengjing Hospital of China Medical University ShenyangLiaoning ProvinceChina The First People’s Hospital of Shenyang ShenyangLiaoning ProvinceChina The First Affiliated Hospital of Liaoning University of Traditional Chinese Medicine ShenyangLiaoning ProvinceChina

Autophagy has been shown to have a protective effect against brain ***(LIG)is a bioactive substance isolated from Ligusticum chuanxiong,a traditional Chinese *** has a neuroprotective effect;however,it is unclear whether this neuroprotective effect involves *** 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)*** with 1×10^-6 M LIG for 3 hours had the greatest effect on cell proliferation,and was therefore used for subsequent ***12 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 *** was detected by flow *** 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 *** staining was used to detect LC3-II *** formation was observed by electron *** 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/*** 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 *** together,our findings show that LIG promotes autophagy and protects

关键词： AMPK apoptosis autophagy Bax Bcl-2 Beclin 1 LC3-II ligustilide mTOR pc12 cells

Garcinia xanthochymus extract protects pc12 cells from H2O2-induced apoptosis through modulation of PI3K/AKT and NRF2/HO-1 pathways

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Chinese Journal of Natural Medicines 2017年第11期15卷 825-833页

作者： XU Jing GAN Sheng LI Jun WAND De-Bing CHEN Yu HU Xin YANG Guang-Zhong Laboratory for Natural Products Chemistry School of Pharmaceutical Sciences South-Central University for NationalitiesWuhan 430074 China College of Chemistry and Material Sciences South Central University for Nationalities Wuhan 430074 China

The aim of the present study was to investigate the protective effects and underlying mechanisms of Garcinia xanthochymus, a perennial medicinal plant native to Yunnan, China, against H2 O2-induced oxidative damage in rat pheochromacytoma pc12 cells. Preincubation of pc12 cells with fruit Et OAc fraction(fruit-EFr., 12.5–50 μmol·L^(-1)) of G. xanthochymus for 24 h prior to H_2O_2 exposure markedly improved cell viability and increased the activities of antioxidant enzymes(superoxide dismutase, catalase, and heme oxygenase-1 [HO-1]), prevented lactate dehydrogenase release and lipid peroxidation malondialdehyde production, attenuated the decrease of matrix metalloproteinases(MMP), and scavenged reactive oxygen species(ROS). Fruit-EFr. also reduced BAX and cytochrome C expression and improved BCL-2 expression, thereby decreasing the ratio of BAX to BCL-2. Fruit-EFr. activated the nuclear translocation of NRF2 to increase HO-1 and induced the phosphorylation of AKT. Its cytoprotective effect was abolished by LY294002, a specific inhibitor of PI3 K. Taken together, the above findings suggested that *** G. xanthochymus could enhance cellular antioxidant defense capacity, at least in part, through upregulating HO-1 expression and activating the PI3 K/AKT pathway and that it could suppress H_2O_2-induced oxidative damage via PI3 K/AKT and NRF2/HO-1 signaling pathways.

关键词： Garcinia xanthochymus Oxidative stress pc12 PI3K/Akt pathway Nrf2/HO-1 signaling pathways

Ganoderic Acids A and B Reduce Okadaic Acid-Induced Neurotoxicity in pc12 Cells by Inhibiting Tau Hyperphosphorylation

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Biomedical and Environmental Sciences 2023年第1期36卷 103-108页

作者： CUI Jing MENG Yu Han WANG Zhan Wei WANG Jing SHI Dong Fang LIU Duo Central Laboratory Changchun Normal UniversityChangchun 130032JilinChina School of Life Sciences Northeast Normal UniversityChangchun 130024JilinChina

Alzheimer’s disease is one of the most debilitating neurodegenerative diseases affecting the aging population. One of the neuropathological hallmarks of AD is an aggregation of the tau protein into neurofibrillary tangles(NFTs) in neurons and glial cells of the brain. Tau is a microtubuleassociated protein expressed mainly in neurons of the central nervous system[1].

关键词： Alzheimer tau pc12

Long non-coding RNA GAS5 promotes pc12 cells differentiation into Tuj1-positive neuron-like cells and induces cell cycle arrest

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Neural Regeneration Research 2019年第12期14卷 2118-2125页

作者： He-Yan Zhao Sheng-Tong Zhang Xiang Cheng Hao-Ming Li Lei Zhang Hui He Jian-Bing Qin Wei-Ye Zhang Yan Sun Guo-Hua Jin Department of Human Anatomy the Jiangsu Key Laboratory of Neuroregeneration Medical School Nantong University

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.

关键词： nerve regeneration growth arrest-specific 5 pc12 cell neuron proliferation cell cycle choline acetyltransferase acetylcholine Alzheimer's disease neural regeneration

Transfer of mitochondria from mesenchymal stem cells derived from induced pluripotent stem cells attenuates hypoxia-ischemia-induced mitochondrial dysfunction in pc12 cells

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Neural Regeneration Research 2020年第3期15卷 464-472页

作者： Yan Yang Gen Ye Yue-Lin Zhang Hai-Wei He Bao-Qi Yu Yi-Mei Hong Wei You Xin Li Department of Emergency Medicine Department of Emergency and Critical Care MedicineGuangdong Provincial People’s HospitalGuangdong Academy of Medical SciencesGuangzhouGuangdong ProvinceChina Department of Emergency the First Affiliated HospitalSun Yat-sen UniversityGuangzhouGuangdong ProvinceChina Shantou University Medical College ShantouGuangdong ProvinceChina Department of Physiology and Pathophysiology School of Basic Medical SciencesCapital Medical UniversityKey Laboratory of Remodelling-related Cardiovascular DiseasesMinistry of EducationBeijingChina

Mitochondrial dysfunction in neurons has been implicated in hypoxia-ischemia-induced brain *** mesenchymal stem cell therapy has emerged as a novel treatment for this pathology,the mechanisms are not fully *** 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 *** oxygen species in pc12 cells was measured by *** membrane potential(ΔΨm)in pc12 cells was determined by JC-1 *** of pc12 cells was detected by terminal deoxynucleotidal transferase-mediated dUTP nick end-labeling *** morphology in pc12 cells was examined by transmission electron *** of mitochondria from the mesenchymal stem cells derived from induced pluripotent stem cells to damaged pc12 cells was measured by flow *** stem cells were induced from pluripotent stem cells by lentivirus infection containing green fluorescent protein in *** they were co-cultured with pc12 cells in Transwell chambers and treated with CoCl2 for 24 hours to detect adenosine triphosphate level in pc12 ***2-induced pc12 cell damage was ***-culture with mesenchymal stem cells significantly reduced apoptosis and restoredΔΨm in the injured pc12 cells under CoCl2 ***-culture with mesenchymal stem cells ameliorated mitochondrial swelling,the disappearance of cristae,and chromatin margination in the injured pc12 *** 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 *** transfer efficiency was greatly enhanced in the presence of *** importantly,inhibition of tunneling nanotubes partially abrogated the beneficial effects of mesenchymal stem cells on CoCl2-induced pc12 cell injury.M

关键词： apoptosis brain injury hypoxia-ischemia induced pluripotent stem cells mesenchymal stem cells mitochondrial membrane potential mitochondrial transfer pc12 cells tunneling nanotubes

Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in pc12 cells subjected to oxygen-glucose deprivation

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Neural Regeneration Research 2020年第11期15卷 2154-2161页

作者： Tao Pan Qiu-Jiao Zhu Li-Xiao Xu Xin Ding Jian-Qin Li Bin Sun Jun Hua Xing Feng Department of Neonatology Children’s Hospital Affiliated to Suzhou UniversitySuzhouJiangsu ProvinceChina Department of Critical Care Medicine Children’s Hospital Affiliated to Suzhou UniversitySuzhouJiangsu ProvinceChina Institute of Pediatrics Children’s Hospital Affiliated to Suzhou UniversitySuzhouJiangsu ProvinceChina Blood Section Children’s Hospital Affiliated to Suzhou UniversitySuzhouJiangsu ProvinceChina

Transient receptor potential melastatin 2(TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma(pc12) cells injured by oxygen-glucose deprivation(OGD). pc12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2(CXCL2), NACHT, LRR, and PYD domain–containing protein 3(NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.

关键词： apoptosis calcium caspase-1 NLRP3 mitochondrial impairment oxidative stress oxygen-glucose deprivation pc12 shRNA TRPM2

Protective effects of organic extracts of Alpinia oxyphylla against hydrogen peroxide-induced cvtotoxicitv in pc12 cells

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Neural Regeneration Research 2020年第4期15卷 682-689页

作者： Li-Hong Duan Meng Li 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 Shenzhen Institute of Geriatrics ShenzhenGuangdong ProvinceChina Department of Rehabilitation the Second People’s Hospital of ShenzhenShenzhenGuangdong ProvinceChina Stroke Biological Recovery Laboratory Spaulding Rehabilitation HospitalHarvard Medical SchoolCharlestownMAUSA Department of Mechanical and Biomedical Engineering City University of Hong KongHong Kong Special Administrative RegionChina Department of Rehabilitation the First Affiliated Hospital of Shenzhen UniversityShenzhenGuangdong ProvinceChina School of Mechanical and Automotive Engineering South China University of TechnologyGuangzhouGuangdong ProvinceChina School of Mechanical Engineering Guangxi University of Science and TechnologyLiuzhouGuangxi Zhuang Autonomous RegionChina Mingkai Smart Medical Robot Co. Ltd.ShenzhenGuangdong ProvinceChina Kerry Rehabilitation Medicine Research Institute ShenzhenGuangdong ProvinceChina

Alpinia oxyphylla,a traditional herb,is widely used for its neuroprotective,antioxidant and memory-improving ***,the neuroprotective mechanisms of action of its active ingredients are *** 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 *** 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 *** 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 *** ethyl acetate,petroleum ether and n-butanol extracts(each tested at 100,150 and 200μg/mL)had neuroprotective effects as *** optimum effective concentration ranged from 50–200μg/mL,and the protective effect of the ethyl acetate extract was comparatively *** results demonstrate that organic extracts of Alpinia oxyphylla protect pc12 cells against apoptosis induced by hydrogen *** findings should help identify the bioactive neuroprotective components in Alpinia oxyphylla.

关键词： active ingredients Alpinia oxyphylla apoptosis ethanol crude extract fraction hydrogen peroxide nerve regeneration neuroprotective agent neuroprotective effects pc12 cells traditional herb

Signal transduction pathway of glucocorticoid's rapid action in pc12 cells

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Chinese Science Bulletin 1999年第20期44卷 1837-1843页

作者： Jian Qiu Shujie Lou Yizhang Chen Neuroscience Research Institute Second Military Medical University Shanghai China

Traditionally, steroid hormone action has been described as the modulation of nuclear transcription, thus triggering genomic events that are responsible for physiological effects. Despite early observation of rapid steroid effects that were incompatible with this theory, nonge-nomic steroid action has been widely recognized only recently. More and more evidence for these rapid effects has been reported in recent years, but the signal transduction pathways

关键词： pc12 cells glucocorticoid nongenomic signal transduction.

Neuronal growth inhibitory factor inhibits pheochromo-cytoma pc12 in vitro

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Chinese Science Bulletin 2002年第2期47卷 132-135页

作者： Ji, QZ Ru, BG Peking Univ Natl Lab Prot Engn Coll Life Sci Beijing 100871 Peoples R China

Neuronal growth inhibitory factor (GIF), named Metallothioneins-III (MT-III), is the first protein validated to be capable of inhibiting the growth of neurons in nervous system. We have detected the effects of recombinant GIF on the growth of neuroblastoma SY5Y and pheochro-mocytoma pc12 by the MTT (Thiazolyl blue) reduction as-say. Recombinant GIF inhibited pc12 in vitro; the inhibitory rate was about 25% when GIF was at 100 mg/L; and the inhibitory rate was about 50% when GIF was at 300 mg/L. It is shown that pc12 could serve as a proper model for de-tecting neuronal growth inhibitory activity of GIF. Recom-binant GIF did not inhibit neuroblastoma SY5Y in vitro, a common model of neuroma; it is also shown that GIF could not inhibit neuromata extensively. The reason for GIF inhib-iting pc12 may be that pc12 have some properties of cho-linergic neuron. It must play an important role in discover-ing the mechanism of GIF’s neuronal growth inhibitory ac-tivity.

关键词： neuronal growth inhibitory factor pheochromocytoma pc12 neuroblastoma SY5Y MTT reduction assay.