氧化应激反应性miRNA靶向调控Nrf2-ARE信号通路在眼底疾病发生发展中的作用

核因子E2相关因子2-抗氧化反应原件复合体(nuclear factor erythroid 2 related factor 2,Nrf2-antioxidant response element,ARE)是一种重要的内源性抗氧化应激通路,能够调节细胞内众多抗氧化物的表达,具有维持细胞氧化-抗氧化平衡、抑制凋亡、抗炎等多种生物活性。微小RNA(microRNA,miRNA)是真核生物中广泛存在的能调节基因表达的一类非编码、极短小RNA分子。多种氧化应激反应性miRNA可直接或间接作用于Nrf2-ARE信号通路并调控相关基因的表达,促进或抑制视网膜氧化损伤及眼底退行性变等疾病的发生发展。(国际眼科纵览,2020,44:443-447)     

非编码RNA对葡萄膜黑色素瘤发生机制影响研究新进展

葡萄膜黑色素瘤是成人最常见的一种眼内原发性肿瘤,恶性程度高,患者长期存活率低,易于肝转移。MicroRNA（miRNA）是一种单链非编码微小RNA,参与调节信使RNA的转录后翻译。长链非编码RNA（lncRNA）是一种长链（长度大于200 nt）核糖核酸,不具有蛋白质编码的功能。有研究表明,miRNAs和lncRNAs在葡萄膜黑色素瘤的发生发展过程中均起到调控作用。本文总结了近五年关于miRNAs和lncRNAs对葡萄膜黑色素瘤发生机制影响研究新进展,以期发现更多的可用于该病诊疗的新靶点。     

非编码RNA与白内障发病关系的研究进展

白内障是我国乃至全世界首要致盲眼病,其发病机制仍未明确,也尚无有效药物治疗方式。非编码 RNA(non-coding RNA,ncRNA)是一类不具备蛋白编码功能的RNA。 已有研究证明ncRNA与人类疾病的发生发展密切相关,其中ncRNA与眼科疾病的研究也日益增多。本文主要就微小RNA、 长链非编码RNA和环状RNA在白内障中的研究进展进行总结,发现对微小RNA与白内障关系的研究较多,而长链非编码RNA和环状RNA在白内障发病机制中的研究近2年才逐渐开展,因此需要眼科医生进一步深入探究,从而为白内障的防治提供新思路与新方法。     

Circulating microRNA 122 in the methionine and choline‐deficient mouse model of non‐alcoholic steatohepatitis

Non‐alcoholic steatohepatitis (NASH) is a progressive form of non‐alcoholic fatty liver disease (NAFLD) and is a major cause of liver cirrhosis and hepatic failure. The methionine choline‐deficient diet (MCD) is a frequently used hepatotoxicity animal model of NASH that induces hepatic transaminase (ALT, AST) elevations and hepatobiliary histological changes similar to those observed in human NASH. Liver‐specific microRNA‐122 (miR‐122) has been shown as a key regulator of cholesterol and fatty acid metabolism in adult liver, and has recently been proposed as a sensitive and specific circulating biomarker of hepatic injury. The purpose of this study was to assess miR‐122 serum levels in mice receiving an MCD diet for 0, 3, 7, 14, 28 and 56 days and compare the performance vs. routine clinical chemistry when benchmarked against the histopathological liver findings. MiR‐122 levels were quantified in serum using RT‐qPCR. Both miR‐122 and ALT/AST levels were significantly elevated in serum at all timepoints. MiR‐122 levels increased on average by 40‐fold after 3 days of initiating the MCD diet, whereas ALT and AST changes were 4.8‐ and 3.3‐fold, respectively. In general, miR‐122 levels remained elevated across all time points, whereas the ALT/AST increases were less robust but correlated with the progressive severity of NASH as assessed by histopathology. In conclusion, serum levels of miR‐122 can potentially be used as a sensitive biomarker for the early detection of hepatotoxicity and can aid in monitoring the extent of NAFLD‐associated liver injury in mouse efficacy models. Copyright © 2013 John Wiley & Sons, Ltd.     

Using microRNA profiles to predict and evaluate hepatic carcinogenic potential

Novel chemical entities have to be assessed for potential adverse effects in exposed human populations, including increased cancer incidence. The liver is an organ of particular interest for such evaluations, due to its central metabolic and detoxifying functions that render it a frequent target of exogenous carcinogens. In recent years a number of studies have investigated the use of microRNA (miRNA) biomarkers to facilitate the identification, characterization, and mechanistic understanding of chemical hepatocarcinogens. In this review we discuss the main findings of these studies, the potential biological significance of observed miRNA perturbations, and avenues of future research.     

Detection of gestational diabetes mellitus by circulating plasma and serum microRNAs: A systematic review and meta-analysis

Background and aimsThe diagnostic performance of microRNAs (miRNAs), which have recently emerged as a potential early diagnostic tool capable of detecting gestational diabetes mellitus (GDM) in its early stages, has yet to be systematically investigated. This meta-analysis aims to investigate the diagnostic utility of circulating plasma or serum miRNAs in detecting GDM patients.MethodsEligible studies were included and assessed for risk of bias with the Quality Assessment of Diagnostic Accuracy Studies 2 tool. A bivariate meta-analysis using the hierarchical summary receiver operating characteristic model was performed to estimate the pooled diagnostic value of miRNAs.ResultsTwelve studies (32 index tests) cumulating a total of 1768 patients were included in the present study. The pooled sensitivity of miRNAs was 74.5% (95% confidence interval [CI]: 63.7–82.9) and the pooled specificity was 84.1% (95% CI: 76.8–89.3). The overall area under the curve was 0.869 (95% CI: 0.818–0.907) with a relatively narrow 95% confidence region and a wide 95% prediction region.ConclusionmiRNAs may emerge as a promising diagnostic tool in detecting GDM. Further cross-sectional cohort studies with larger sample sizes and more heterogeneous populations, and studies evaluating the accuracy of multiple miRNAs in diagnosing GDM are required to confirm our findings.     

PrLZ基因3′-UTR区荧光素酶报告基因载体的构建及活性检测

目的分析前列腺亮氨酸拉链(PrLZ)基因3′非编码区(3′-UTR)可能的微小RNA(miRNA)调控位点,构建PrLZ基因3′-UTR荧光素酶报告载体,为研究PrLZ基因转录后的miRNA调控提供有效的工具。方法使用PCR方法从PrLZ阳性的C4-2细胞中扩增含PrLZ基因3′-UTR区序列,插入到T-Vector载体上,提高PCR产物的连接、克隆效率,通过Xbal和BamHI限制性内切酶双酶切后将其连入经相同内切酶双酶切后的荧光素酶报告基因载体pLUC中,构建pLUC-PrLZ 3′-UTR载体,使用生物信息学方法预测PrLZ基因3′-UTR可能是miR-34a的作用靶点,使用慢病毒载体构建对照载体(质粒名称-NC)和过表达miR-34a(质粒名称-miR-34a),包装成病毒颗粒后分别感染293-T细胞,然后通过X-tremeGENE HP DNA Transfection Reagent转染试剂将pLUC空质粒和pLUC-PrLZ 3′-UTR重组质粒分别转染293-T细胞,Promega试剂盒测定荧光素酶的活性。结果得到含PrLZ基因3′-UTR序列的荧光素酶报告重组质粒,并用凝胶电泳和基因测序的方法验证了其序列的正确性。在miR-34a高表达组的293-T细胞中,重组质粒组荧光素酶活性比空质粒组低40%,差异具有统计学意义(P0.05)。结论成功构建PrLZ基因3′-UTR荧光素酶报告载体,miR-34a可以显著降低荧光素酶的活性,PrLZ基因3′-UTR上可能有miR-34a的调控作用靶点。