CN114606235B - 环状rna sirt5及其在非酒精性脂肪性肝病诊治中的应用 - Google Patents
环状rna sirt5及其在非酒精性脂肪性肝病诊治中的应用 Download PDFInfo
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Abstract
本发明涉及一种环状RNA SIRT5及其在非酒精性脂肪性肝病诊治中的应用。本发明揭示了c i rcRNA SIRT5在NAFLD的发生发展中的作用和机制，为丰富NAFLD发病机理及临床治疗提供了理论依据，有助于NAFLD疾病的分子标志物的开发、临床药物的研发。
Description
技术领域
本发明涉及医药技术领域，特别是涉及一种环状RNA SIRT5及其在非酒精性脂肪性肝病诊治中的应用。
背景技术
随着经济的提高、人民生活水平和寿命的提升及生活习惯的改变，非酒精性脂肪性肝病(Nonalcoholic fatty liver disease，NAFLD)的发病率呈快速上升的趋势，NAFLD未来有望取代慢性病毒性肝炎，成为最主要的肝脏慢性疾病，严重威胁人民群众的生命健康。目前，随着诊断水平的提高及机制研究的深入，人们对脂肪肝的发生、发展有了一定的认识，但目前对于NAFLD的治疗，目前国内外尚无有效的针对性治疗药物或手段应用于临床，因此，厘清脂肪肝的发生机制及找到有效的防治措施，仍然是我们长期面临的重要课题。
NAFLD通常是代谢综合征的组成部分，是代谢综合征在肝脏的表现，由此包括中国学者在内的来自世界22个不同国家的学者们达成共识，建议将NAFLD更名为代谢紊乱相关脂肪性肝病(Metabolic-dysfunction-associated fatty liver disease,MAFLD)。NAFLD涉及广泛的临床症状，从以肝细胞内脂肪堆积为特征的良性肝脏疾病脂肪变性，到以炎症、肝细胞损伤和肝纤维化为特征的非酒精性脂肪性肝炎(Nonalcoholic steatohepatitis,NASH)，NASH可进一步发展为肝硬化和肝细胞癌。虽然目前已经清楚肥胖导致脂肪酸在肝脏的过度堆积是脂肪肝发生的主要病因，然而临床上并非所有肥胖患者都并发有NAFLD，也不是所有的NAFLD的患者都有肥胖，这一现象在亚洲人群中更为显著；另外，当脂肪酸在肝脏聚积后，大约只有20-30％的单纯性脂肪肝的患者会进一步发展为NASH和肝坏死。此外，脂肪变性、炎症和纤维化的发展以及关键临床结果之间的关系在个体之间似乎存在很大的差异。这说明肝脏脂质过度积聚只是表象，那么究竟是什么因素导致了脂质在肝脏的异常沉积？肝脏脂质过度积聚又是如何引起NAFLD的发生发展？这些重要的问题仍有待进一步阐释。
线粒体是细胞内的关键细胞器，不仅为细胞提供能量，还是细胞内自由基生成的重要场所，甚至参与调控细胞凋亡。在生理状态下，细胞内的线粒体处于动态变化中，包括线粒体的形态、结构的变化，线粒体的分裂、再生和融合，以及线粒体自噬等。通过自身的动态平衡，线粒体维持着机体内的代谢稳态变化。在NAFLD的发病过程中，线粒体首先通过平衡NAD+/NADH的氧化还原状态和增加线粒体的延伸来应对肝细胞内过量的脂质超载。随着疾病的进展，线粒体的适应性和灵活性下降，导致活性氧(Reactive oxygen species,ROS)生成增加，进而导致线粒体DNA(Mitochondrial DNA,mtDNA)的氧化损伤，线粒体结构异常(表现为巨型线粒体、线粒体嵴丢失和线粒体颗粒混浊)，脂质过氧化等线粒体代谢稳态失衡现象，恶化疾病进程。由此看出，肝脏线粒体损伤不仅是NAFLD早期始发事件，且随NAFLD进程不断加重，更是贯穿了NAFLD病程的始终，鉴于影响NAFLD发生和进展的脂肪酸β-氧化和氧化应激均主要发生于线粒体，甚至还有学者提出NAFLD是一种线粒体疾病，因此线粒体损伤一直是NAFLD发病机制研究领域的重点。若线粒体的动态变化平衡被破坏，则会影响到线粒体的功能，甚至影响细胞的生存。而线粒体能量代谢障碍是细胞氧化应激的主要原因，表现为ROS的形成。因此，线粒体代谢稳态的失衡可能才是影响肝脏脂质异常积聚及NAFLD发生发展的关键因素。那么又是什么因素导致了线粒体代谢稳态的失衡呢？
环状RNA(Circular RNAs,circRNAs)是一种新近发现的内源性具有闭环结构的非编码RNA，具有稳定性、广泛性、保守性及组织特异性等特点，在基因表达调控中发挥重要作用。circRNAs不受RNA外切酶影响，表达更稳定且不易降解，在真核细胞转录组中广泛表达。研究发现，circRNAs在许多人类疾病，包括癌症、神经退行性改变、心血管疾病等中表达异常。虽然目前已有海量的circRNAs被识别出来，但迄今只有极少数的circRNAs分子的功能得以被揭示，其所涉及的功能主要集中在miRNA海绵、亲本基因的顺式调控、竞争性结合RNA结合蛋白(RNA-binding protein,RBP)和翻译短肽等。
在NAFLD研究领域，由于临床肝脏组织标本获取受限等因素，过去大多数的研究仅局限于利用棕榈酸或油酸干预人肝癌细胞株或正常肝细胞株建立NAFLD的体外模型，或者使用高脂饮食诱导小鼠NAFLD模型，进而通过测序、沉默及过表达的手段进行体外的研究探讨，鲜有涉及NAFLD患者肝组织中circRNAs的表达谱及进而探讨潜在的分子机制和分子靶点的研究报道。最近我们关注到的，2020年中山大学苏士成教授团队发表在CELL上的研究提示，通过提取人肝组织原代成纤维细胞进行基因芯片检测，发现hsa_circ_0089762在NASH肝硬化的肝组织中低表达，并将其命名为脂肪肝炎相关circRNA ATP5B调节因子(Steatohepatitis-associated circRNA ATP5B regulator，SCAR)。进一步的表型和机制研究发现circRNA SCAR可通过直接与mPTP复合体ATP合成酶的ATP5B结合，并在静息状态下阻断CypD与mPTP的相互作用等。这一研究首次关注了NASH伴有肝硬化患者肝星状细胞内circRNAs表达情况，并进一步揭示了人NASH向肝纤维化肝硬化发生发展的重要分子机制。NASH向肝硬化的发展属于NAFLD疾病进展的相对晚期阶段，而关于circRNAs在早期NAFLD患者的作用和机制研究，目前国内外鲜有相关文献报道，也缺乏相应的分子标志物和药物。
发明内容
本发明的目的在于提供一种可用于非酒精性脂肪性肝病的诊断、治疗或病情监测的环状RNA。
本发明提供了一种环状RNA SIRT5，所述环状RNA SIRT5的核苷酸序列如SEQ IDNO.1所示。
本发明还提供了一种DNA片段，其编码所述的环状RNA SIRT5，如SEQ ID NO.2所示。
本发明还提供了一种重组表达载体，所述重组表达载体含有所述的DNA片段。
在一些实施例中，所述重组表达载体为慢病毒载体。
本发明还提供了一种宿主细胞，所述宿主细胞含有所述的重组表达载体。
在一些实施例中，所述宿主细胞为HepG2细胞。
本发明还提供了所述的环状RNA SIRT5、DNA片段、重组表达载体或宿主细胞在制备用于非酒精性脂肪性肝病的诊断、治疗或病情监测的产品中的应用。
在一些实施例中，所述产品为试剂、试剂盒、药物或装置。
本发明还提供了一种用于治疗非酒精性脂肪性肝病的药物，包括所述的环状RNASIRT5、DNA片段、重组表达载体或宿主细胞，以及可药用的辅料。
本发明还提供了所述的环状RNA SIRT5、DNA片段、重组表达载体或宿主细胞在制备用于抑制hsa-miR-150-5P表达或提高SIRT5蛋白表达的产品中的应用。
本发明首先在正常肝脏和NAFLD肝组织样本中证实了线粒体代谢稳态失衡、circRNA SIRT5与NAFLD发生发展的相关性，进而在小鼠NAFLD模型中，观察了circRNASIRT5调控肝细胞内线粒体代谢稳态失衡的动态进程，探讨了SIRT5是否可作为干预NAFLD进程的表观遗传学调控靶点，以及合理调控circRNA SIRT5在肝细胞中的表达是否有助于维持线粒体代谢稳态的平衡，进而改善NAFLD患者预后。本发明揭示了circRNA SIRT5在NAFLD的发生发展中的作用和机制，为丰富NAFLD发病机理及临床治疗提供了理论依据，有助于NAFLD疾病的分子标志物的开发、临床药物的研发。
附图说明
图1为本发明一实施例的circRNA SIRT5参与NAFLD的发生发展调控示意图；
图2为本发明一实施例的NAFLD患者肝脏组织circRNA二代测序及分析验证的结果；其中，A：人肝脏组织HE染色；B：人肝脏组织circRNA二代测序热图；C：维恩分析；D：差异表达circRNAs的表达验证；E：人肝脏组织线粒体ECAR；F：人肝脏组织线粒体OCR；G：环状RNA成环验证。
图3为本发明一实施例的circRNA SIRT5减少NAFLD小鼠肝脏组织中脂肪沉积、ROS及炎症因子释放的结果；其中A：小鼠肝脏组织细胞内及线粒体内ROS表达量检测；B：小鼠血浆中炎性因子的表达量；C：正常小鼠及NAFLD小鼠的体型观；D：小鼠肝脏组织大体观及油红染色观察脂肪变性情况。
图4为本发明一实施例的circRNA SIRT5抑制HepG2细胞脂毒性模型中脂肪沉积和炎症因子释放的结果；其中，A：棕榈酸处理后circRNA表达情况；B：棕榈酸处理后circRNA表达情况；C：棕榈酸处理后细胞中肝酶水平；D：光镜下观察circRNA SIRT5对PA处理hepG2细胞脂肪变性的影响；E：hepG2细胞内及线粒体内ROS表达量检测；F：细胞内SIRT5 mRNA表达水平。
图5为本发明一实施例的circRNA SIRT5潜在作用机制相关性检测与靶点预测结果；其中，A：人肝脏组织中has-miR-150-5P的表达水平；B：小鼠肝脏组织中has-miR-150-5P的表达水平；C：hepG2细胞中has-miR-150-5P的表达水平；D：人肝脏组织中SIRT5的表达水平；E：小鼠肝脏组织中SIRT5的表达水平；F：hepG2细胞中SIRT5的表达水平；G：circRNASIRT5与hsa-miR-150-5P预测结合位点情况。
图6为本发明一实施例的circRNA SIRT5可调控SIRT5及其下游信号通路蛋白表达的结果；其中，A：小鼠肝脏组织中SIRT5、ECH1、VLCAD和SOD1的表达；B：hepG2细胞中SIRT5、ECH1、VLCAD和SOD1的表达；。
具体实施方式
为了更加简洁明了的展示本发明的技术方案、目的和优点，下面结合具体实施例及其附图对本发明做进一步的详细描述。可以理解，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。
本发明通过选取人正常肝组织、轻度NAFLD(大泡性脂肪变5％-10％)、中度NAFLD(大泡性脂肪变30-60％)和重度NAFLD(大泡性脂肪变>60％)的肝脏组织各5例进行circRNAs二代测序，结果提示，与正常肝脏相比，随着病变严重程度的加重，存在有10个与疾病严重程度呈相关性表达的circRNAs，表明这些circRNAs可能在NAFLD的发生发展中扮演着重要角色。其中3个circRNAs表达水平降低，7个表达水平升高。随后对相关数据进行差异表达分析、维恩分析、基因功能分析、信号通路分析及表达验证实验，我们发现novel_circ_0029917在这10个差异基因中表达变化幅度最大，与疾病的严重程度相关性最强。我们进一步对其母本基因进行分析发现，novel_circ_0029917由SIRT5基因的外显子反向剪接而来，为便于理解，我们将novel_circ_0029917命名为circRNA SIRT5。
Sirtuins是一类烟酰胺腺嘌呤二核苷酸(Nicotinamide adenine dinucleotide,NAD+)依赖的蛋白脱酰酶和/或ADP核糖基转移酶，包括7个成员，即SIRT1-SIRT7。不同的Sirtuin成员具有不同的亚细胞定位和功能。SIRT1、6、7主要定位于细胞核内，SIRT3、4、5分布于线粒体内。目前已知，sirtuins可以调节多种生物过程：DNA修复、基因表达、细胞生存、新陈代谢、衰老等。在线粒体sirtuins中，SIRT5对带负电荷的酰基赖氨酸修饰表现出独特的亲和力，并进行蛋白去琥珀酰化、去丙二酰化和去戊二酰化反应。SIRT5在机体内分布广泛，在脑、心、肝、肾、肌肉和睾丸中的含量最高。SIRT5可通过调节葡萄糖氧化、酮体形成、脂肪酸氧化、氨和ROS解毒等生物过程，维持线粒体代谢和细胞内稳态。SIRT5敲除导致小鼠肝脏和肌肉中的β氧化受损及中长链酰基肉碱的积累。SIRT5可直接结合并通过去琥珀酰化激活铜/锌超氧化物歧化酶，进而增强SOD1介导的ROS解毒功能。SIRT5敲低或敲除的细胞显示NADPH和GSH水平降低，导致清除ROS的能力受损，并增加对氧化应激的敏感性。一直以来，Sirtuins家族都是代谢疾病研究领域的热点。在NAFLD领域，研究表明SIRT5可通过调节ob/ob小鼠中代谢相关蛋白去乙酰化减轻肝脏脂肪变性。另外，SIRT5敲除可使高脂喂养的小鼠肝脏线粒体中链脂肪酸氧化能力受损，加重脂肪肝。由此，我们认为SIRT5作为代谢感受器蛋白，在维持线粒体代谢稳态参与NAFLD发生发展中发挥重要作用。另外，我们也发现人肝脏组织中的SIRT5蛋白表达变化与肝细胞脂肪积聚及NAFLD发展进程密切相关。这些研究表明：单纯的肥胖及胰岛素抵抗并不是肝细胞脂肪积聚和NAFLD的决定因素，而SIRT5介导的线粒体代谢稳态的失衡可能在NAFLD的发生和发展中起更为重要的作用。那么SIRT5蛋白表达异常的启动因素和内在动力是什么？由SIRT5反向剪接形成的circRNA SIRT5是否在此过程中扮演了重要角色？
为此，我们初步探讨了circRNA SIRT5对SIRT5介导的线粒体代谢稳态失衡的影响。通过合成circRNA SIRT5过表达慢病毒并转染HepG2细胞。在使用棕榈酸建立体外脂毒性模型中，我们发现过表达circRNA SIRT5可以显著促进长链脂肪酸的β-氧化和抑制线粒体ROS的生成，进而降低炎症因子的释放，缓解细胞的脂毒性。由此我们推测circRNA SIRT5可能通过调节肝细胞线粒体代谢稳态调控NAFLD的发生发展，但其参与调节NAFLD的具体机制尚不明确。我们进一步通过生物信息学分析发现，SIRT5受hsa-miR-150-5p靶向调节，而circRNA SIRT5上正好存在hsa-miR-150-5p的结合位点。我们还发现尽管circRNA SIRT5在人和小鼠之间的同源性较差，但SIRT5和hsa-miR-150-5p在人和小鼠之间序列高度保守，加上此前已有研究报道利用人源非编码RNA干预小鼠疾病进程的先例，为我们后续体内模型验证circRNA SIRT5的表型及功能打下坚实基础。同时，我们还发现，circRNA SIRT5在人NAFLD肝脏组织和HepG2体外脂毒性模型中表达与hsa-miR-150-5p呈负相关，而与SIRT5的表达呈正相关。我们进一步推测circRNA SIRT5可能通过吸附hsa-miR-150-5p，进而上调SIRT5的表达，参与NAFLD的发生发展调控，如图1所示。
综上所述，本发明首先在正常肝脏和NAFLD肝组织样本中证实是否存在线粒体代谢稳态失衡、circRNA SIRT5与NAFLD发生发展的相关性，进而在小鼠NAFLD模型中，观察circRNA SIRT5调控肝细胞内线粒体代谢稳态失衡的动态进程，探讨SIRT5是否可作为干预NAFLD进程的表观遗传学调控靶点，以及合理调控circRNA SIRT5在肝细胞中的表达是否有助于维持线粒体代谢稳态的平衡，进而改善NAFLD患者预后。本发明揭示了circRNA SIRT5在NAFLD的发生发展中的作用和机制，为丰富NAFLD发病机理及临床治疗提供了理论依据，有助于NAFLD疾病的分子标志物的开发、临床药物的研发。
circRNA SIRT5序列：
UAAAUGGAAAUGUUUUCUAACAUAUAAAAACCUACAGAAGAAGAAAAUAAUUUUCUGGAUCAAAUUAGAAGUCUGUAUUAUAUUGAUGUCUCCAGAUUCAAAUAUAUUAGAAAGCAGCCGUGGAGACAACCAUCUUCAUUUUGGGAGAAAUAACUAAAGUAGCUUAUUUAAAACUCGAUGUACCUCUUGUGGAGUUGUGGCUGAGAAUUACAAGAGUCCAAUUUGUCCAGCUUUAUCAGGAAAAGGGCUCCAGAACCUGGAACUCAAGAUGCCAGCAUCCCAGUUGAGAAACUUCCCCG
circRNA SIRT5 cDNA基因序列：
TAAATGGAAATGTTTTCTAACATATAAAAACCTACAGAAGAAGAAAATAATTTTCTGGATCAAATTAGAAGTCTGTATTATATTGATGTCTCCAGATTCAAATATATTAGAAAGCAGCCGTGGAGACAACCATCTTCATTTTGGGAGAAATAACTAAAGTAGCTTATTTAAAACTCGATGTACCTCTTGTGGAGTTGTGGCTGAGAATTACAAGAGTCCAATTTGTCCAGCTTTATCAGGAAAAGGGCTCCAGAACCTGGAACTCAAGATGCCAGCATCCCAGTTGAGAAACTTCCCCG
具体研究方法如下：
1.分析circRNA SIRT5、hsa-miR-150-5p及线粒体代谢稳态相关蛋白在NAFLD患者及健康人肝脏组织中的相关性
a.生化指标的测定：外周血或肝脏组织中细胞因子(IL-1β、TNF-α、IL-6、等)反映炎症激活状态；检测肝功能指标(ALT、AST)；
b.NAFLD相关指标的检测：利用油红O、HE染色观察肝脏脂质聚集及形态学改变；利用qRT-PCR、Western Blotting或免疫组化方法检测检测肝组织炎症介质(TNF-α、IL-1、IL-6)mRNA及蛋白水平；
c.线粒体代谢稳态相关指标检测：Seahorse检测肝脏线粒体的代谢表型；WesternBlotting检测SOD1、ECH1、VLCAD、SIRT5；DCFH-DA法和mitoSOX法分别检测细胞质和线粒体中的ROS；Seahorse检测肝细胞线粒体能量代谢变化；
d.circRNA SIRT5在NAFLD患者肝脏组织中的表达及定位：qRT-PCR等方法检测circRNA SIRT5的表达；RnaseR消化耐受实验验证circRNA SIRT5的环化情况；并结合上述指标分析circRNA SIRT5、hsa-miR-150-5p及线粒体代谢稳态相关指标三者的相关性；
2.验证circRNA SIRT5在NAFLD小鼠模型中的作用
a.动物模型构建：采用高脂饮食(High-fat diet,HFD)喂养野生型C57BL/6J小鼠建立NAFLD小鼠模型，并CD喂养的野生型C57BL/6J小鼠作为对照组。AAV8肝脏特异性病毒载体转染建立小鼠circRNA SIRT5过表达模型；
b.生化检测：于实验前、实验开始后1，8周、16周抽取动物血液。通过检测血清细胞因子(IL-1β、TNFα、IL-6等)反映实验动物的炎症激活状态；检测肝功能指标(ALT、AST)；
c.脂质聚积的形态学:用油红O染色法在冰冻切片上检测肝脏脂肪变性程度；
d.线粒体代谢稳态相关指标检测：Western Blotting检测SOD1、ECH1、VLCAD、SIRT5；DCFH-DA法和mitoSOX法分别检测细胞质和线粒体中的ROS；
e.circRNA SIRT5在NAFLD小鼠肝脏组织中的表达及定位：qRT-PCR检测circRNASIRT5、hsa-miR-150-5p及线粒体代谢稳态相关指标表达水平，并结合上述指标分析三者之间的相关性；
3.细胞水平上验证circRNA SIRT5在NAFLD肝细胞体外模型中的作用
a.NAFLD体外模型构建：采用棕榈酸诱导HepG2肝细胞株脂毒性模型，circRNASIRT5过表达慢病毒转染细胞，以期明确circRNA SIRT5在NAFLD体外模型中的作用；
b.生化指标的测定：上清及细胞匀浆中肝功能指标及细胞因子(IL-1β、TNFα、IL-6等)；
d.脂质聚积的形态学:用油红O染色法检测细胞脂肪变性程度；
e.线粒体代谢稳态相关指标检测：Western Blotting检测SOD1、ECH1、VLCAD、SIRT5；
f.circRNA SIRT5在NAFLD体外模型中的表达及鉴定：qRT-PCR检测circRNA SIRT5表达水平，并结合上述指标分析circRNA SIRT5、hsa-miR-150-5p及线粒体代谢稳态相关指标三者的相关性；
关键技术说明：
Seahorse检测：为观察线粒体功能改变与NAFLD的联系，我们将对NAFLD患者和正常人肝脏组织样本进行Seahorse检测。Seahorse XFp分析仪是进行体外和其它有限量样品中代谢表型常规测试的理想工具，目前已广泛应用于基础实验研究。我们首先采用蔗糖法进行线体提纯，这些化合物(寡霉素、FCCP以及鱼藤酮和抗霉素a的混合物)分别被连续注射以测量ATP产生、最大呼吸和非线粒体呼吸。然后利用这些参数和基础呼吸计算质子漏和基础呼吸容量。
实验结果：
1.对健康人、NAFLD(大泡性脂肪肝5-10％)、NAFLD(大泡性脂肪肝30-60％)及NAFLD(大泡性脂肪肝>60％)的肝脏组织标本各5例进行circRNAs测序，得到几百个潜在的致病circRNAs。取各组NAFLD分别与健康人进行差异分析，然后进行维恩分析寻找潜在共性的致病circRNAs，结果提示，找到10个共性差异circRNAs，其中3个低表达，7个高表达。随后对这10个circRNAs进行基因功能分析、信号通路分析及表达验证实验，我们发现发现novel_circ_0029917在这10个差异基因中变化幅度最大，表达变化与疾病的严重程度相关性最强。我们进一步使用RNase R消化实验鉴定circRNA SIRT5环化性质，结果提示circRNASIRT5呈环形，且耐受RNase R酶消化，见图2。
2.构建高脂饮食喂养的野生型C57BL/6J小鼠NAFLD模型，使用circRNA SIRT5过表达慢病毒转染后，结果提示，circRNA SIRT5可显著缓解小鼠肝脏组织的脂肪病变，减少mROS和cROS的表达，同时还可以抑制NAFLD引起的肝细胞炎症因子释放，提示circRNASIRT5可纠正NAFLD引起的线粒体代谢稳态失衡，见图3。
3.我们合成circRNA SIRT5过表达慢病毒并转染HepG2细胞。在使用棕榈酸建立体外脂毒性模型中，我们发现过表达circRNA SIRT5可以显著促进长链脂肪酸的β-氧化和抑制线粒体ROS的生成，进而降低炎症因子的释放，缓解细胞的脂毒性，见图4。
4.为探讨circRNA SIRT5发挥作用的潜在机制，我们通过使用CPC、CNCI和PFAM三个常见的蛋白潜能预测软件对其是否具有蛋白编码潜能进行分析，结果提示，circRNASIRT5的IRES分数为0.728266，均不具有蛋白翻译潜能。进一步我们使用miRDB、TargetScan及RNA Hydrid等预测软件，进行circRNA/miR/mRNA之间相互结合的潜在位点进行分析，发现circRNA SIRT5存在有3个可与hsa-miR-150-5p结合的位点，且hsa-miR-150-5p可靶向结合SIRT5 mRNA。随后我们在人和小鼠NAFLD肝脏组织及HepG2细胞脂毒性模型中发现hsa-miR-150-5P的表达升高，而SIRT5的mRNA表达降低。在circRNA SIRT5过表达的HepG2细胞脂毒性模型中，circRNA SIRT5可显著抑制hsa-miR-150-5P的表达，同时可以解除hsa-miR-150-5P对SIRT5 mRNA的抑制作用，见图5。
5.随后，我们在circRNA SIRT5过表达的小鼠NAFLD模型和HepG2细胞脂毒性模型中观察circRNA SIRT5对SIRT5蛋白及线粒体代谢稳态相关指标的影响。结果提示，circRNASIRT5可上调小鼠肝脏和HepG2细胞中的SIRT5蛋白表达，进而上调信号通路下游线粒体代谢稳态相关蛋白(ECH1、VLCAD及SOD1)的表达，见图6。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。
序列表
<110> 四川大学华西医院
<120> 环状RNA SIRT5及其在非酒精性脂肪肝诊治中的应用
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 299
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 1
uaaauggaaa uguuuucuaa cauauaaaaa ccuacagaag aagaaaauaa uuuucuggau 60
caaauuagaa gucuguauua uauugauguc uccagauuca aauauauuag aaagcagccg 120
uggagacaac caucuucauu uugggagaaa uaacuaaagu agcuuauuua aaacucgaug 180
uaccucuugu ggaguugugg cugagaauua caagagucca auuuguccag cuuuaucagg 240
aaaagggcuc cagaaccugg aacucaagau gccagcaucc caguugagaa acuuccccg 299
<210> 2
<211> 299
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
taaatggaaa tgttttctaa catataaaaa cctacagaag aagaaaataa ttttctggat 60
caaattagaa gtctgtatta tattgatgtc tccagattca aatatattag aaagcagccg 120
tggagacaac catcttcatt ttgggagaaa taactaaagt agcttattta aaactcgatg 180
tacctcttgt ggagttgtgg ctgagaatta caagagtcca atttgtccag ctttatcagg 240
aaaagggctc cagaacctgg aactcaagat gccagcatcc cagttgagaa acttccccg 299
Claims (10)
1.一种环状RNA SIRT5，其特征在于，所述环状RNA SIRT5的核苷酸序列如SEQ ID NO.1所示。
2.一种DNA片段，其特征在于，其编码权利要求1所述的环状RNA SIRT5。
3.一种重组表达载体，其特征在于，所述重组表达载体含有权利要求2所述的DNA片段。
4.根据权利要求3所述的重组表达载体，其特征在于，所述重组表达载体为慢病毒载体。
5.一种宿主细胞，其特征在于，所述宿主细胞含有权利要求3或4所述的重组表达载体。
6.根据权利要求4所述的宿主细胞，其特征在于，所述宿主细胞为HepG2细胞。
7.权利要求1所述的环状RNA SIRT5、权利要求2所述的DNA片段、权利要求3～4任一项所述的重组表达载体或权利要求5～6任一项所述的宿主细胞在制备用于非酒精性脂肪性肝病的诊断、治疗或病情监测的产品中的应用。
8.根据权利要求7所述的应用，其特征在于，所述产品为试剂、试剂盒、药物或装置。
9.一种用于治疗非酒精性脂肪性肝病的药物，其特征在于，包括权利要求1所述的环状RNA SIRT5、权利要求2所述的DNA片段、权利要求3～4任一项所述的重组表达载体或权利要求5～6任一项所述的宿主细胞，以及可药用的辅料。
10.权利要求1所述的环状RNA SIRT5、权利要求2所述的DNA片段、权利要求3～4任一项所述的重组表达载体或权利要求5～6任一项所述的宿主细胞在制备用于抑制hsa-miR-150-5P表达或提高SIRT5蛋白表达的产品中的应用。
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Citations (3)
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| CN104010651A (zh) * | 2011-09-07 | 2014-08-27 | 康奈尔大学 | 通过靶向Sirt5治疗癌症的方法 |
| CN107988228A (zh) * | 2010-05-03 | 2018-05-04 | 库尔纳公司 | 通过抑制沉默调节蛋白(sirt)的天然反义转录物而治疗沉默调节蛋白(sirt)相关疾病 |
| CA3125511A1 (en) * | 2019-02-08 | 2020-08-13 | Curevac Ag | Coding rna administered into the suprachoroidal space in the treatment of ophthalmic diseases |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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-
2022
- 2022-03-25 CN CN202210305595.6A patent/CN114606235B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988228A (zh) * | 2010-05-03 | 2018-05-04 | 库尔纳公司 | 通过抑制沉默调节蛋白(sirt)的天然反义转录物而治疗沉默调节蛋白(sirt)相关疾病 |
| CN104010651A (zh) * | 2011-09-07 | 2014-08-27 | 康奈尔大学 | 通过靶向Sirt5治疗癌症的方法 |
| CA3125511A1 (en) * | 2019-02-08 | 2020-08-13 | Curevac Ag | Coding rna administered into the suprachoroidal space in the treatment of ophthalmic diseases |
Non-Patent Citations (7)
| Title |
|---|
| Bello JH等.Homo sapiens sirtuin 5 (SIRT5), transcript variant 26, non-coding RNA.《GenBank Database》.2022,Accession No.NR_164856.1. * |
| Bin Shang 等.Circ-Klhl8 overexpression increased the therapeutic effect of EPCs in diabetic wound healing via the miR-212-3p/SIRT5 axis.《Journal of diabetes and its complications》.2021,全文. * |
| Daofeng Zheng等.SIRT5 alleviates hepatic ischemia and reperfusion injury by diminishing oxidative stress and inflammation via elevating SOD1 and IDH2 expression.《Experimental cell research》.2022,全文. * |
| Rybak等.hsa_circ_0130765.《circBase》.2015,chr6:13579003. * |
| Xiao-Ou Zhang等.hsa_circ_17421.《circRNADb》.2014,chr6:13579682-13584457. * |
| 刘旭庆等.环状RNA的产生、研究方法及功能.《遗传》.2019,第469-485页. * |
| 曹山虎 ; 何佳丽 ; 董丽华 ; .M2型丙酮酸激酶的功能及调节.中国细胞生物学学报.2020,(08),第122-129页. * |
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