纳米粒子在细胞自噬调控与成像研究中的应用

自噬是真核细胞的基本代谢过程之一,其作用主要是降解和清除细胞内冗余或受损的蛋白质和细胞器,循环利用胞内物质并提供能量.自噬是细胞的一种自我保护机制,同时也与细胞凋亡有着复杂的关联.在病理条件下,自噬水平的非正常上调或抑制与神经退行性疾病及肿瘤、微生物感染等疾病的发生、发展密切相关.因此,阐明自噬的过程和调控机理,并据此攻克人类疾病一直是生物医药领域的研究热点.近年来,随着纳米技术的兴起,纳米材料与自噬的关系也逐渐被揭示,本文总结了国内外有关细胞自噬的最新研究进展,并着重介绍了纳米粒子在调控自噬以及细胞自噬成像方面应用的研究进展,并对其未来的发展方向进行了展望.     

过渡金属离子与神经退行性疾病

介绍过渡金属离子在神经退行性疾病中的作用和生物学意义,以及一种可能的金属作用下的神经退行性疾病致病机理。重点介绍铜离子在老年痴呆症、家族性肌萎侧索硬化症、克雅氏症等疾病的致病机理方面的作用。     

铜锌超氧化物歧化酶的突变与神经退行性紊乱的生物无机化学

本文简单介绍了铜锌超氧化物歧化酶的结构、功能及其相互关系,较为详细地讨论了与神经退行性紊乱,尤其是与肌萎性脊髓侧索硬化症有关的铜锌超氧化物歧化酶突变体的生物无机化学的研究进展,提出由该酶突变而引起的蛋白质局部结构变化和金属离子缺失,以及由此导致的氧化损伤和聚集作用可能是导致神经退行性紊乱的主要原因之一.     

Mn-SOD模拟物及其在神经退行性疾病中的药用前景

本文通过分析神经退行性疾病与线粒体机能障碍、自由基损伤的关系,主要讨论了Mn-SOD模拟物作为自由基清除剂对活性氧化合物的清除机理、药用优势,并总结了近年来有关Mn-SOD模拟物在神经退行性疾病防治方面的研究近况及潜在应用前景。     

基于金属的神经退行性疾病治疗策略*

本文综述了金属离子在神经退行性疾病中的重要作用以及针对该类疾病金属治疗药物的研究进展。以老年痴呆症和帕金森氏症为代表,结合本课题组的初步研究结果,讨论了金属离子在蛋白质聚集与氧化应激反应中的重要作用,暗示金属螯合策略应成为治疗该类疾病的首选策略,并介绍了数种已用于或即将用于临床实验的金属螯合制剂;还介绍了烷基化神经退行性疾病相关蛋白的金属结合位点,可以显著抑制该蛋白质聚集体的形成和活性氧的产生,这可能是继螯合策略后一种更有发展潜力的神经退行性疾病治疗方法。     

O-GlcNAcase抑制剂

O-连接的N-乙酰葡糖胺糖基化修饰 (O-GlcNAcylation) 是一种存在于蛋白质Ser/Thr上的翻译后修饰。与磷酸化相似,它参与细胞内的信号传递,并与神经退行性疾病、Ⅱ型糖尿病、癌症等许多疾病的发病机理密切相关。O-连接的N-乙酰葡糖胺水解酶 (O-GlcNAcase, OGA) 是生物体内唯一水解蛋白质O-GlcNAc修饰的糖苷酶。因此,研究高效、专一的OGA小分子抑制剂是调节细胞中蛋白质O-GlcNAc水平的有效策略,利于阿尔茨海默病等相关神经退行性疾病新型药物的开发。结合本实验室对OGA抑制剂的研究,本文介绍了OGA的结构、催化机理及目前OGA抑制剂的研究进展,讨论了各种抑制剂的构效关系,并对OGA抑制剂的研究前景进行了展望。     

肠道菌群及其代谢产物与神经退行性疾病关系研究进展

神经退行性疾病是一种发生于中枢神经系统,具有高度致残、致死性的疾病,主要发病人群为中老年群体,目前该类疾病的发病机制尚不清楚,没有有效的治疗策略。随着我国老龄化程度的加深,神经退行性疾病对居民身体健康造成严重威胁。肠道菌群作为寄生在胃肠道中的微生物,与人体呈互利共生的关系,对生命健康起到至关重要的作用,神经退行性疾病的发展伴随着肠道菌群及其相关代谢产物的改变。文章综述了肠道菌群及其代谢产物与神经退行性疾病相互影响的机制,并探讨通过肠道菌群治疗神经退行性疾病的潜在价值,以期为神经退行性疾病的治疗提供新的研究方向。     

纳米生物界面对淀粉样蛋白聚集的调控机制研究

本文以阿尔兹海默症的致病多肽β淀粉样蛋白(Aβ)为例,介绍了疾病相关淀粉样蛋白的分子精细结构、组装和聚集过程、聚集体形貌及神经细胞毒性的研究进展,并在此基础上以纳米生物界面对于淀粉样蛋白构象、组装结构、聚集动力学、神经细胞毒性等生物功能的调控机制进行研究.从分子水平上分析和探讨了纳米生物界面与淀粉样蛋白分子或者聚集体的相互作用方式和机理,有助于加深对淀粉样多肽和调节剂之间复杂多样的相互作用方式的理解,对深入了解淀粉样多肽的组装机理和调控机制以及探索治疗神经退行性疾病的药物设计等方面具有较大意义.     

荧光/化学发光探针成像检测超氧阴离子自由基的研究进展

超氧阴离子自由基(O·-2)是细胞内氧气单电子还原后最先产生的一类含氧的高活性物种(活性氧,ROS),与生命过程息息相关.正常稳态浓度的O·-2起重要的信号调控作用,包括细胞的增殖、分化、自噬等.但O·-2浓度的异常,又与癌症、神经退行性疾病、糖尿病等多种疾病的发生发展密切相关.因此,监测O·-2浓度的变化对揭示相关疾病的机理具有至关重要作用.由于荧光成像检测方法具有诸多优势,发展高灵敏、高选择性检测O·-2的荧光探针成为揭示相关疾病发生发展分子机制的关键切入点.近年来,随着荧光显微技术的发展,研究者开发了多种荧光/化学发光探针,实现了对细胞及活体内O·-2水平的可视化监测.本文综述了近五年用于检测O·-2的分子探针、纳米探针、蛋白探针以及化学发光探针的研究进展,并对其发展前景进行了展望.     

它莫西芬诱导乳腺癌MCF-7细胞自噬的比较蛋白质组学研究

自噬(autophagy)广泛存在于真核细胞,贯穿于正常细胞生长发育和生理病理过程.自噬功能的变化与疾病密切相关.     

Why is autophagy important in human diseases?

The process of macroautophagy (referred to hereafter as autophagy), is generally characterized by the prominent formation of autophagic vesicles in the cytoplasm. In the past decades, studies of autophagy have been vastly expanded. As an essential process to maintain cellular homeostasis and functions, autophagy is responsible for the lysosome-mediated degradation of damaged proteins and organelles, and thus misregulation of autophagy can result in a variety of pathological conditions in human beings. Although our understanding of regulatory pathways that control autophagy is still limited, an increasing number of studies have shed light on the importance of autophagy in a wide range of physiological processes and human diseases. The goal of the reviews in the current issue is to provide a general overview of current knowledge on autophagy. The machinery and regulation of autophagy were outlined with special attention to its role in diabetes, neurodegenerative disorders, infectious diseases and cancer.     

Neuronal autophagy and neurodegenerative diseases

Autophagy is a dynamic cellular pathway involved in the turnover of proteins, protein complexes, and organelles through lysosomal degradation. The integrity of postmitotic neurons is heavily dependent on high basal autophagy compared to non-neuronal cells as misfolded proteins and damaged organelles cannot be diluted through cell division. Moreover, neurons contain the specialized structures for intercellular communication, such as axons, dendrites and synapses, which require the reciprocal transport of proteins, organelles and autophagosomes over significant distances from the soma. Defects in autophagy affect the intercellular communication and subsequently, contributing to neurodegeneration. The presence of abnormal autophagic activity is frequently observed in selective neuronal populations afflicted in common neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis. These observations have provoked controversy regarding whether the increase in autophagosomes observed in the degenerating neurons play a protective role or instead contribute to pathogenic neuronal cell death. It is still unknown what factors may determine whether active autophagy is beneficial or pathogenic during neurodegeneration. In this review, we consider both the normal and pathophysiological roles of neuronal autophagy and its potential therapeutic implications for common neurodegenerative diseases.     

Quantum dots protect against MPP+-induced neurotoxicity in a cell model of Parkinson’s disease through autophagy induction

Autophagy is a basic cellular process that decomposes damaged organelles and aberrant proteins. Dysregulation of autophagy is implicated in pathogenesis of neurodegenerative disorders, including Parkinson's disease(PD). Pharmacological compounds that stimulate autophagy can provide neuroprotection in models of PD. Nanoparticles have emerged as regulators of autophagy and have been tested in adjuvant therapy for diseases. In this present study, we explore the effects of quantum dots(QDs) that can induce autophagy in a cellular model of Parkinson's disease. Cd Te/Cd S/Zn S QDs protect differentiated rat pheochromocytoma PC12 cells from MPP+-induced cell damage, including reduced viability, apoptosis and accumulation of α-Synuclein, a characteristic protein of PD. The protective function of QDs is autophagy-dependent. In addition, we investigate the interaction between quantum dots and autophagic pathways and identify beclin1 as an essential factor for QDs-induced autophagy. Our results reveal new promise of QDs in the theranostic of neurodegenerative diseases.     

Autophagy regulation by acetylation—implications for neurodegenerative diseases

Posttranslational modifications of proteins, such as acetylation, are essential for the regulation of diverse physiological processes, including metabolism, development and aging. Autophagy is an evolutionarily conserved catabolic process that involves the highly regulated sequestration of intracytoplasmic contents in double-membrane vesicles called autophagosomes, which are subsequently degraded after fusing with lysosomes. The roles and mechanisms of acetylation in autophagy control have emerged only in the last few years. In this review, we describe key molecular mechanisms by which previously identified acetyltransferases and deacetylases regulate autophagy. We highlight how p300 acetyltransferase controls mTORC1 activity to regulate autophagy under starvation and refeeding conditions in many cell types. Finally, we discuss how altered acetylation may impact various neurodegenerative diseases in which many of the causative proteins are autophagy substrates. These studies highlight some of the complexities that may need to be considered by anyone aiming to perturb acetylation under these conditions.Subject terms: Acetylation, Alzheimer''s disease     

Autophagy-regulating small molecules and their therapeutic applications

Autophagy or self-eating is a complicated cellular process that is involved in protein and organelle digestion occurring via a lysosome-dependent pathway. This process is of great importance in maintaining normal cellular homeostasis. However, disruption of autophagy is closely associated with various human diseases such as cancer, neurodegenerative disorders, heart disease and pathogen infection. Therefore, small molecules that modulate autophagy can be employed to dissect this complex process and ultimately could have high potential for the treatment of a variety of diseases. This critical review discusses general aspects of autophagy, autophagy-associated diseases and autophagy regulators for biological research and therapeutic applications (207 references).     

Dioscin-Mediated Autophagy Alleviates MPP+-Induced Neuronal Degeneration: An In Vitro Parkinson’s Disease Model

Autophagy is a cellular homeostatic process by which cells degrade and recycle their malfunctioned contents, and impairment in this process could lead to Parkinson’s disease (PD) pathogenesis. Dioscin, a steroidal saponin, has induced autophagy in several cell lines and animal models. The role of dioscin-mediated autophagy in PD remains to be investigated. Therefore, this study aims to investigate the hypothesis that dioscin-regulated autophagy and autophagy-related (ATG) proteins could protect neuronal cells in PD via reducing apoptosis and enhancing neurogenesis. In this study, the 1-methyl-4-phenylpyridinium ion (MPP⁺) was used to induce neurotoxicity and impair autophagic flux in a human neuroblastoma cell line (SH-SY5Y). The result showed that dioscin pre-treatment counters MPP⁺-mediated autophagic flux impairment and alleviates MPP⁺-induced apoptosis by downregulating activated caspase-3 and BCL2 associated X, apoptosis regulator (Bax) expression while increasing B-cell lymphoma 2 (Bcl-2) expression. In addition, dioscin pre-treatment was found to increase neurotrophic factors and tyrosine hydroxylase expression, suggesting that dioscin could ameliorate MPP⁺-induced degeneration in dopaminergic neurons and benefit the PD model. To conclude, we showed dioscin’s neuroprotective activity in neuronal SH-SY5Y cells might be partly related to its autophagy induction and suppression of the mitochondrial apoptosis pathway.     

Exercise-induced FNDC5/irisin protects nucleus pulposus cells against senescence and apoptosis by activating autophagy

Intervertebral disc degeneration (IVDD) is a major cause of low back pain (LBP), and excessive senescence and apoptosis of nucleus pulposus (NP) cells are major pathological changes in IVDD. Physical exercise could effectively delay the process of intervertebral disc degeneration; however, its mechanism is still largely unknown. Irisin is an exercise-induced myokine released upon cleavage of the membrane-bound precursor protein fibronectin type III domain-containing protein 5 (FNDC5), and its levels increase after physical exercise. Here, we show that after physical exercise, FNDC5/irisin levels increase in the circulation and NP, senescence and apoptosis are reduced, autophagy is activated in NP tissue, and the progression of IVDD is delayed. Conversely, after knocking out FNDC5, the benefits of physical exercise are compromised. Moreover, the overexpression of FNDC5 in NP tissue effectively alleviated the degeneration of the intervertebral disc (IVD) in rats. By showing that FNDC5/irisin is an important mediator of the beneficial effects of physical exercise in the IVDD model, the study proposes FNDC5/irisin as a novel agent capable of activating autophagy and protecting NP from senescence and apoptosis.Subject terms: Endocrinology, Diseases, Bone     

The roles of the inhibitory autophagy regulator Rubicon in the heart: A new therapeutic target to prevent cardiac cell death

Autophagy contributes to the maintenance of cardiac homeostasis. The level of autophagy is dynamically altered in heart disease. Although autophagy is a promising therapeutic target, only a few selective autophagy activator candidates have been reported thus far. Rubicon is one of the few endogenous negative regulators of autophagy and a potential target for autophagy-inducing therapeutics. Rubicon was initially identified as a component of the Class III PI3K complex, and it has multiple functions, not only in canonical autophagy but also in endosomal trafficking and inflammatory responses. This review summarizes the molecular action of Rubicon in canonical and noncanonical autophagy. We discuss the roles of Rubicon in cardiac stress and the therapeutic potential of Rubicon in cardiac diseases through its modulation of autophagy.Subject terms: Macroautophagy, Mechanisms of disease     

Degradation of misfolded proteins in neurodegenerative diseases: therapeutic targets and strategies

Mammalian cells remove misfolded proteins using various proteolytic systems, including the ubiquitin (Ub)-proteasome system (UPS), chaperone mediated autophagy (CMA) and macroautophagy. The majority of misfolded proteins are degraded by the UPS, in which Ub-conjugated substrates are deubiquitinated, unfolded and cleaved into small peptides when passing through the narrow chamber of the proteasome. The substrates that expose a specific degradation signal, the KFERQ sequence motif, can be delivered to and degraded in lysosomes via the CMA. Aggregation-prone substrates resistant to both the UPS and the CMA can be degraded by macroautophagy, in which cargoes are segregated into autophagosomes before degradation by lysosomal hydrolases. Although most misfolded and aggregated proteins in the human proteome can be degraded by cellular protein quality control, some native and mutant proteins prone to aggregation into β-sheet-enriched oligomers are resistant to all known proteolytic pathways and can thus grow into inclusion bodies or extracellular plaques. The accumulation of protease-resistant misfolded and aggregated proteins is a common mechanism underlying protein misfolding disorders, including neurodegenerative diseases such as Huntington''s disease (HD), Alzheimer''s disease (AD), Parkinson''s disease (PD), prion diseases and Amyotrophic Lateral Sclerosis (ALS). In this review, we provide an overview of the proteolytic pathways in neurons, with an emphasis on the UPS, CMA and macroautophagy, and discuss the role of protein quality control in the degradation of pathogenic proteins in neurodegenerative diseases. Additionally, we examine existing putative therapeutic strategies to efficiently remove cytotoxic proteins from degenerating neurons.