肝脏疾病微RNA胞间传递研究的现状与思考

微RNA(microRNAs)是一类通过调控基因表达参与机体生理、病理过程的非编码RNA.近来,研究证实其可在肝脏不同类型细胞间进行传递,以调控靶细胞的功能,从而参与肝脏疾病的发生发展.但其在不同类型肝脏细胞间传递的直接实验证据--细胞共培养实验仍需考虑:不同类型细胞在共培养时的数量比例及miRNAs在不同细胞间传递的方向.miRNAs的胞间传递作为肝脏疾病病理机制的重要理论创新,在研究过程中仍需要考虑临床实践.本文对近期关于微RNA胞间传递与肝脏疾病的研究进行综述,以期促进对相关研究的思考.     

茄子microRNAs与其靶基因的生物信息学预测

microRNAs(miRNAs)是一类在真核生物中发现的长度为21 nt左右、非编码、内源性的单链小分子RNA,通过与靶基因的互补发挥转录后水平的负调控作用。目前,已在许多物种中报道了miRNAs的存在,然而还未见关于茄子miRNAs的报道。根据miRNAs在植物中的高度保守性及其前体的二级结构特征,文章通过同源预测的方法,将已知植物的miRNAs与茄子EST数据库比对,经过一系列的筛选,最终预测到12个家族的16条茄子miRNAs,其中包括3个miRNA家族的正义/反义miRNAs,而miR390和miR399家族的正义/反义miRNAs属于第一次发现。文章还通过在线软件psRNATarget预测到15条茄子miRNAs的71个靶基因,这些靶基因主要编码与茄子生长发育、新陈代谢以及胁迫响应等过程相关的蛋白。     

MicroRNAs对斑马鱼发育的调控

microRNAs(miRNAs)是一类长度约为22nt的非编码小RNA,从单细胞到多细胞真核生物中都广泛存在,在进化过程中高度保守,对动物发育、生理功能及病理过程都具有重要调控作用。斑马鱼(Danio rerio)是现代生物学研究中广泛使用的模式动物,以斑马鱼为模型研究miRNAs可以揭示miRNAs在脊椎动物中的功能。文章就miRNAs整体缺失对斑马鱼胚胎发育的影响及一些miRNAs在斑马鱼早期发育过程中的调控机制进行了综述,从而为探索miRNAs在脊椎动物中的功能及鱼类的生产育种提供理论基础。     

MicroRNAs参与调控中枢神经系统的发育及脊髓损伤修复

microRNAs（miRNAs）不仅参与神经系统的生长发育、功能完善,还参与脊髓损伤病理及损伤后修复过程。miRNAs能使中枢神经系统按正确的时序性和空间性顺序进行发育和分化,在维持生物体记忆及生物钟方面起着重要作用。miRNAs异常表达同脊髓损伤病理过程相关。目前,体内及体外实验均已证实,miRNAs不仅能够维持神经干细胞增殖,而且可以促进神经元轴突伸长,从而为脊髓损伤的治疗带来新的治疗策略。     

High-throughput sequence analysis of small RNAs in skotomorphogenic seedlings of Brassica rapa ssp. rapa

Skotomorphogenic development is the process by which seedlings adapt to a stressful dark environment. Such metabolic responses to abiotic stresses in plants are known to be regulated in part by microRNAs (miRNAs); however, little is known about the involvement of miRNAs in the regulation of skotomorphogenesis. To identify miRNAs at the genome-wide level in skotomorphogenic seedlings of turnip (Brassica rapa subsp. rapa), an important worldwide root vegetable, we used Solexa sequencing to sequence a small RNA library from seedlings grown in the dark for 4 days. Deep sequencing showed that the small RNAs (sRNAs) were predominantly 21 to 24 nucleotides long. Specifically, 13,319,035 reads produced 359,531 unique sRNAs including rRNA, tRNA, miRNA, small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), and unannotated sRNAs. Sequence analysis identified 96 conserved miRNAs belonging to 36 miRNA families and 576 novel miRNAs. qRT-PCR confirmed that the miRNAs were expressed during skotomorphogenesis similar to the trends shown by the Solexa sequencing results. A total of 2013 potential targets were predicted, and the targets of BrmiR157, BrmiR159 and BrmiR160 were proved to be regulated by miRNA-guided cleavage. These results show that specific regulatory miRNAs are present in skotomorphogenic seedlings of turnip and may play important roles in growth, development, and response to dark environment.     

Genetic markers for diagnosis and pathogenesis of Alzheimer's disease

Alzheimer's disease (AD) is the most common form of dementia in the elderly and represents an important and increasing clinical challenge in terms of diagnosis and treatment. Mutations in the genes encoding amyloid precursor protein (APP), presenilin 1 (PSEN1) and presenilin 2 (PSEN2) are responsible for early-onset autosomal dominant AD. The ε4 allele of the apolipoprotein E (APOE) gene has been recognized as a major genetic risk factor for the more common, complex, late-onset AD. Fibrillar deposits by phosphorylated tau are also a key pathological feature of AD. The retromer complex also has been reported to late-onset AD. More recently, genome-wide association studies (GWASs) identified putative novel candidate genes associated with late-onset AD. Lastly, several studies showed that circulating microRNAs (miRNAs) in the cerebrospinal fluid (CSF) and blood serum of AD patients can be used as biomarkers in AD diagnosis. This review addresses the advances and challenges in determining genetic and diagnostic markers for complex AD pathogenesis.     

MicroRNA expression in the aging mouse thymus

MicroRNAs (miRNAs) have been implicated in the process of aging in many model organisms, such as Caenorhabditis elegans, and in many organs, such as the mouse lung and human epididymis. However, the role of miRNAs in the thymus tissues of the aging mouse remains unclear. To address this question, we investigated the miRNA expression profiles in the thymuses of 1-, 10- and 19-month-old mice using miRNA array and qRT-PCR assays. A total of 223 mouse miRNAs were screened, and the expression levels of those miRNAs exhibited gradual increases and decreases over the course of thymus aging. Fifty miRNAs in the 10-month-old thymus and 81 miRNAs in the 19-month-old thymus were defined as differentially expressed miRNAs (p < 0.05) in comparison with their levels in the 1-month-old mouse, and approximately one-third of these miRNAs were grouped within 11 miRNA clusters. Each miRNA cluster contained 2 to 5 miRNA genes, and most of the cluster members displayed similar expression patterns, being either increased or decreased. In addition, Ingenuity Pathway Analysis (IPA) software and the IPA database were used to analyze the 12 miRNAs that exhibited significant expression changes, revealing that as many as 15 pathways may be involved. Thus, our current study determined the expression profiles of miRNAs in the mouse thymus during the process of aging. The results suggested that these miRNAs could become meaningful biomarkers for studying thymus aging and that the aging-related alternations in miRNA expression may be involved in the regulation of cell proliferation, apoptosis, development and carcinogenesis/tumorigenesis.     

Myotube-derived exosomal miRNAs downregulate Sirtuin1 in myoblasts during muscle cell differentiation

It has recently been established that exosomes can mediate intercellular cross-talk under normal and pathological conditions through the transfer of specific miRNAs. As muscle cells secrete exosomes, we addressed the question of whether skeletal muscle (SkM) exosomes contained specific miRNAs, and whether they could act as “endocrine signals” during myogenesis. We compared the miRNA repertoires found in exosomes released from C2C12 myoblasts and myotubes and found that 171 and 182 miRNAs were exported into exosomes from myoblasts and myotubes, respectively. Interestingly, some miRNAs were expressed at higher levels in exosomes than in their donor cells and vice versa, indicating a selectivity in the incorporation of miRNAs into exosomes. Moreover miRNAs from C2C12 exosomes were regulated during myogenesis. The predicted target genes of regulated exosomal miRNAs are mainly involved in the control of important signaling pathways for muscle cell differentiation (e.g., Wnt signaling pathway). We demonstrated that exosomes from myotubes can transfer small RNAs (C. elegans miRNAs and siRNA) into myoblasts. Moreover, we present evidence that exosome miRNAs secreted by myotubes are functionally able to silence Sirt1 in myoblasts. As Sirt1 regulates muscle gene expression and differentiation, our results show that myotube–exosome miRNAs could contribute to the commitment of myoblasts in the process of differentiation. Until now, myokines in muscle cell secretome provided a conceptual basis for communication between muscles. Here, we show that miRNA exosomal transfer would be a powerful means by which gene expression is orchestrated to regulate SkM metabolic homeostasis.