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Acyl-CoA Synthetase Long-Chain Isoenzymes in Kidney Diseases: Mechanistic Insights and Therapeutic Implications 肾脏疾病中的酰基-CoA 合成酶长链异酶:机理认识和治疗意义
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-30 DOI: 10.1002/cbf.4114
Swati Mishra, Vishwadeep Shelke, Anil Bhanudas Gaikwad

Long-chain acyl-CoA synthetases (ACSLs) are pivotal enzymes in fatty acid metabolism, essential for maintaining cellular homeostasis and energy production. Recent research has uncovered their significant involvement in the pathophysiology of various kidney diseases, including acute kidney injury (AKI), chronic kidney disease (CKD), diabetic kidney disease (DKD), and renal cell carcinoma (RCC). While ACSL1, ACSL3, ACSL4, and ACSL5 have been extensively studied for their roles in processes such as ferroptosis, lipid peroxidation, renal fibrosis, epithelial-mesenchymal transition, and tumor progression, the role of ACSL6 in kidney diseases remain largely unexplored. Notably, these isoenzymes exhibit distinct functions in different kidney diseases. Therefore, to provide a comprehensive understanding of their involvement, this review highlights the molecular pathways influenced by ACSLs and their roles in modulating cell death, inflammation, and fibrosis during kidney disease progression. By examining these mechanisms in detail, this review underscores the potential of ACSLs as biomarkers and therapeutic targets, advocating for further research to elucidate the precise roles of individual ACSL isoenzymes in kidney disease progression. Understanding these mechanisms opens new avenues for developing targeted interventions and improving therapeutic outcomes for patients with kidney diseases.

长链酰基-CoA 合成酶(ACSL)是脂肪酸代谢中的关键酶,对维持细胞平衡和能量生产至关重要。最近的研究发现,它们在急性肾损伤(AKI)、慢性肾病(CKD)、糖尿病肾病(DKD)和肾细胞癌(RCC)等多种肾脏疾病的病理生理学中发挥着重要作用。ACSL1、ACSL3、ACSL4 和 ACSL5 在铁变态反应、脂质过氧化、肾脏纤维化、上皮-间质转化和肿瘤进展等过程中的作用已被广泛研究,但 ACSL6 在肾脏疾病中的作用在很大程度上仍未被探索。值得注意的是,这些同工酶在不同的肾脏疾病中表现出不同的功能。因此,为了全面了解它们的参与情况,本综述重点介绍了受 ACSLs 影响的分子通路及其在肾脏疾病进展过程中调节细胞死亡、炎症和纤维化的作用。通过详细研究这些机制,本综述强调了 ACSLs 作为生物标志物和治疗靶点的潜力,提倡进一步研究以阐明单个 ACSL 同工酶在肾病进展中的确切作用。了解这些机制为开发有针对性的干预措施和改善肾病患者的治疗效果开辟了新途径。
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引用次数: 0
Levilactobacillus brevis IBARAKI-TS3 Isolated From Pickles Promotes Production of Interleukin-10 via Toll-Like Receptor 2 in Human M2 Macrophages 从泡菜中分离出的左旋乳杆菌 IBARAKI-TS3 通过 Toll-Like 受体 2 促进人类 M2 巨噬细胞产生白细胞介素-10
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-29 DOI: 10.1002/cbf.4110
Keisuke Tobita, Satoru Iwasa

M2 macrophages play an important role in food allergy. Several studies have reported that lactic acid bacteria isolated from pickles exert antiallergic effects. We investigated the effects of several strains of lactic acid bacteria on the immune function of M2 macrophages. M2 macrophages differentiated from THP-1 cell line by interleukin-4 (IL-4) and IL-13 strongly expressed CD163, CD206, and HMOX1 mRNA. Levilactobacillus brevis IBARAKI-TS3 (IBARAKI-TS3) isolated from pickles was identified as a lactic acid bacterium that enhances the expressions of IL-10 and EBI3 mRNA in M2 macrophages. IBARAKI-TS3 induced the expression of genes involved in Toll-like receptor (TLR) signaling, such as IRAK, mitogen-activated protein kinases (MAPKs), and NF-κB mRNA. IBARAKI-TS3–induced IL-10 production was suppressed by anti-TLR2–neutralizing antibodies. Furthermore, the IBARAKI-TS3–induced increase in IL-10 levels was significantly reduced in TLR2–knockdown M2 macrophages compared to M2 macrophages. These results suggest that IBARAKI-TS3 promotes of IL-10 production via TLR2 in M2 macrophages.

M2 巨噬细胞在食物过敏中发挥着重要作用。一些研究报告称,从腌菜中分离出的乳酸菌具有抗过敏作用。我们研究了几种乳酸菌对 M2 巨噬细胞免疫功能的影响。通过白细胞介素-4(IL-4)和 IL-13 从 THP-1 细胞系分化出的 M2 巨噬细胞强烈表达 CD163、CD206 和 HMOX1 mRNA。从腌菜中分离出的Levilactobacillus brevis IBARAKI-TS3(IBARAKI-TS3)被鉴定为一种乳酸菌,能增强M2巨噬细胞中IL-10和EBI3 mRNA的表达。IBARAKI-TS3可诱导参与Toll样受体(TLR)信号转导的基因表达,如IRAK、丝裂原活化蛋白激酶(MAPKs)和NF-κB mRNA。抗TLR2-中和抗体抑制了IBARAKI-TS3诱导的IL-10产生。此外,TLR2-敲除的 M2 巨噬细胞与 M2 巨噬细胞相比,IBARAKI-TS3 诱导的 IL-10 水平升高明显降低。这些结果表明,IBARAKI-TS3 可通过 TLR2 促进 M2 巨噬细胞产生 IL-10。
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引用次数: 0
The humoral immune landscape in Parkinson's disease: Unraveling antibody and B cell changes 帕金森病的体液免疫状况:揭示抗体和 B 细胞的变化。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-27 DOI: 10.1002/cbf.4109
Zahra Baridjavadi, Mahmoud Mahmoudi, Narges Abdollahi, Negar Ebadpour, Samaneh mollazadeh, Dariush Haghmorad, Seyed-Alireza Esmaeili

Parkinson's disease (PD) is a complex neurodegenerative disorder characterized by the accumulation of α-synuclein (α-syn) in the brain and progressive loss of dopaminergic neurons in the substantia nigra (SN) region of the brain. Although the role of neuroinflammation and cellular immunity in PD has been extensively studied, the involvement of humoral immunity mediated by antibodies and B cells has received less attention. This article provides a comprehensive review of the current understanding of humoral immunity in PD. Here, we discuss alterations in B cells in PD, including changes in their number and phenotype. Evidence mostly indicates a decrease in the quantity of B cells in PD, accompanied by a shift in the population from naïve to memory cells. Furthermore, the existence of autoantibodies that target several antigens in PD has been investigated (i.e., anti-α-syn autoantibodies, anti-glial-derived antigen antibodies, anti-Tau antibodies, antineuromelanin antibodies, and antibodies against the renin-angiotensin system). Several autoantibodies are generated in PD, which may either provide protection or have harmful effects on disease progression. Furthermore, we have reviewed studies focusing on the utilization of antibodies as a potential treatment for PD, both in animal and clinical trials. This review sheds light on the intricate interplay between antibodies and the pathological processes in PD, including complement system activation.

帕金森病(PD)是一种复杂的神经退行性疾病,其特征是大脑中α-突触核蛋白(α-syn)的积累和大脑黑质(SN)区域多巴胺能神经元的逐渐丧失。尽管神经炎症和细胞免疫在帕金森病中的作用已被广泛研究,但由抗体和B细胞介导的体液免疫的参与却较少受到关注。本文全面回顾了目前对帕金森病中体液免疫的认识。在此,我们将讨论帕金森病中 B 细胞的变化,包括其数量和表型的变化。大多数证据表明,帕金森病患者的B细胞数量减少,同时出现从幼稚细胞到记忆细胞的转变。此外,研究还发现,在帕金森病中存在针对多种抗原的自身抗体(即抗α-syn自身抗体、抗胶质衍生抗原抗体、抗Tau抗体、抗神经美兰宁抗体和抗肾素-血管紧张素系统抗体)。在帕金森病中会产生多种自身抗体,这些抗体既可能提供保护,也可能对疾病进展产生有害影响。此外,我们还回顾了在动物实验和临床试验中将抗体作为一种潜在治疗方法的研究。这篇综述揭示了抗体与帕金森病病理过程(包括补体系统激活)之间错综复杂的相互作用。
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引用次数: 0
Role of osteokines in atherosclerosis 骨激酶在动脉粥样硬化中的作用
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-18 DOI: 10.1002/cbf.4107
Yi-Fan Liu, Yuan Tian, Xiao-Fang Chen, Chi Zhang, Liang Huang

Despite their diverse physiologies and roles, the heart, skeletal muscles, and smooth muscles all derive from a common embryonic source as bones. Moreover, bone tissue, skeletal and smooth muscles, and the heart share conserved signaling pathways. The maintenance of skeletal health is precisely regulated by osteocytes, osteoblasts, and osteoclasts through coordinated secretion of bone-derived factors known as osteokines. Increasing evidence suggests the involvement of osteokines in regulating atherosclerotic vascular disease. Therefore, this review aims to examine the evidence for the role of osteokines in atherosclerosis development and progression comprehensively. Specifically discussed are extensively studied osteokines in atherosclerosis such as osteocalcin, osteopontin, osteoprotegerin, and fibroblast growth factor 23. Additionally, we highlighted the effects of exercise on modulating these key regulators derived from bone tissue metabolism. We believe that gaining an enhanced understanding of how osteocalcin contributes to the process of atherosclerosis will enable us to develop targeted and comprehensive therapeutic strategies against diseases associated with its progression.

尽管心脏、骨骼肌和平滑肌的生理结构和作用各不相同,但它们都源自骨骼这一共同的胚胎来源。此外,骨组织、骨骼肌、平滑肌和心脏共享保守的信号通路。骨骼健康的维持是由骨细胞、成骨细胞和破骨细胞通过协调分泌骨源性因子(即骨激素)来精确调控的。越来越多的证据表明,骨生成素参与了动脉粥样硬化性血管疾病的调控。因此,本综述旨在全面研究骨促性因子在动脉粥样硬化发生和发展过程中发挥作用的证据。具体讨论了在动脉粥样硬化中被广泛研究的骨调理因子,如骨钙素、骨通蛋白、骨保护蛋白和成纤维细胞生长因子 23。此外,我们还强调了运动对调节这些来自骨组织代谢的关键调节因子的影响。我们相信,进一步了解骨钙素是如何促进动脉粥样硬化过程的,将有助于我们针对与动脉粥样硬化进展相关的疾病制定有针对性的综合治疗策略。
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引用次数: 0
Myostatin's marvels: From muscle regulator to diverse implications in health and disease Myostatin 的奇迹:从肌肉调节器到对健康和疾病的各种影响
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-14 DOI: 10.1002/cbf.4106
Sonakshi Sharma, Amol S. Patil

Myostatin, a member of the transforming growth factor-β superfamily, is a pivotal regulator of skeletal muscle growth in mammals. Its discovery has sparked significant interest due to its multifaceted roles in various physiological processes and its potential therapeutic implications. This review explores the diverse functions of myostatin in skeletal muscle development, maintenance and pathology. We delve into its regulatory mechanisms, including its interaction with other signalling pathways and its modulation by various factors such as microRNAs and mechanical loading. Furthermore, we discuss the therapeutic strategies aimed at targeting myostatin for the treatment of muscle-related disorders, including cachexia, muscular dystrophy and heart failure. Additionally, we examine the impact of myostatin deficiency on craniofacial morphology and bone development, shedding light on its broader implications beyond muscle biology. Through a comprehensive analysis of the literature, this review underscores the importance of further research into myostatin's intricate roles and therapeutic potential in human health and disease.

Myostatin是转化生长因子-β超家族的成员,是哺乳动物骨骼肌生长的关键调节因子。由于它在各种生理过程中的多方面作用及其潜在的治疗意义,它的发现引发了人们的极大兴趣。这篇综述探讨了肌节蛋白在骨骼肌发育、维持和病理过程中的多种功能。我们深入探讨了它的调控机制,包括它与其他信号通路的相互作用,以及它受各种因素(如 microRNA 和机械负荷)的调节。此外,我们还讨论了针对肌节蛋白的治疗策略,以治疗肌肉相关疾病,包括恶病质、肌肉萎缩症和心力衰竭。此外,我们还研究了肌生长抑素缺乏对颅面形态和骨骼发育的影响,从而揭示了肌生长抑素在肌肉生物学之外的更广泛意义。通过对文献的全面分析,本综述强调了进一步研究肌节蛋白在人类健康和疾病中的复杂作用和治疗潜力的重要性。
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引用次数: 0
Microfluidic device: A versatile biosensor platform to multiplex aptamer-based detection of malaria biomarkers 微流控装置:多功能生物传感器平台,可基于多通道适配体检测疟疾生物标记物。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-08 DOI: 10.1002/cbf.4104
Adewoyin M. Ogunmolasuyi, Mary A. Adewoyin

Plasmodium falciparum malaria remains a dominant infectious disease that affects Africa than the rest of the world, considering its associated cases and death rates. It's a febrile illness that produces several reliable biomarkers, for example, P. falciparum lactate dehydrogenase (PfLDH), P. falciparum Plasmodium glutamate dehydrogenase (PfGDH), and P. falciparum histidine-rich proteins (HRP-II) in blood circulatory system that can easily be employed as targets in rapid diagnostic tests (RDTs). In recent times, several DNA aptamers have been developed via SELEX technology to detect some specific malaria biomarkers (PfLDH, PvLDH, HRP-II, PfGDH) in a biosensor mode with good binding affinity properties to overcome the trend of cross-reactivity, limited sensitivity and stability problems that have been observed with immunodiagnostics. In this review, we summarized existing diagnostic methods and relevant biomarkers to suggest promising approaches to develop sensitive and species-specific multiplexed diagnostic devices enabling effective detection of malaria in complex biological matrices and surveillance in the endemic region.

恶性疟原虫疟疾仍然是影响非洲的主要传染病,其发病率和死亡率高于世界其他地区。恶性疟原虫疟疾是一种发热性疾病,会在血液循环系统中产生多种可靠的生物标志物,例如恶性疟原虫乳酸脱氢酶(PfLDH)、恶性疟原虫谷氨酸脱氢酶(PfGDH)和恶性疟原虫富组氨酸蛋白(HRP-II),这些标志物很容易被用作快速诊断检测(RDT)的靶标。近来,人们通过 SELEX 技术开发了几种 DNA 嵌合体,以生物传感器模式检测一些特定的疟疾生物标志物(PfLDH、PvLDH、HRP-II、PfGDH),这些嵌合体具有良好的结合亲和性,克服了免疫诊断技术中存在的交叉反应趋势、灵敏度有限和稳定性问题。在这篇综述中,我们总结了现有的诊断方法和相关的生物标记物,提出了开发灵敏的、物种特异的多重诊断设备的可行方法,以便在复杂的生物基质中有效检测疟疾,并在疟疾流行地区进行监测。
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引用次数: 0
Anion exchanger1 (AE1/SLC4A1) function is impaired in red blood cells from prediabetic subjects: Potential benefits of finger lime (Citrus australasica, Faustrime cultivar) juice extract 糖尿病前期患者的红细胞中阴离子交换器1(AE1/SLC4A1)功能受损:指柠檬(Citrus australasica, Faustrime cultivar)果汁提取物的潜在益处。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-02 DOI: 10.1002/cbf.4105
Alessia Remigante, Sara Spinelli, Lucrezia Gambardella, Elisabetta Straface, Giovanna Cafeo, Marina Russo, Daniele Caruso, Paola Dugo, Silvia Dossena, Angela Marino, Rossana Morabito

Prediabetes is a risk state that defines a high chance of developing diabetes and cardiovascular disease. Oxidative stress mediated by hyperglycemia-induced production of reactive species could play a crucial role in this context. In the present study, we investigated whether the anion exchange capability mediated by AE1 (SLC4A1), which is sensitive to oxidative stress, was altered in human red blood cells (RBCs) obtained from prediabetic volunteers. In addition, we assessed the precise composition of bioactive compounds and the potential benefits of finger lime juice extract (Citrus australasica, Faustrime cultivar) in counteracting oxidative stress-related functional alterations. Human RBCs from normal and prediabetic volunteers were incubated with 50 µg/mL juice extract for 2 h at 25°C. Juice extract restored alterations of the anion exchange capability mediated by AE1 and prevented the structural rearrangements of AE1 and α/β-spectrin in prediabetic RBCs. AE1 functional and structural alterations were not associated with an increase in lipid peroxidation or protein oxidation at the level of the plasma membrane. An increased production of intracellular ROS, which provoked the oxidation of hemoglobin to methemoglobin, both reverted by juice extract, was instead observed. Importantly, juice extract also induced a reduction in glycated hemoglobin levels in prediabetic RBCs. Finally, juice extract blunted the overactivation of the endogenous antioxidant enzymes catalase and superoxide dismutase and prevented glutathione depletion in prediabetic RBCs. These findings contribute to clarifying cellular and molecular mechanisms related to oxidative stress and glycation events that may influence RBC and systemic homeostasis in prediabetes, identify AE1 as a sensitive biomarker of RBC structural and function alterations in prediabetes and propose finger lime juice extract as a natural antioxidant for the treatment and/or prevention of the complications associated with the prediabetic condition.

糖尿病前期是一种风险状态,决定了患糖尿病和心血管疾病的几率很高。在这种情况下,由高血糖诱导产生的活性物质所介导的氧化应激可能起着至关重要的作用。在本研究中,我们调查了从糖尿病前期志愿者体内获得的人类红细胞(RBCs)中由 AE1(SLC4A1)介导的阴离子交换能力是否发生了改变,AE1 对氧化应激很敏感。此外,我们还评估了指柠檬汁提取物(Citrus australasica,Faustrime 栽培品种)中生物活性化合物的精确成分及其在抵消氧化应激相关功能改变方面的潜在益处。将正常和糖尿病前期志愿者的人体红细胞与 50 µg/mL 的果汁提取物在 25°C 下培养 2 小时。果汁提取物恢复了由 AE1 介导的阴离子交换能力的改变,并防止了糖尿病前期红细胞中 AE1 和 α/β-spectrin 的结构重排。AE1 功能和结构的改变与质膜水平的脂质过氧化或蛋白质氧化的增加无关。相反,观察到细胞内 ROS 生成增加,导致血红蛋白氧化成高铁血红蛋白,而果汁提取物可使这两种现象恢复。重要的是,果汁提取物还能降低糖尿病前期红细胞的糖化血红蛋白水平。最后,果汁提取物能减弱内源性抗氧化酶过氧化氢酶和超氧化物歧化酶的过度活化,并防止糖尿病前期红细胞中谷胱甘肽的耗竭。这些发现有助于阐明与氧化应激和糖化事件有关的细胞和分子机制,这些机制可能会影响糖尿病前期红细胞和全身的平衡,确定 AE1 是糖尿病前期红细胞结构和功能改变的敏感生物标志物,并建议将指柠檬汁提取物作为一种天然抗氧化剂,用于治疗和/或预防与糖尿病前期相关的并发症。
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引用次数: 0
The interaction between cannabinoids and long-term synaptic plasticity: A survey on memory formation and underlying mechanisms 大麻素与长期突触可塑性之间的相互作用:记忆形成和内在机制调查。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-01 DOI: 10.1002/cbf.4100
Maryam Azarfarin, Tahereh Ghadiri, Masoomeh Dadkhah, Sajad Sahab-Negah

Synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD), is an essential phenomenon in memory formation as well as maintenance along with many other cognitive functions, such as those needed for coping with external stimuli. Synaptic plasticity consists of gradual changes in the biochemistry and morphology of pre- and postsynaptic neurons, particularly in the hippocampus. Consuming marijuana as a primary source of exocannabinoids immediately impairs attention and working memory-related tasks. Evidence regarding the effects of cannabinoids on LTP and memory is contradictory. While cannabinoids can affect a variety of specific cannabinoid receptors (CBRs) and nonspecific receptors throughout the body and brain, they exert miscellaneous systemic and local cerebral effects. Given the increasing use of cannabis, mainly among the young population, plus its potential adverse long-term effects on learning and memory processes, it could be a future global health challenge. Indeed, the impact of cannabinoids on memory is multifactorial and depends on the dosage, timing, formula, and route of consumption, plus the background complex interaction of the endocannabinoids system with other cerebral networks. Herein, we review how exogenously administrated organic cannabinoids, CBRs agonists or antagonists, and endocannabinoids can affect LTP and synaptic plasticity through various receptors in interaction with other cerebral pathways and primary neurotransmitters.

突触可塑性,包括长期电位(LTP)和长期抑制(LTD),是记忆形成和维持的重要现象,也是许多其他认知功能(如应对外部刺激所需的功能)的重要组成部分。突触可塑性包括突触前和突触后神经元的生物化学和形态的逐渐变化,尤其是在海马体中。作为外大麻素的主要来源,吸食大麻会立即损害注意力和与工作记忆相关的任务。有关大麻素对 LTP 和记忆影响的证据相互矛盾。虽然大麻素可以影响全身和大脑的各种特异性大麻素受体(CBRs)和非特异性受体,但它们对全身和局部大脑产生多种影响。鉴于大麻的使用日益增多,主要是在年轻人群中,加上大麻对学习和记忆过程可能产生的长期不利影响,大麻可能成为未来全球健康的一个挑战。事实上,大麻素对记忆的影响是多因素的,取决于服用的剂量、时间、配方和途径,以及内源性大麻素系统与其他大脑网络之间复杂的相互作用背景。在此,我们回顾了外源性给药的有机大麻素、CBRs 激动剂或拮抗剂以及内源性大麻素如何通过各种受体与其他大脑通路和主要神经递质相互作用,影响 LTP 和突触可塑性。
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引用次数: 0
Creation of an in vitro model of GM1 gangliosidosis by CRISPR/Cas9 knocking-out the GLB1 gene in SH-SY5Y human neuronal cell line 通过 CRISPR/Cas9 基因敲除 SH-SY5Y 人类神经细胞系中的 GLB1 基因,创建 GM1 神经节苷脂病的体外模型。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-30 DOI: 10.1002/cbf.4102
Kamran Hosseini, Jafar Fallahi, Hadi Aligholi, Zahra Heidari, Elham Nadimi, Fatemeh Safari, Mohsen Sisakht, Amir Atapour, Sahar Khajeh, Seyed Mohammad Bagher Tabei, Vahid Razban

GM1 gangliosidosis is one type of hereditary error of metabolism that occurs due to the absence or reduction of β-galactosidase enzyme content in the lysosome of cells, including neurons. In vitro, the use of neural cell lines could facilitate the study of this disease. By creating a cell model of GM1 gangliosidosis on the SH-SY5Y human nerve cell line, it is possible to understand the main role of this enzyme in breaking down lipid substrate and other pathophysiologic phenomena this disease. To knock-out the human GLB1 gene, guides targeting exons 14 and 16 of the GLB1 gene were designed using the CRISPOR and CHOP-CHOP websites, and high-efficiency guides were selected for cloning in the PX458 vector. After confirming the cloning, the vectors were transformed into DH5α bacteria and then the target vector was extracted and transfected into human nerve cells (SH-SY5Y cell line) by electroporation. After 48 h, GFP+ cells were sorted using the FACS technique and homozygous (compound heterozygous) single cells were isolated using the serial dilution method and sequencing was done to confirm them. Finally, gap PCR tests, X-gal and Periodic acid-Schiff (PAS) staining, and qPCR were used to confirm the knock-out of the human GLB1 gene. Additionally, RNA sequencing data analysis from existing data of the Gene Expression Omnibus (GEO) was used to find the correlation of GLB1 with other genes, and then the top correlated genes were tested for further evaluation of knock-out effects. The nonviral introduction of two guides targeting exons 14 and 16 of the GLB1 gene into SH-SY5Y cells led to the deletion of a large fragment with a size of 4.62 kb. In contrast to the non-transfected cell, X-gal staining resulted in no blue color in GLB1 gene knock-out cells indicating the absence of β-galactosidase enzyme activity in these cells. Real-time PCR (qPCR) results confirmed the RNA-Seq analysis outcomes on the GEO data set and following the GLB1 gene knock-out, the expression of its downstream genes, NEU1 and CTSA, has been decreased. It has been also shown that the downregulation of GLB1-NEU1-CTSA complex gene was involved in suppressed proliferation and invasion ability of knock-out cells. This study proved that using dual guide RNA can be used as a simple and efficient tool for targeting the GLB1 gene in nerve cells and the knockout SH-SY5Y cells can be used as a model investigation of basic and therapeutic surveys for GM1 gangliosidosis disease.

GM1神经节苷脂病是一种遗传性代谢错误,是由于包括神经元在内的细胞溶酶体中的β-半乳糖苷酶含量缺乏或减少而导致的。在体外,使用神经细胞系可以促进对这种疾病的研究。通过在 SH-SY5Y 人类神经细胞系上建立 GM1 神经节苷脂病的细胞模型,可以了解这种酶在分解脂质底物中的主要作用以及这种疾病的其他病理生理现象。为了敲除人GLB1基因,利用CRISPOR和CHOP-CHOP网站设计了针对GLB1基因第14和16外显子的向导,并选择高效向导克隆到PX458载体中。确认克隆成功后,将载体转化到 DH5α 细菌中,然后提取目的载体并通过电穿孔转染人神经细胞(SH-SY5Y 细胞系)。48 小时后,用 FACS 技术对 GFP+ 细胞进行分选,并用系列稀释法分离出同种异型(复合杂合)单细胞,然后进行测序确认。最后,利用间隙 PCR 检测、X-gal 和周期酸-Schiff(PAS)染色以及 qPCR 来确认人类 GLB1 基因的敲除。此外,还利用基因表达总库(Gene Expression Omnibus,GEO)现有数据中的 RNA 测序数据分析,找出 GLB1 与其他基因的相关性,然后检测相关性最高的基因,进一步评估基因敲除效果。将两个靶向 GLB1 基因第 14 和 16 号外显子的导引片段以非病毒方式导入 SH-SY5Y 细胞后,删除了一个大小为 4.62 kb 的大片段。与未转染细胞相比,GLB1 基因敲除细胞的 X-gal 染色结果显示没有蓝色,这表明这些细胞中没有 β-半乳糖苷酶活性。实时 PCR(qPCR)结果证实了 GEO 数据集上的 RNA-Seq 分析结果,GLB1 基因敲除后,其下游基因 NEU1 和 CTSA 的表达量减少。研究还表明,GLB1-NEU1-CTSA 复合物基因的下调参与抑制了基因敲除细胞的增殖和侵袭能力。这项研究证明,使用双向导 RNA 可以作为一种简单而有效的工具来靶向神经细胞中的 GLB1 基因,而基因敲除的 SH-SY5Y 细胞可作为研究 GM1 神经节苷脂病基础和治疗的模型。
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引用次数: 0
A comprehensive overview of radiation therapy impacts of various cancer treatments and pivotal role in the immune system 全面介绍放射治疗对各种癌症治疗的影响以及在免疫系统中的关键作用。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-29 DOI: 10.1002/cbf.4103
Dhasarathdev Srinivasan, Rajasekaran Subbarayan, Nityanand Srivastava, Arunkumar Radhakrishnan, Pooja Narain Adtani, Ankush Chauhan, Loganathan Krishnamoorthy

The cancer treatment landscape is significantly evolving, focusing on advanced radiation therapy methods to maximize effectiveness and minimize the adverse effects. Recognized as a pivotal component in cancer and disease treatment, radiation therapy (RT) has drawn attention in recent research that delves into its intricate interplay with inflammation and the immune response. This exploration unveils the underlying processes that significantly influence treatment outcomes. In this context, the potential advantages of combining bronchoscopy with RT across diverse clinical scenarios, alongside the targeted impact of brachytherapy, are explored. Concurrently, radiation treatments serve multifaceted roles such as DNA repair, cell elimination, and generating immune stress signaling molecules known as damage-associated molecular patterns, elucidating their effectiveness in treating various diseases. External beam RT introduces versatility by utilizing particles such as photons, electrons, protons, or carbon ions, each offering distinct advantages. Advanced RT techniques contribute to the evolving landscape, with emerging technologies like FLASH, spatially fractionated RT, and others poised to revolutionize the field. The comprehension of RT, striving for improved treatment outcomes, reduced side effects, and facilitating personalized and innovative treatments for cancer and noncancer patients. After navigating these advancements, the goal is fixed to usher in a new era in which RT is a cornerstone of precision and effectiveness in medical interventions. In summarizing the myriad findings, the review underscores the significance of understanding the differential impacts of radiation approaches on inflammation and immune modulation, offering valuable insights for developing innovative therapeutic interventions that harness the immune system in conjunction with RT.

癌症治疗领域正在发生重大变化,重点是采用先进的放射治疗方法,以最大限度地提高疗效和减少不良反应。放射治疗(RT)被认为是癌症和疾病治疗的关键组成部分,它与炎症和免疫反应之间错综复杂的相互作用引起了近期研究的关注。这种探索揭示了对治疗效果产生重大影响的潜在过程。在此背景下,研究人员探讨了在不同临床情况下将支气管镜检查与 RT 结合使用的潜在优势,以及近距离放射治疗的针对性影响。同时,放射治疗具有多方面的作用,如 DNA 修复、细胞清除和产生称为损伤相关分子模式的免疫应激信号分子,从而阐明了它们在治疗各种疾病方面的有效性。体外射束 RT 利用光子、电子、质子或碳离子等粒子实现了多功能性,每种粒子都具有独特的优势。先进的 RT 技术为不断发展的领域做出了贡献,FLASH、空间分化 RT 等新兴技术将彻底改变这一领域。对 RT 的理解,就是努力提高治疗效果、减少副作用,以及促进癌症和非癌症患者的个性化创新治疗。在取得这些进步之后,我们的目标已定,那就是开创一个新时代,让 RT 成为医疗干预精准有效的基石。在总结众多研究结果的过程中,该综述强调了了解辐射方法对炎症和免疫调节的不同影响的重要性,为开发创新的治疗干预措施提供了宝贵的见解,这些干预措施可在使用 RT 的同时利用免疫系统。
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Cell Biochemistry and Function
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