β淀粉样蛋白25~35 对PC12细胞骨架的毒性作用

曹晶晶 范文娟 石贞玉 鄢明超 刘红亮 王来 邓锦波*

doi:10.16098/j.issn.0529-1356.2016.04.06

PDF(752 KB)

解剖学报 ›› 2016, Vol. 47 ›› Issue (4) : 469-475. DOI: 10.16098/j.issn.0529-1356.2016.04.06

细胞和分子生物学

β淀粉样蛋白25~35 对PC12细胞骨架的毒性作用

曹晶晶范文娟石贞玉鄢明超刘红亮王来邓锦波*

作者信息 +

Effect of amyloid beta-peptide 25-35 neurotoxicity on cytoskeletons of PC12 cells

CAO Jing-jing FAN Wen-juan SHI Zhen-yu YAN Ming-chao LIU Hong-liang WANG Lai DENG Jin-bo*

Author information +

文章历史 +

摘要

目的建立阿尔茨海默病（AD）的细胞模型，并探讨β淀粉样蛋白25~35（Aβ _25~35）对细胞骨架的神经毒性机制。方法采用MTT法检测不同浓度的Aβ _25~35 对PC12细胞存活率的影响，应用4’6-二脒基-2-苯基吲哚（DAPI）染色法和TUNEL法检测细胞凋亡，免疫细胞化学法和鬼笔环肽（phalloidin）染色观察细胞骨架的形态变化。结果 Aβ _25~35经过“老化”处理后，可以明显引发PC12细胞死亡，导致PC12细胞发生凋亡，并具有剂量依赖性（P<0.05）。Aβ _25~35也可以引起细胞骨架解体，同样具有剂量依赖性（P<0.05）。结论 PC12细胞的细胞骨架对Aβ25~35的神经毒性非常敏感，细胞骨架的解体很可能是AD重要的病理学改变，同时Aβ是AD发病机制的关键分子。

Abstract

Objective To establish the cell model of Alzheimer’s disease (AD) and investigate the amyloid beta-peptides _25-35(Aβ _25-35) neurotoxicity to cytoskeleton.Methods PC12 cells were cultured and treated with Aβ _25-35, and cell survival was analyzed by MTT assay. Cell apoptosis was visualized with DAPI staining and TUNEL method. Immunocytochemistry and phalloidin staining were used to label cytoskeletons in PC12 cells. Results Aβ _25-35 obviously induced the PC12 cells death and lead to PC12 cells apoptosis with dose dependency (P<0.05). Aβ _25-35 gave rise to the disintegration of cytoskeletons with dose dependency (P<0.05). Conclusion PC12 cell cytoskeletons are sensitive to Aβ _25-35 neurotoxicity. The disintegration of cytoskeleton probably is the important pathological alteration in AD, and Aβ is a key molecule for AD pathogenesis.

导出引用

曹晶晶范文娟石贞玉鄢明超刘红亮王来邓锦波. β淀粉样蛋白25~35 对PC12细胞骨架的毒性作用[J]. 解剖学报. 2016, 47(4): 469-475 https://doi.org/10.16098/j.issn.0529-1356.2016.04.06

CAO Jing-jing FAN Wen-juan SHI Zhen-yu YAN Ming-chao LIU Hong-liang WANG Lai DENG Jin-bo. Effect of amyloid beta-peptide 25-35 neurotoxicity on cytoskeletons of PC12 cells[J]. Acta Anatomica Sinica. 2016, 47(4): 469-475 https://doi.org/10.16098/j.issn.0529-1356.2016.04.06

参考文献

［1］Dong M，Peng B，Lin X，et al.The prevalence of dementia in the People’s Republic of China：a systematic analysis of 19802004 studies ［J］.Age Ageing，2007，36(6)：619-624.

［2］Alzheimer’s Association.2015 Alzheimer's disease facts and figures ［J］.Alzheimers Dement，2015，11(3)：332-384.

［3］Bayer TA，Wirths O.Focusing the amyloid cascade hypothesis on N-truncated Abeta peptides as drug targets against Alzheimer’s disease ［J］.Acta Neuropathol，2014，127(6)：787-801.

［4］Sadigh-Eteghad S，Sabermarouf B，Majdi A，et al.Amyloid-beta：a crucial factor in Alzheimer's disease ［J］.Med Princ Pract，2015，24(1)：1-10.

［5］Oropesa?vila M，Fernández Vega A，Garrido Maraver J，et al.Emerging roles of apoptotic microtubules during the execution phase of apoptosis ［J］.Cytoskeleton，2015，72(9)：435-446.

［6］Coles CH，Bradke F.Coordinating neuronal actin-microtubule dynamics ［J］.Curr Biol，2015，25(15)：R677-R691.

［7］Rudrabhatla P.Regulation of neuronal cytoskeletal protein phosphorylation in neurodegenerative diseases ［J］.J Alzheimers Dis，2014，41(3)：671-684.

［8］Radford H，Moreno J A，Verity N，et al.PERK inhibition prevents tau-mediated neurodegeneration in a mouse model of frontotemporal dementia ［J］.Acta Neuropathol, 2015，130(5)：633-642.

［9］Wang Y，Wang YJ，Song ShJ，et al.The study of the effect and mechanism of total saponins from Rhizoma Anemarrhenae on Aβ-induced PC12 cell apoptosis ［J］.Chinese Journal of Gerontology，2012，32(13)：2778-2781.(in Chinese)

王艳，王艳杰，宋少江，等.知母总皂苷对β-淀粉样蛋白诱导 PC12 细胞凋亡的抑制作用［J］.中国老年学杂志，2012，32(13)：2778-2781.

［10］Hatok J，Babusikova E，Matakova T，et al.In vitro assays for the evaluation of drug resistance in tumor cells ［J］.Clin Exp Med，2009，9(1)：1-7.

［11］Zhang J，Wu J，Zeng W，et al.Exendin-4, a glucagon-like peptide-1 receptor agonist, inhibits Aβ25-35-induced apoptosis in PC12 cells by suppressing the expression of endoplasmic reticulum stress-related proteins ［J］.Int J Clin Exp Pathol, 2015，8(10)：12784-12792.

［12］Dekkers MPJ，Nikoletopoulou V，Barde YA.Death of developing neurons：new insights and implications for connectivity ［J］.J Cell Biol，2013，203(3)：385-393.

［13］Chen AI，Xiong LJ，Tong Y，et al.The neuroprotective roles of BDNF in hypoxic ischemic brain injury ［J］.Biomed Rep，2013，1(2)：167-176.

［14］Chasseigneaux S，Allinquant B.Functions of Aβ，sAPPα and sAPPβ：similarities and differences ［J］.J Neurochem，2012，120(Suppl 1)：99-108.

［15］Calabrese E J.Amyloid β-peptide：Biphasic dose responses ［J］.Crit Rev Toxicol，2001，31(4-5)：605-606.

［16］Jana A，Pahan K. Fibrillar amyloid-beta peptides kill human primary neurons via NADPH oxidase-mediated activation of neutral sphingomyelinase. Implications for Alzheimer's disease ［J］.J Biol Chem，2004，279(49)：51451-51459.

［17］Martin TFJ，Grishanin RN.PC12 cells as a model for studies of regulated secretion in neuronal and endocrine cells ［J］.Methods Cell Biol，2003，71：267-286.

［18］Loo D T，Copani A，Pike C J，et al.Apoptosis is induced by betaamyloidin culturedcentral nervous system neurons ［J］.Proc Natl Acad Sci USA，1993，90(17)：7951-7955.［19］Roth KA.Caspases，apoptosis，and Alzheimer disease：causation，correlation and confusion ［J］.J Neuropathol Exp Neurol，2001，60(9)：829-838.

［20］Green DR，Llambi F.Cell Death Signaling ［J］.Cold Spring Harb Perspect Biol，2015，7(12)：a006080.

［21］Hardy J.Has the amyloid cascade hypothesis for Alzheimer's disease been proved? ［J］.Curr Alzheimer Res，2006，3(1)：71-73.

［22］Frost B，Bardai FH，Feany MB.Lamin dysfunction mediates neurodegeneration in tauopathies ［J］.Curr Biol，2016，26(1)：129-136.

［23］Lloret A，Fuchsberger T，Giraldo E，et al.Molecular mechanisms linking amyloid β toxicity and Tau hyperphosphorylation in Alzheimer’s disease ［J］.Free Radic Biol Med，2015，83：186-191.

［24］Jin M，Shepardson N，Yang T，et al.Soluble amyloid β-protein dimers isolated from Alzheimer cortex directly induce Tau hyperphosphorylation and neuritic degeneration ［J］.Proc Natl Acad Sci USA，2011，108(14)：5819-5824.

［25］Henriques AG，Oliveira JM，Carvalho LP，et al.Aβ Influences Cytoskeletal Signaling Cascades with Consequences to Alzheimer’s Disease ［J］.Mol Neurobiol，2015，52(3)：1391-1407.

［26］Shen ZY，Xu LY，Li EM，et al.Ezrin, actin and cytoskeleton in apoptosis of esophageal epithelial cells induced by arsenic trioxide ［J］.Int J Mol Med，2003，12(3)：341-347.

［27］Thomas SG，Huang S，Li S，et al.Actin depolymerization is sufficient to induce programmed cell death in self-incompatible pollen ［J］.J Cell Biol，2006，174(2)：221-229.

［28］Ostrovskaya RU，Vakhitova YV，Kuzmina US，et al.Neuroprotective effect of novel cognitive enhancer noopept on AD-related cellular model involves the attenuation of apoptosis and tau hyperphosphorylation ［J］.J Biomed Sci，2014，21(1)：74.