RNA-seq analysis

  • 文章类型: Journal Article
    普通豆(PhaseolusvulgarisL.)是世界范围内种植的经济上重要的粮食作物;但是,它的生产受到各种环境压力的影响,包括寒冷,热,和干旱胁迫。植物U-box(PUB)蛋白家族参与各种生物过程和胁迫反应,但其成员在普通豆中的基因功能和表达模式仍不清楚。这里,我们系统地鉴定了63个U-box基因,包括8个串联基因和55个非串联基因,在普通的豆子里。这些PvPUB基因在11条染色体上分布不均,2号染色体拥有PUB家族的大多数成员,含有10个PUB基因。系统发育树的分析将63个PUB基因分为三组。此外,基于耐寒和冷敏感品种的转录组分析确定了4个差异表达的PvPUB基因,表明它们在耐寒性中的作用。一起来看,这项研究为探索普通豆U-box基因家族的功能方面提供了宝贵的资源,并为新的耐冷普通豆品种的开发提供了重要的理论支持。
    The common bean (Phaseolus vulgaris L.) is an economically important food crop grown worldwide; however, its production is affected by various environmental stresses, including cold, heat, and drought stress. The plant U-box (PUB) protein family participates in various biological processes and stress responses, but the gene function and expression patterns of its members in the common bean remain unclear. Here, we systematically identified 63 U-box genes, including 8 tandem genes and 55 non-tandem genes, in the common bean. These PvPUB genes were unevenly distributed across 11 chromosomes, with chromosome 2 holding the most members of the PUB family, containing 10 PUB genes. The analysis of the phylogenetic tree classified the 63 PUB genes into three groups. Moreover, transcriptome analysis based on cold-tolerant and cold-sensitive varieties identified 4 differentially expressed PvPUB genes, suggesting their roles in cold tolerance. Taken together, this study serves as a valuable resource for exploring the functional aspects of the common bean U-box gene family and offers crucial theoretical support for the development of new cold-tolerant common bean varieties.
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  • 文章类型: Journal Article
    背景:转座因子(TE)是真核生物基因组的主要组成部分。大量的证据表明,尽管它们曾经被认为是“基因组寄生虫”,转座子及其转录本执行特定功能,如早期胚胎发育的调节。了解TEs在吸虫等寄生虫中的作用变得至关重要。肝片吸虫,一种影响人类和牲畜的寄生虫,在不同的环境和主机中经历复杂的生命周期,到目前为止,关于其生命周期调节的知识还很少。
    方法:我们总结了有关肝肝菌中重复元件的数据,并在其生命周期阶段进行了大量RNA-seq分析。TE表达谱进行了分析,关注与先前描述的长链非编码RNA(lncRNAs)的差异表达和潜在同源性。
    结果:差异表达分析揭示了阶段特异性TE转录模式,尤其是在卵和囊虫阶段达到峰值。一些TE与已知的lncRNAs具有同源性,并含有推定的转录因子结合位点。有趣的是,与成虫相比,卵和尾虫中的TE转录水平最高,提示吸虫生命周期转变中的调节作用。
    结论:这些研究结果表明,TEs可能在调节吸虫生命周期转变中发挥作用。此外,TE与lncRNAs的同源性强调了它们在基因调控中的重要性。
    BACKGROUND: Transposable elements (TEs) are major components of eukaryotic genomes. The extensive body of evidence suggests that although they were once considered \"genomic parasites\", transposons and their transcripts perform specific functions, such as regulation of early embryo development. Understanding the role of TEs in such parasites as trematodes is becoming critically important. Fasciola hepatica, a parasite affecting humans and livestock, undergoes a complex life cycle in diverse environments and hosts, and knowledge about its life cycle regulation is scarce so far.
    METHODS: We summarized the data regarding the repetitive elements in F. hepatica and conducted bulk RNA-seq analysis across its life cycle stages. TE expression profiles were analyzed, focusing on differential expression and potential homology with previously described long non-coding RNAs (lncRNAs).
    RESULTS: Differential expression analysis revealed stage-specific TE transcription patterns, notably peaking during egg and metacercariae stages. Some TEs showed homology with known lncRNAs and contained putative transcription factor binding sites. Interestingly, TE transcription levels were highest in eggs and metacercariae compared to adults, suggesting regulatory roles in trematode life cycle transitions.
    CONCLUSIONS: These findings suggest that TEs may play roles in regulating trematode life cycle transitions. Moreover, TE homology with lncRNAs underscores their significance in gene regulation.
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  • 文章类型: Journal Article
    盐度引起的过度活性氧胁迫对甘草的生长构成重大威胁。为了适应盐压力,G.uralensis参与可变剪接(AS)以产生各种蛋白质,帮助其承受盐胁迫的影响。虽然一些研究已经调查了可变剪接对植物胁迫反应的影响,AS与转录调节相互作用以调节G.uralensis的盐应激反应的机制仍然知之甚少。在这项研究中,我们利用高通量RNA测序数据对盐胁迫下的uralensis中不同时间点的AS事件进行了综合分析,外显子跳跃(SE)是主要的AS类型。对不同剪接基因(DSG)进行KEGG富集分析,与AS相关的通路显著丰富,包括RNA转运,mRNA监测,和剪接体。这表明基因的剪接调控,导致盐胁迫条件下的AS事件。此外,植物对盐胁迫的反应途径也得到了丰富,如丝裂原活化蛋白激酶信号通路-植物,类黄酮生物合成,和氧化磷酸化。我们通过AS和qRT-PCR分析重点研究了MAPK途径中的四个差异显著基因。MPK4和SnRK2的选择性剪接类型被跳过外显子(SE)。ETR2和RbohD保留了内含子(RI)和5'选择性剪接位点(A5SS),分别。这四个基因的isoform1的表达水平在不同的组织部位和盐胁迫处理时间显示出不同但显着的增加。这些发现表明,uralensis中的MPK4,SnRK2,ETR2和RbohD激活了isoform1的表达,导致产生更多的isoform1蛋白,从而增强了对盐胁迫的抵抗力。这些发现表明,盐响应性AS直接和间接地控制着乌拉尔草的盐响应。对非生物胁迫期间AS功能和机制的进一步研究可能为增强植物胁迫耐受性提供新的参考。
    Excessive reactive oxygen species stress due to salinity poses a significant threat to the growth of Glycyrrhiza uralensis Fisch. To adapt to salt stress, G. uralensis engages in alternative splicing (AS) to generate a variety of proteins that help it withstand the effects of salt stress. While several studies have investigated the impact of alternative splicing on plants stress responses, the mechanisms by which AS interacts with transcriptional regulation to modulate the salt stress response in G. uralensis remain poorly understood. In this study, we utilized high-throughput RNA sequencing data to perform a comprehensive analysis of AS events at various time points in G. uralensis under salt stress, with exon skipping (SE) being the predominant AS type. KEGG enrichment analysis was performed on the different splicing genes (DSG), and pathways associated with AS were significantly enriched, including RNA transport, mRNA surveillance, and spliceosome. This indicated splicing regulation of genes, resulting in AS events under salt stress conditions. Moreover, plant response to salt stress pathways were also enriched, such as mitogen-activated protein kinase signaling pathway - plant, flavonoid biosynthesis, and oxidative phosphorylation. We focused on four differentially significant genes in the MAPK pathway by AS and qRT-PCR analysis. The alternative splicing type of MPK4 and SnRK2 was skipped exon (SE). ETR2 and RbohD were retained intron (RI) and alternative 5\'splice site (A5SS), respectively. The expression levels of isoform1 of these four genes displayed different but significant increases in different tissue sites and salt stress treatment times. These findings suggest that MPK4, SnRK2, ETR2, and RbohD in G. uralensis activate the expression of isoform1, leading to the production of more isoform1 protein and thereby enhancing resistance to salt stress. These findings suggest that salt-responsive AS directly and indirectly governs G. uralensis salt response. Further investigations into AS function and mechanism during abiotic stresses may offer novel references for bolstering plant stress tolerance.
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  • 文章类型: Journal Article
    几项研究比较了亨廷顿氏病(HD)基因阳性和神经系统正常个体的各个大脑区域的转录组,以确定可能是HD的药物或预后靶标的潜在差异表达基因(DEG)。尽管坚持最佳RNA-Seq分析的技术建议,在这些研究中鉴定为上调的基因中,尚未证明作为HD的预后或治疗靶标是成功的.早期的研究包括比HD基因阳性组年龄大的神经系统正常个体的样本。考虑到大脑衰老引起的逐渐转录变化,我们认为,利用来自较老对照的样本可能导致DEGs的错误识别。为了验证我们的假设,我们重新分析了这项研究的146个样本,可在SRA数据库上访问,并采用倾向评分匹配(PSM)创建一个“虚拟”对照组,其年龄分布与HD基因阳性组具有统计学可比性。我们的研究强调了在基因差异表达分析中使用75岁以上的神经系统正常个体作为对照的不利影响。导致假阳性和阴性。我们最终证明,使用这些旧的对照会导致DEG的错误识别,不利地影响潜在的药物和预后标志物的发现。这强调了在RNA-Seq分析中考虑对照样品年龄的关键作用,并强调将其纳入评估此类研究的最佳实践中。虽然我们主要关注的是高清,我们的研究结果表明,审慎选择适合年龄的对照样本可以显著改善差异表达分析的最佳实践.
    Several studies have compared the transcriptome across various brain regions in Huntington\'s disease (HD) gene-positive and neurologically normal individuals to identify potential differentially expressed genes (DEGs) that could be pharmaceutical or prognostic targets for HD. Despite adhering to technical recommendations for optimal RNA-Seq analysis, none of the genes identified as upregulated in these studies have yet demonstrated success as prognostic or therapeutic targets for HD. Earlier studies included samples from neurologically normal individuals older than the HD gene-positive group. Considering the gradual transcriptional changes induced by aging in the brain, we posited that utilizing samples from older controls could result in the misidentification of DEGs. To validate our hypothesis, we reanalyzed 146 samples from this study, accessible on the SRA database, and employed Propensity Score Matching (PSM) to create a \"virtual\" control group with a statistically comparable age distribution to the HD gene-positive group. Our study underscores the adverse impact of using neurologically normal individuals over 75 as controls in gene differential expression analysis, resulting in false positives and negatives. We conclusively demonstrate that using such old controls leads to the misidentification of DEGs, detrimentally affecting the discovery of potential pharmaceutical and prognostic markers. This underscores the pivotal role of considering the age of control samples in RNA-Seq analysis and emphasizes its inclusion in evaluating best practices for such investigations. Although our primary focus is HD, our findings suggest that judiciously selecting age-appropriate control samples can significantly improve best practices in differential expression analysis.
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  • 文章类型: Journal Article
    植物生物刺激剂作为化学肥料的可持续替代品已经受到关注。细胞外聚合物质(EPS),在植物生长促进根际细菌(PGPRs)分泌的化合物中,被假定为减轻非生物胁迫。本研究旨在研究纯化EPS对非生物胁迫下水稻的影响,并分析其作用机制。进行了盆栽实验,以阐明在糖存在下接种从PGPR纯化的EPS对水稻生长的影响。由于所有EPS在应激后显示SPAD的改善,路德维吉肠杆菌,没有表现出更高的PGP生物活性,如植物激素的产生,固氮,和磷的溶解,选择进行进一步分析。在用EPS或水处理24小时后从发芽种子的胚提取的RNA用于转录组分析。RNA-seq分析显示,在水稻种子中鉴定出215个差异表达基因(DEG),包括139个上调基因和76个下调基因。基因本体论(GO)富集分析表明,富集的GO术语主要与ROS清除过程有关,解毒途径,和对氧化应激的反应。例如,编码OsAAO5的基因的表达,已知该基因在解毒氧化应激中起作用,EPS处理增加了两倍。此外,EPS的应用提高了SPAD和茎和根的干重90%,14%,27%,分别,在干旱胁迫下,盐胁迫下SPAD增加59%。这表明细菌EPS在非生物胁迫下改善了植物的生长。根据我们的结果,我们认为从路德维吉肠杆菌中纯化的EPSs可用于开发水稻生物刺激剂。
    Plant biostimulants have received attention as sustainable alternatives to chemical fertilizers. Extracellular polymeric substances (EPSs), among the compounds secreted by plant growth-promoting rhizobacteria (PGPRs), are assumed to alleviate abiotic stress. This study aims to investigate the effect of purified EPSs on rice under abiotic stress and analyze their mechanisms. A pot experiment was conducted to elucidate the effects of inoculating EPSs purified from PGPRs that increase biofilm production in the presence of sugar on rice growth in heat-stress conditions. Since all EPSs showed improvement in SPAD after the stress, Enterobacter ludwigii, which was not characterized as showing higher PGP bioactivities such as phytohormone production, nitrogen fixation, and phosphorus solubilization, was selected for further analysis. RNA extracted from the embryos of germinating seeds at 24 h post-treatment with EPSs or water was used for transcriptome analysis. The RNA-seq analysis revealed 215 differentially expressed genes (DEGs) identified in rice seeds, including 139 up-regulated and 76 down-regulated genes. A gene ontology (GO) enrichment analysis showed that the enriched GO terms are mainly associated with the ROS scavenging processes, detoxification pathways, and response to oxidative stress. For example, the expression of the gene encoding OsAAO5, which is known to function in detoxifying oxidative stress, was two times increased by EPS treatment. Moreover, EPS application improved SPAD and dry weights of shoot and root by 90%, 14%, and 27%, respectively, under drought stress and increased SPAD by 59% under salt stress. It indicates that bacterial EPSs improved plant growth under abiotic stresses. Based on our results, we consider that EPSs purified from Enterobacter ludwigii can be used to develop biostimulants for rice.
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  • 文章类型: Journal Article
    小麦的条锈病是由真菌病原体Pucciniastriiformisf。sp。小麦(Pst)。通过干扰易感(S)基因选育耐久小麦品种对小麦条锈病的防治具有重要影响。Mingxian169(MX169)对Pst的所有种族都表现出强烈的条锈病敏感性。然而,尚未阐明小麦品种MX169对Pst的敏感性的分子机制和响应基因。这里,我们利用下一代测序技术分析了Pst接种后24、48和120小时(hpi)的“MX169”和高抗性小麦“Zhong4”的转录组学数据。比较转录组分析揭示了在不同时间点的3,494、2,831和2,700个差异表达基因(DEGs)。我们观察到参与光合作用的DEGs的上调,类黄酮生物合成,丙酮酸代谢,硫胺素代谢,和其他生物过程,表明他们参与了MX169对Pst的反应。还鉴定了编码转录因子的DEGs。我们的研究表明,MX169中与条锈病反应相关的潜在易感基因资源对于理解条锈病易感性的机制和提高小麦对Pst的抗性可能是有价值的。
    目的:我们的研究表明,MX169中与条锈病反应相关的潜在易感基因资源对于理解条锈病敏感性和提高小麦对Pst的抗性具有重要价值。
    Stripe rust of wheat is caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst). Breeding durably resistant wheat varieties by disrupting the susceptibility (S) gene has an important impact on the control of wheat stripe rust. Mingxian169 (MX169) showed strong stripe rust susceptibility to all the races of Pst. However, molecular mechanisms and responsive genes underlying susceptibility of the wheat variety MX169 to Pst have not been elucidated. Here, we utilized next-generation sequencing technology to analyze transcriptomics data of \"MX169\" and high-resistance wheat \"Zhong4\" at 24, 48, and 120 h post-inoculation (hpi) with Pst. Comparative transcriptome analysis revealed 3,494, 2,831, and 2,700 differentially expressed genes (DEGs) at different time points. We observed an upregulation of DEGs involved in photosynthesis, flavonoid biosynthesis, pyruvate metabolism, thiamine metabolism, and other biological processes, suggesting their involvement in MX169\'s response to Pst. DEGs encoding transcription factors were also identified. Our study suggested the potential susceptibility gene resources in MX169 related to stripe rust response could be valuable for understanding the mechanisms involved in stripe rust susceptibility and for improving wheat resistance to Pst.
    OBJECTIVE: Our study suggests the potential susceptibility gene resources in MX169 related to stripe rust response could be valuable for understanding the mechanisms involved in stripe rust susceptibility and for improving wheat resistance to Pst.
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  • 文章类型: Journal Article
    优化植物氮(N)的使用并抑制土壤-作物系统中的氮淋失对维持作物产量和减少环境污染至关重要。本研究旨在确定两个N处理之间的数量性状基因座(QTL)和差异表达基因(DEGs),以列出与番茄品种氮相关对比性状相关的候选基因。我们对在两个氮水平下在温室中生长的遗传多样性核心集合(CC)和多亲本高级世代互交(MAGIC)番茄种群进行了表征,并评估了几个与N相关的性状并绘制了QTL。还通过对MAGIC种群的四个亲本中的果实和叶片进行RNA测序,研究了两种N条件下的转录组反应。在生物量和N相关性状的表型水平上观察到对N输入减少的响应存在显着差异。检测到27(27)个QTL的三个目标性状(叶片N含量,叶片氮平衡指数和叶柄NO3-含量),在低氮和高氮条件下为10和6,分别确定了19个QTL的可塑性性状。在转录组水平,在叶片和果实的两种N条件之间检测到4,752和2,405DEGs,分别,其中叶中3628例(50.6%)和果实中1717例(71.4%)是基因型特异性的。当考虑到所有的基因型时,1,677个DEG在器官或组织之间共享。最后,我们整合了DEGs和QTLs分析,以确定最有希望的候选基因。结果强调了番茄中N稳态的复杂遗传结构,以及可用于育种需要较少N输入的番茄品种的新推定基因。
    Optimising plant nitrogen (N) usage and inhibiting N leaching loss in the soil-crop system is crucial to maintaining crop yield and reducing environmental pollution. This study aimed at identifying quantitative trait loci (QTLs) and differentially expressed genes (DEGs) between two N treatments in order to list candidate genes related to nitrogen-related contrasting traits in tomato varieties. We characterised a genetic diversity core-collection (CC) and a multi-parental advanced generation intercross (MAGIC) tomato population grown in greenhouse under two nitrogen levels and assessed several N-related traits and mapped QTLs. Transcriptome response under the two N conditions was also investigated through RNA sequencing of fruit and leaves in four parents of the MAGIC population. Significant differences in response to N input reduction were observed at the phenotypic level for biomass and N-related traits. Twenty-seven (27) QTLs were detected for three target traits (Leaf N content, leaf Nitrogen Balance Index and petiole NO3- content), ten and six at low and high N condition, respectively; while 19 QTLs were identified for plasticity traits. At the transcriptome level, 4,752 and 2,405 DEGs were detected between the two N conditions in leaves and fruits, respectively, among which 3,628 (50.6%) in leaves and 1,717 (71.4%) in fruit were genotype specific. When considering all the genotypes, 1,677 DEGs were shared between organs or tissues. Finally, we integrated DEGs and QTLs analyses to identify the most promising candidate genes. The results highlighted a complex genetic architecture of N homeostasis in tomato and novel putative genes useful for breeding tomato varieties requiring less N input.
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  • 文章类型: Journal Article
    芝麻(SesamumindicumL.)是一种全球栽培的油料作物,以其历史意义和在热带和亚热带地区的广泛生长而闻名。具有显著的营养和药用属性,芝麻在对抗营养不良癌症方面表现出了有希望的效果,糖尿病,和其他疾病,如心血管疾病。然而,芝麻生产面临来自木炭腐烂等环境威胁的重大挑战,干旱,盐度,和内涝压力,给农民造成经济损失。有关抗逆基因和途径的信息的匮乏加剧了这些挑战。尽管它非常重要,目前没有提供芝麻全面信息的平台,严重阻碍了各种胁迫相关基因的挖掘和芝麻的分子育种。为了解决这个差距,在这里一个自由,网络访问,和用户友好的基因组网络资源(SesameGWR,http://backlin。cabgrid.res.in/sesameGWR/)已经开发出这个平台提供了对差异表达基因的关键见解,转录因子,miRNA,和分子标记,如简单的序列重复,单核苷酸多态性,以及与生物和非生物胁迫相关的插入和缺失。.通过RNA-seq数据分析预测了嵌入该网络资源中的功能基因组学信息和注释。考虑到气候变化的影响以及芝麻的营养和药用重要性,这项研究对于理解应激反应至关重要。SesameGWR将成为开发气候适应型芝麻品种的宝贵工具,从而提高这种古老油料作物的生产力。
    Sesame (Sesamum indicum L.) is a globally cultivated oilseed crop renowned for its historical significance and widespread growth in tropical and subtropical regions. With notable nutritional and medicinal attributes, sesame has shown promising effects in combating malnutrition cancer, diabetes, and other diseases like cardiovascular problems. However, sesame production faces significant challenges from environmental threats such as charcoal rot, drought, salinity, and waterlogging stress, resulting in economic losses for farmers. The scarcity of information on stress-resistance genes and pathways exacerbates these challenges. Despite its immense importance, there is currently no platform available to provide comprehensive information on sesame, which significantly hinders the mining of various stress-associated genes and the molecular breeding of sesame. To address this gap, here a free, web-accessible, and user-friendly genomic web resource (SesameGWR, http://backlin.cabgrid.res.in/sesameGWR/) has been developed This platform provides key insights into differentially expressed genes, transcription factors, miRNAs, and molecular markers like simple sequence repeats, single nucleotide polymorphisms, and insertions and deletions associated with both biotic and abiotic stresses.. The functional genomics information and annotations embedded in this web resource were predicted through RNA-seq data analysis. Considering the impact of climate change and the nutritional and medicinal importance of sesame, this study is of utmost importance in understanding stress responses. SesameGWR will serve as a valuable tool for developing climate-resilient sesame varieties, thereby enhancing the productivity of this ancient oilseed crop.
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  • 文章类型: Journal Article
    人心肌中每个细胞的排列被认为对于心脏组织的有效运动至关重要。我们研究了96孔微条图案板,以排列人诱导多能干细胞(iPSC)衍生的心肌细胞(CMs),类似于胎儿心肌.在微带图案板上培养的对齐CM(ACM)表现出病理学,运动功能,基因表达,与在平板上培养的未对齐的CM(FCM)相比,药物反应更接近成体细胞。我们使用这些ACM来评估药物的副作用和疗效,并确定这些是否与成人样反应相似。当肥厚型心肌病(HCM)患者的CM接种并培养在微条图案板上或在ACM顶部分层时,两组HCM显示心率增加和同步收缩,表明心脏功能改善。建议ACM可以作为成人样心肌细胞的代表细胞用于药物筛选,并以细胞片的形式移植用于心力衰竭的再生治疗。
    The alignment of each cell in human myocardium is considered critical for the efficient movement of cardiac tissue. We investigated 96-well microstripe-patterned plates to align human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs), which resemble fetal myocardium. The aligned CMs (ACMs) cultured on the microstripe-patterned plates exhibited pathology, motor function, gene expression, and drug response that more closely resembled those of adult cells than did unaligned CMs cultured on a flat plate (FCMs). We used these ACMs to evaluate drug side effects and efficacy, and to determine whether these were similar to adult-like responses. When CMs from patients with hypertrophic cardiomyopathy (HCMs) were seeded and cultured on the microstripe-patterned plates or layered on top of the ACMs, both sets of HCMs showed increased heart rate and synchronized contractions, indicating improved cardiac function. It is suggested that the ACMs could be used for drug screening as cells representative of adult-like CMs and be transplanted in the form of a cell sheet for regenerative treatment of heart failure.
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  • 文章类型: Journal Article
    简介:脓毒症是一种临床综合征,其特征是由于感染引起的宿主免疫反应失调,导致危及生命的器官损伤。尽管积极推广和实施早期预防措施和集束化治疗,脓毒症的发病率和死亡率仍然很高,没有最佳的药物干预.叶花素(LBT),在党参中发现的聚乙炔的关键成分,已被科学证明具有有效的抗氧化和抗肿瘤特性。然而,其对脓毒症的治疗潜力尚不清楚.方法:小鼠接受腹膜内注射LBT预处理,然后注射脂多糖(LPS)诱导脓毒症。收集外周血标本检测TNF-α,IL-1β,和IL-6水平。在不同的时间间隔记录不同组的生存状态。RNA-Seq用于分析用LBT或LPS处理的腹膜巨噬细胞中的基因表达。结果:在这项研究中,在LPS诱导的脓毒症小鼠模型中,我们观察到LBT预处理小鼠的存活率显着提高。LBT显示IL-6,TNF-α的产生显着减少,血清中的IL-1β,随着减轻肺和肝组织损伤的特点是减少的炎症细胞浸润。此外,通过RNA-seq分析以及GO和KEGG分析,研究表明,LBT有效抑制了与细菌存在相关的基因,细胞对脂多糖刺激的反应,以及涉及Cxcl10,Tgtp1,Gbp5,Tnf,Il1b和IRF7特异性地在巨噬细胞内。我们还证实,LBT显著下调IL-6、TNF-α的表达,和IL-1β在LPS诱导的巨噬细胞活化中的作用。讨论:因此,我们的发现表明,LBT有效抑制炎症细胞因子的产生(IL-6,TNF-α,和IL-1β),并通过调节巨噬细胞产生这些细胞因子的能力来减轻LPS诱导的脓毒症。这些结果表明LBT有望成为脓毒症治疗的潜在治疗剂。
    Introduction: Sepsis is a clinical syndrome characterized by dysregulation of the host immune response due to infection, resulting in life-threatening organ damage. Despite active promotion and implementation of early preventative measures and bundle treatments, sepsis continues to exhibit high morbidity and mortality rates with no optimal pharmacological intervention available. Lobetyolin (LBT), the crucial component of polyacetylenes found in Codonopsis pilosula, has been scientifically proven to possess potent antioxidant and antitumor properties. However, its therapeutic potential for sepsis remains unknown. Methods: The mice received pretreatment with intraperitoneal injections of LBT, followed by injection with lipopolysaccharide (LPS) to induce sepsis. Peripheral blood samples were collected to detect TNF-α, IL-1β, and IL-6 levels. The survival status of different groups was recorded at various time intervals. RNA-Seq was utilized for the analysis of gene expression in peritoneal macrophages treated with LBT or LPS. Results: In this study, we observed a significant increase in the survival rate of mice pretreated with LBT in LPS induced sepsis mouse model. LBT demonstrated a remarkable reduction in the production of IL-6, TNF-α, and IL-1β in the serum, along with mitigated lung and liver tissue damage characterized by reduced inflammatory cell infiltration. Additionally, through RNA-seq analysis coupled with GO and KEGG analysis, it was revealed that LBT effectively suppressed genes associated with bacterium presence, cellular response to lipopolysaccharide stimulation, as well as cytokine-cytokine receptor interaction involving Cxcl10, Tgtp1, Gbp5, Tnf, Il1b and IRF7 specifically within macrophages. We also confirmed that LBT significantly downregulates the expression of IL-6, TNF-α, and IL-1β in macrophage activation induced by LPS. Discussion: Therefore, our findings demonstrated that LBT effectively inhibits the production of inflammatory cytokines (IL-6, TNF-α, and IL-1β) and mitigates sepsis induced by LPS through modulating macrophages\' ability to generate these cytokines. These results suggest that LBT holds promise as a potential therapeutic agent for sepsis treatment.
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