Alcoholic and Non-Alcoholic Beer Modulate Plasma and Macrophage microRNAs Differently in a Pilot Intervention in Humans with Cardiovascular Risk

Beer is a popular beverage and some beneficial effects have been attributed to its moderate consumption. We carried out a pilot study to test if beer and non-alcoholic beer consumption modify the levels of a panel of 53 cardiometabolic microRNAs in plasma and macrophages. Seven non-smoker men aged 30–65 with high cardiovascular risk were recruited for a non-randomised cross-over intervention consisting of the ingestion of 500 mL/day of beer or non-alcoholic beer for 14 days with a 7-day washout period between interventions. Plasma and urine isoxanthohumol were measured to assess compliance with interventions. Monocytes were isolated and differentiated into macrophages, and plasma and macrophage microRNAs were analysed by quantitative real-time PCR. Anthropometric, biochemistry and dietary parameters were also measured. We found an increase in plasma miR-155-5p, miR-328-3p, and miR-92a-3p after beer and a decrease after non-alcoholic beer consumption. Plasma miR-320a-3p levels decreased with both beers. Circulating miR-320a-3p levels correlated with LDL-cholesterol. We found that miR-17-5p, miR-20a-5p, miR-145-5p, miR-26b-5p, and miR-223-3p macrophage levels increased after beer and decreased after non-alcoholic beer consumption. Functional analyses suggested that modulated microRNAs were involved in catabolism, nutrient sensing, Toll-like receptors signalling and inflammation. We concluded that beer and non-alcoholic beer intake modulated differentially plasma and macrophage microRNAs. Specifically, microRNAs related to inflammation increased after beer consumption and decreased after non-alcoholic beer consumption.     

miR-124抑制食管鳞癌细胞TE-1增殖及侵袭的研究

目的 分析微RNA-124(microRNA-124,miR-124)在食管癌组织中的表达情况,并探讨miR-124在食管鳞癌增殖及侵袭中的作用及机制.方法 采用实时荧光定量RT-PCR检测miR-124在18例食管癌组织(Ⅰ期6例,Ⅱ期8例,Ⅲ期4例,Ⅳ期0例)及相应的癌旁组织中的表达情况.在食管癌细胞TE-1中转染miR-124 mimic,采用细胞集落形成实验、细胞计数试剂盒-8(cell counting kit-8,CCK-8)、细胞划痕实验检测miR-124过表达对食管癌细胞增殖及侵袭的影响.通过Targetscan预测细胞周期蛋白依赖性激酶(cyclin-dependent kinases 4,CDK4)为miR-124的直接靶基因,并用荧光素酶报告基因实验及蛋白质印迹法实验进行验证.结果 食管癌组织miR-124的表达水平较癌旁正常组织明显下降(P＜0.01).通过转染miR-124 mimics使TE-1细胞过表达miR-124,过表达miR-124后TE-1细胞增殖及侵袭能力明显下降(P＜0.01).结论 miR-124在食管癌组织中低表达,miR-124可能通过调控CDK4表达来影响食管癌细胞TE-1增殖及侵袭能力.     

胃癌组织中mRNA-375基因高甲基化及其表达的研究

目的:探讨胃癌组织中miRNA-375基因表达与基因甲基化调控的相关性.方法:2011年3月至8月在天津医科大学总医院通过胃镜检查收集90例新鲜组织活检标本,分为2组,胃癌组54例,非癌对照组36例.应用实时荧光定量反转录PCR检测miRNA-375基因表达,甲基化特异性PCR检测miRNA-375基因启动子区CpG岛甲基化.结果:胃癌组miRNA-375基因表达下调,与非癌对照组相比差异有统计学意义(P<0.05);胃癌组和非癌对照组 miRNA-375基因启动子区高甲基化阳性率分别为62.96%(34/54)和22.22%(8/36),差异有统计学意义(χ2=14.405, P<0.05).中高分化胃癌组织中miRNA-375基因表达高于低分化组,差异有统计学意义(t=2.634,P=0.011);miRNA-375基因启动子区甲基化阳性率中高分化组与低分化组分别为44.44%(8/18)和72.22%(26/36),差异有统计学意义(χ2=3.971,P=0.046).结论:癌组织中存在miRNA-375基因异常低表达及启动子区的高甲基化,miRNA-375基因高甲基化可能抑制miRNA-375基因表达,在胃癌发生发展中发挥重要作用.     

hsa-miR-19a-3p下调靶基因SEMA4C抑制胃癌细胞增殖、侵袭能力

目的:探讨SEMA4C在胃癌中的表达以及hsa-miR-19a-3p下调SEMA4C对胃癌细胞增殖、侵袭能力的影响.方法:收集23例胃癌组织和对应癌旁组织标本,采用免疫组化染色、Real-time PCR方法检测SEMA4C的表达,并分析其与临床资料的相关性;检索Targetscan、Miranda等数据库预测调控SEMA4C基因的microRNA,并利用Real-time PCR、双荧光素酶报告系统检测加以证实;通过脂质体转染使人胃癌细胞系BGC823内过表达hsa-miR-19a-3p,Real-time PCR、Western blot检测SEMA4C的表达,通过细胞集落形成实验和Transwell小室研究BGC823细胞的增殖、侵袭能力的变化.结果:胃癌组织中SEMA4C阳性率87.0%,高于癌旁组织(x2=20.30,P<0.000 1),胃癌组织中SEMA4C mRNA的表达高于癌旁组织(t=13.47,P<0.000 1).SEMA4C的表达与胃癌TNM分期(P=0.036 4)、分化程度(P=0.042 7)、淋巴结转移度(P=0.012 8)有关.生物信息学分析hsa-miR-19a-3p、hsa-miR-214-5p、hsa-miR-138-5p可能调控SEMA4C表达,胃癌组织中hsa-miR-19a-3p的表达低于癌旁组织(t=8.233,P<0.000 1),hsa-miR-214-5p(t=0.846 3,P=0.097 6)、hsa-miR-138-5p(t=1.345,P=0.185 7)表达与癌旁组织无明显差异,hsa-miR-19a-3p转染后荧光素酶表达下降至0.46±0.12(F=5.685,P=0.003 2).hsa-miR-19a-3p转染后,SEMA4C mRNA(F=34.39,P<0.000 1)、SEMA4C蛋白(F=67.49,P<0.000 1)表达减少,BGC823细胞克隆数目减少(F=20.77,P<0.000 1),Transwell下室细胞数较少(F=16.37,P=0.000 4).结论:SEMA4C在胃癌中过表达并与肿瘤的发生发展有关,hsa-miR-19a-3p通过下调靶基因SEMA4C抑制胃癌细胞增殖、侵袭能力来发挥其抗肿瘤效应.     

MicroRNA在胆道闭锁中的研究进展

小分子核糖核酸(microRNA,miRNA,miR)是一类约22个核苷酸的单链非编码RNA,在转录后水平调节基因的表达.miRNA广泛存在于人体各个组织器官中,在细胞增殖、分化、凋亡、胚胎发育、信号传导、肿瘤发生及代谢等过程中发挥着重要作用.近年来研究表明,某些miR-NA表达的改变在胆道闭锁的发生发展中起着重要作用,在该过程中体现出极其复杂的调控机制,同时miRNA的研究也为胆道闭锁的早期诊断和治疗带来了新的希望和思路.本文就miRNA在胆道闭锁中的研究进展做一综述.     

miR-320-3p 调控脂肪细胞分化的研究

目的探讨miR-320-3p 对脂肪细胞分化的影响。方法分离并培养小鼠骨髓间充质干细胞（MSCs）,并对其进行脂肪细胞诱导分化3 d,用qRT-PCR 检测miR-320-3p 在成脂分化过程中的表达水平。在骨髓基质细胞系 ST2 中转染miR-320-3p,对其进行成脂方向诱导分化,用油红O 染色和qRT-PCR 检测miR-320-3p 对ST2 成脂分化的影响。结果MSCs 向脂肪细胞分化过程中miR-320-3p 表达增加（P < 0.01）;与转染阴性对照NC mimics 相比,ST2 细胞转染miR-320-3p mimics 后油红O 染色显示脂肪细胞明显增多,脂肪细胞特异性转录因子过氧化物酶体增殖物激活受体γ(PPARγ)、CCAAT 增强子结合蛋白α（C/EBPα）和脂肪细胞脂肪酸结合蛋白4（FABP4）基因表达显著升高,差异有统计学意义（P < 0.05）。结论miR-320-3p 可促进ST2 向脂肪细胞分化。     

miR-155联合超声对分化型甲状腺癌淋巴结转移的诊断价值分析

目的探讨微小RNA-155(miR-155)联合超声对分化型甲状腺癌淋巴结转移的诊断价值。方法选取接受治疗的分化型甲状腺癌患者112例作为甲状腺癌组,另选取同期收治的结节性甲状腺肿患者30例作为良性对照组。采用实时荧光定量PCR(qPCR)检测2组患者血清及组织中miR-155的相对表达量,分析分化型甲状腺癌患者血清及组织中miR-155的表达与临床病理参数的关系。采用受试者工作特征(ROC)曲线分析术前超声以及血清miR-155对分化型甲状腺癌淋巴结转移的诊断价值。结果甲状腺癌组的血清及组织中miR-155的相对表达量均明显高于良性对照组(P<0.01),有淋巴结转移、肿瘤直径>2 cm、TNM分期Ⅲ+Ⅳ期的患者血清及组织中miR-155的相对表达量分别高于无淋巴结转移、肿瘤直径≤2 cm、TNM分期Ⅰ+Ⅱ期的患者(均P<0.05)。ROC结果显示,超声以及血清miR-155对分化型甲状腺癌淋巴结转移均有一定的诊断价值,曲线下面积分别为:0.839(95%CI:0.760～0.919)、0.837(95%CI:0.763～0.912),但漏诊率和误诊率均超过10%。...     

MicroRNAs in laser-induced choroidal neovascularization in mice and rats: their expression and potential therapeutic targets

Choroidal neovascularization characterizes wet age-related macular degeneration. Choroidal neovascularization formation involves a primarily angiogenic process that is combined with both inflammation and proteolysis. A primary cause of choroidal neovascularization pathogenesis is alterations in pro-and anti-angiogenic factors derived from the retinal pigment epithelium, with vascular endothelium growth factor being mainly responsible for both clinical and experimental choroidal neovascularization. MicroRNAs(miRNAs) which are short, non-coding, endogenous RNA molecules have a major role in regulating various pathological processes, including inflammation and angiogenesis. A review of recent studies with the mouse laser-induced choroidal neovascularization model has shown alterations in miRNA expression in choroidal neovascularization tissues and could be potential therapeutic targets for wet age-related macular degeneration. Upregulation of miR-505(days 1 and 3 post-laser), miR-155(day 14) occurred in retina; miR-342-5 p(days 3 and 7), miR-126-3 p(day 14) in choroid; miR-23 a, miR-24, miR-27 a(day 7) in retina/choroid; miR-505(days 1 and 3) in retinal pigment epithelium/choroid; downregulation of miR-155(days 1 and 3), miR-29 a, miR-29 b, miR-29 c(day 5), miR-93(day 14), miR-126(day 14) occurred in retinal pigment epithelium/choroid. Therapies using miRNA mimics or inhibitors were found to decrease choroidal neovascularization lesions. Choroidal neovascularization development was reduced by overexpression of miR-155, miR-188-5 p, miR-(5,B,7), miR-126-3 p, miR-342-5 p, miR-93, miR-126, miR-195 a-3 p, miR-24, miR-21, miR-31, miR-150, and miR-184, or suppression of miR-505, miR-126-3 p, miR-155, and miR-23/27. Further studies are warranted to determine miRNA expression in mouse laser-induced choroidal neovascularization models in order to validate and extend the reported findings. Important experimental variables need to be standardized; these include the strain and age of animals, gender, number and position of laser burns to the eye, laser parameters to induce choroidal neovascularization lesions including wavelength, power, spot size, and duration.     

HSV-1编码的microRNA在潜伏感染中抗凋亡作用研究进展

在已发现的病毒microRNA中,大多数为疱疹病毒家族所编码,其中HSV-1在潜伏感染过程中编码的miR-LAT通过与靶目标SMAD3与TGF-βmRNA分子的3'端非编码区域(3'-un-translated regions,3'-UTR)互补匹配,并在后转录水平负调控基因的表达,抑制细胞凋亡.本文对HSV-1的miRNAs的抗凋亡作用及其机制进行综述.     

稳定性心绞痛患者血糖水平与循环微小RNA-92a表达关系的研究

目的探讨稳定性心绞痛(SAP)患者循环微小RNA-92a(microRNA-92a,miR-92a)表达与血糖的关系。方法将93例SAP患者分为A组:降糖治疗+空腹血糖(FPG)<7.0 mmol/L 18例,B组:降糖治疗+FPG≥7.0 mmol/L8例,C组:非降糖治疗+FPG<7.0 mmol/L 61例,D组:非降糖治疗+FPG≥7.0 mmol/L 6例。比较4组患者血糖水平与循环miR-92a表达的关系。结果 SAP患者合并2型糖尿病发病率为34.4%,降糖治疗后,仍有30.8%FPG≥7.0 mmol/L。A组与B组miR-92a是否>-0.1或>0.5水平的OR值分别为2.67或0.17,C组与D组OR值分别为0.36、0.48。A组miR-92a>-0.1的患者为88.9%,显著高于C组的41.0%(P<0.01)。B组miR-92a>0.5的患者为75.0%,高于A组的33.3%(P>0.05)。结论单纯测定FPG诊断2型糖尿病漏诊率高。循环miR-92a有助于FPG<7.0 mmol/L的SAP合并2型糖尿病的诊断及疗效评价。