Novel SNP of EPAS1 gene associated with higher hemoglobin concentration revealed the hypoxia adaptation of yak(Bos grunniens)

Endothelial PAS domain protein 1 gene （EPAS1） is a key transcription factor that activates the expression of oxygen-regu- lated genes. In this study, in order to better understand the effects of EPAS1 gene on hematologic parameters in yak, we firstly quantified the tissue expression patterns for EPASl mRNA of yak, identified polymorphism in this gene and evaluated its association with hematologic parameters. Expression of EPAS1 mRNA was detected in all eight tissues （heart, liver, lung, spleen, pancreas, kidney, muscles and ovary）. The expressions of EPAS1 in lung and pancreas were extremely higher than other tissues examined. Three novel single nucleotide polymorphisms （SNPs） （g.83052 C〉T, g.83065 G〉A and g.83067 C〉A） within the EPAS1 were identified and genotyped in Pali （PL）, Gannan （GN） and Tianzhu White （TZW） yak breeds. Significant higher frequencies of the AA and GA genotypes and A allele of the g.83065 G〉A were observed in the PL and GN breeds than that in the TZW breed （P〈0.01）. Association analysis of the PL breed indicated that the g.83065 G〉A polymorphism was significantly associated with hemoglobin （HGB） concentration in yaks （P〈0.05）. Individuals with genotype AA had significantly higher HGB concentration （P〈0.05） than those with genotype GA and GG. All these results will help our further understanding of biological functional of yak EPAS1 gene in responding to hypoxia and also indicate EPAS1 might contribute to the hypoxia adaptation of the yak.

Gene expression of vascular endothelial growth factor A and hypoxic adaptation in Tibetan pig

Background： Vascular endothelial growth factor A （VEGFA） can induce endothelial cell proliferation, promote cell migration, and inhibit apoptosis. These processes play key roles in physiological blood vessel formation and pathological angiogenesis. Methods： In this study, we examined VEGFA gene expression in the heart, liver, and kidney of Tibetan pigs （-I-P）, Yorkshire pigs that migrated to high altitudes （YH）, and Yorkshire pigs that lived at low altitudes （YL）. We used PCR and Sanger sequencing to screen for single nucleotide polymorphisms （SNPs） in 5＇-flanking DNA and exons of the VEGFA gene. Quantitative real-time PCR and western blots were used to measure expression levels and PCR products were sequenced. Results： Results showed that the VEGFA mRNA and protein expression in heart, liver and kidney of TP was higher than that in YH and YL. In addition, the mRNA sequence of the pig VEGFA gene was conserved among pig breeds, and only five SNPs were found in the 5＇-flanking region of the VEGFA gene, the allele frequency distributions of the 5 SNPs were not significantly different between the TP, Yorkshire （YL）, and Diannan small-ear （DN） pig populations. Conclusion： In conclusion, the Tibetan pig showed high tissues, which suggests that the VEGFA gene may play a levels of VEGFA gene expression in several hypoxic major functional role in hypoxic adaptation.

Genome-wide DNA methylation landscape of four Chinese populations and epigenetic variation linked to Tibetan high-altitude adaptation

DNA methylation(DNAm)is one of the major epigenetic mechanisms in humans and is important in diverse cellular processes.The variation of DNAm in the human population is related to both genetic and environmental factors.However,the DNAm profiles have not been investigated in the Chinese population of diverse ethnicities.Here,we performed double-strand bisulfite sequencing(DSBS)for 32 Chinese individuals representing four major ethnic groups including Han Chinese,Tibetan,Zhuang,and Mongolian.We identified a total of 604,649 SNPs and quantified DNAm at more than 14 million Cp Gs in the population.We found global DNAm-based epigenetic structure is different from the genetic structure of the population,and ethnic difference only partially explains the variation of DNAm.Surprisingly,non-ethnic-specific DNAm variations showed stronger correlation with the global genetic divergence than these ethnic-specific DNAm.Differentially methylated regions(DMRs)among these ethnic groups were found around genes in diverse biological processes.Especially,these DMR-genes between Tibetan and nonTibetans were enriched around high-altitude genes including EPAS1 and EGLN1,suggesting DNAm alteration plays an important role in high-altitude adaptation.Our results provide the first batch of epigenetic maps for Chinese populations and the first evidence of the association of epigenetic changes with Tibetans'high-altitude adaptation.

Genome and population evolution and environmental adaptation of Glyptosternon maculatum on the Qinghai-Tibet Plateau

Persistent uplift means the Qinghai-Tibet Plateau(QTP)is an ideal natural laboratory to investigate genome evolution and adaptation within highland environments.However,how paleogeographic and paleoclimatic events influence the genome and population of endemic fish species remains unclear.Glyptosternon maculatum is an ancient endemic fish found on the QTP and the only critically endangered species in the Sisoridae family.Here,we found that major transposons in the G.maculatum genome showed episodic bursts,consistent with contemporaneous geological and climatic events during the QTP formation.Notably,histone genes showed significant expansion in the G.maculatum genome,which may be mediated by long interspersed nuclear elements(LINE)repetitive element duplications.Population analysis showed that ancestral G.maculatum populations experienced two significant depressions 2.6 million years ago(Mya)and 10000 years ago,exhibiting excellent synchronization with Quaternary glaciation and the Younger Dryas,respectively.Thus,we propose that paleogeography and paleoclimate were dominating driving forces for population dynamics in endemic fish on the QTP.Tectonic movements and temperature fluctuation likely destroyed the habitat and disrupted the drainage connectivity among populations.These factors may have caused severe bottlenecks and limited migration among ancestral G.maculatum populations,resulting in the low genetic diversity and endangered status of the species today.

HAL1 mediate salt adaptation in Arabidopsis thaliana

The yeast HAL1 gene was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens-mediated transformation with vacuum infiltration under the control of CaMV 35S promoter. Thirty-three individual kanamycin resistant plants were obtained from 75,000 seeds. Southern blotting analysis indicated that HAL1 gene had been integrated into all of the transgenic plants’ genomes. The copy number of HAL1 gene in transgenic plants was mostly 1 to 3 by Southern analysis. Phenotypes of transgenic plants have no differences with wild type plants. several samples of transformants were self-pollinated, and progenies from transformed and non-transformed plants (controls) were evaluated for salt tolerance and gene expression. Measurement of concentrations of intracellular K+ and Na+ showed that transgenic lines were able to retain less Na+ than that of the control under salt stress. Results from different tests indicated the expression of HAL1 gene promotes a higher level of salt tolerance in vivo in the transgenic Arabidopsis plants.

Perspectives on studying molecular adaptations of amphibians in the genomic era

Understanding the genetic mechanisms underlying particular adaptations/phenotypes of organisms is one of the core issues of evolutionary biology.The use of genomic data has greatly advanced our understandings on this issue,as well as other aspects of evolutionary biology,including molecular adaptation,speciation,and even conservation of endangered species.Despite the well-recognized advantages,usages of genomic data are still limited to non-mammal vertebrate groups,partly due to the difficulties in assembling large or highly heterozygous genomes.Although this is particularly the case for amphibians,nonetheless,several comparative and population genomic analyses have shed lights into the speciation and adaptation processes of amphibians in a complex landscape,giving a promising hope for a wider application of genomics in the previously believed challenging groups of organisms.At the same time,these pioneer studies also allow us to realize numerous challenges in studying the molecular adaptations and/or phenotypic evolutionary mechanisms of amphibians.In this review,we first summarize the recent progresses in the study of adaptive evolution of amphibians based on genomic data,and then we give perspectives regarding how to effectively identify key pathways underlying the evolution of complex traits in the genomic era,as well as directions for future research.

Gene cloning and sequence analysis of the cold-adapted chaperones DnaK and DnaJ from deep-sea psychrotrophic bacterium Pseudoalteromonas sp. SM9913

Pseudoalteromonas sp. SM9913 is a phychrotmphic bacterium isolated from the deep-sea sediment. The genes encoding chaperones DnaJ and DnaK of P. sp. SM9913 were cloned by normal PCR and TAIL - PCR （GenBank accession Nos DQ640312, DQ504163 ）. The chaperones DnaJ and DnaK from the strain SM9913 contain such conserved domains as those of many other bacteria, and show some cold-adapted characteristics in their structures when compared with those from psychro-, meso-and themophilic bacteria. It is indicated that chaperones DnaJ and DnaK of P. sp. SM9913 may be adapted to low temperature in deep-sea and function well in assisting folding, assembling and translocation of proteins at low temperature. This research lays a foundation for the further study on the cold-adapted mechanism of chaperones DnaJ and DnaK of cold-adapted microorganisms.

Complete genome analysis of bacteriochlorophyll acontaining Roseicitreum antarcticum ZS2-28^(T)reveals its adaptation to Antarctic intertidal environment

Aerobic anoxygenic phototrophic bacteria(AAPB)are photoheterotrophic prokaryotes able to use both light and dissolved organic matter as energy sources.Roseicitreum antarcticum ZS2-28^(T)was isolated from intertidal sediment in the Larsemann Hills,Princess Elizabeth Land,Antarctica,and was able to produce bacteriochlorophyll a.It is the type strain of the sole species within the genus Roseicitreum.The complete genome sequence of the bacterium was determined using Illumina HiSeq X and PacBio RSII systems.The genome of R.antarcticum ZS2-28^(T)was 4253095 bp and consisted of one chromosome and four plasmids.A number of genes related to the bacteriochlorophyll a production,photosynthetic reaction,cold adaptation,salt adaptation,ultra-violet resistance and DNA damage repairing were found in the genome.In addition to genomic islands and typeⅣsecretion systems,genes related to gene transfer agents were detected in the genome of R.antarcticum ZS2-28^(T),suggesting that this bacterium can adapt to its environment by acquiring exogenous nucleic acids.The annotated complete genome sequence provides genetic insights into the environmental adaptation and ecological function of R.antarcticum ZS2-28^(T)in Antarctic coastal area.

A de novo assembled genome of the Tibetan Partridge (Perdix hodgsoniae) and its high-altitude adaptation

The Tibetan Partridge(Perdix hodgsoniae)is an endemic species distributed in high-altitude areas of 3600–5600 m on the Qinghai–Tibet Plateau.To explore how the species is adapted to the high elevation environment,we as-sembled a draft genome based on both the Illumina and PacBio sequencing platforms with its population genetics and genomics analysis.In total,134.74 Gb short reads and 30.81 Gb long reads raw data were generated.The 1.05-Gb assembled genome had a contig N50 of 4.56 Mb,with 91.94%complete BUSCOs.The 17457 genes were annotated,and 11.35%of the genome was composed of repeat sequences.The phylogenetic tree showed that P.hodgsoniae was located at the basal position of the clade,including Golden Pheasant(Chrysolophus pictus),Common Pheasant(Phasianus colchicus),and Mikado Pheasant(Syrmaticus mikado).We found that 1014,2595,and 2732 of the 6641 one-to-one orthologous genes were under positive selection in P.hodgsoniae,detected using PAML,BUSTED,and aBSREL programs,respectively,of which 965 genes were common under positive selec-tion with 3 different programs.Several positively selected genes and immunity pathways relevant to high-altitude adaptation were detected.Gene family evolution showed that 99 gene families experienced significant expansion events,while 6 gene families were under contraction.The total number of olfactory receptor genes was relatively low in P.hodgsoniae.Genomic data provide an important resource for a further study on the evolutionary history of P.hodgsoniae,which provides a new insight into its high-altitude adaptation mechanisms.

Genetics of carotenoids for provitamin A biofortification in tropical-adapted maize

Yellow maize contains high levels of β-carotene(βC), making it an important crop for combating vitamin A deficiency through biofortification. In this study, nine maize inbred lines were selected at random from 31 provitamin A(PVA) maize inbred lines and crossed in a partial diallel mating design to develop 36 crosses. The crosses were evaluated in the field in two locations(Samaru and Kerawa) and their seed carotenoid content were determined by high-performance liquid chromatography. The modes of gene action, heritability, and correlations between agronomic traits and carotenoid content were estimated. Additive genetic variances(σ~2a) were lower than non-additive genetic variances(σ~2d) for all the carotenoids, plant height(PH), and grain yield(GY), suggesting a preponderance of non-additive gene action. Broad-sense heritability(H^2) was high(H^2> 60%) for zeaxanthin,days to anthesis, and PH, moderate(30% < H^2< 60%) for lutein and GY, and low(H^2< 30%)for alpha carotene, beta cryptoxanthin, βC, and PVA. Genetic advance as a percentage of mean, considered with H^2, also suggests a preponderance of non-additive gene action for PVA carotenoids. Hybrid variety development is thus an appropriate approach to improving grain yield and PVA. GY showed no significant genotypic correlations with carotenoid content, suggesting that these traits can be improved concurrently. Thus, there is ample scope for improvement of PVA and GY in the sample of tropical-adapted maize.

Comparative DNA methylation reveals epigenetic adaptation to high altitude in snub-nosed monkeys

DNA methylation plays a crucial role in environmental adaptations.Here,using whole-genome bisulfite sequencing,we generated comprehensive genome-wide DNA methylation profiles for the high-altitude Yunnan snub-nosed monkey(Rhinopithecus bieti)and the closely related golden snub-nosed monkey(R.roxellana).Our findings indicated a slight increase in overall DNA methylation levels in golden snub-nosed monkeys compared to Yunnan snub-nosed monkeys,suggesting a higher prevalence of hypermethylated genomic regions in the former.Comparative genomic methylation analysis demonstrated that genes associated with differentially methylated regions were involved in membrane fusion,vesicular formation and trafficking,hemoglobin function,cell cycle regulation,and neuronal differentiation.These results suggest that the high-altitude-related epigenetic modifications are extensive,involving a complete adaptation process from the inhibition of single Ca^(2+)channel proteins to multiple proteins collaboratively enhancing vesicular function or inhibiting cell differentiation and proliferation.Functional assays demonstrated that overexpression or down-regulation of candidate genes,such as SNX10,TIMELESS,and CACYBP,influenced cell viability under stress conditions.Overall,this research suggests that comparing DNA methylation across closely related species can identify novel candidate genomic regions and genes associated with local adaptations,thereby deepening our understanding of the mechanisms underlying environmental adaptations.

基于AGEP-DNN的水下聚能装药比冲量预测模型

聚能装药比冲量是表征水下爆炸中冲击波对目标破坏作用的重要参数。为实现水下聚能装药比冲量智能预测,提出一种自适应基因表达式编程(adaptive gene expression programming, AGEP)优化深度神经网络(deep neural network, DNN)的聚能装药比冲量预测模型(AGEP-DNN)。考虑装药结构与比冲量数值之间的复杂非线性关系,通过AUTODYN软件建立有限元模型,对水下爆炸过程进行仿真,采用经验公式验证仿真数据的有效性;基于仿真实验数据,设计AGEP算法优化DNN超参数,构建AGEP-DNN模型,对比冲量进行智能预测。实验结果显示,AGEP-DNN聚能装药比冲量预测模型在9种对比智能预测模型中具有最优的预测精度。

湘赣边区红色文化创意产品的适应性开发与设计

设计赋能文化传承的背景下,文化创意产品地域文化转译的同质化、传播力的疲软化制约了文化传承。本研究以湘赣边区红色文化创意产品设计为例,提出文化创意产品适应性开发方法,即凸显湘赣边区的红色文化,精准文化转译;匹配红色旅游群体的差异化需求,提升文化传播力。首先,以大学生群体为研究对象,依据用户消费类别偏好量表和产品需求特征量表探测用户需求。然后,解析湘赣边区红色文化基因,指导产品设计实践。最后,从设计层、功能层、文化层对设计产品展开后期评估,实证文化创意产品适应性开发方法的可行性。研究结果表明,湘赣边区红色文化创意产品的适应性开发方法能够辅助设计师的设计决策。

花斑裸鲤SIRT基因家族分析及低温适应性表达

花斑裸鲤是裂腹鱼亚科的代表物种之一,能够很好地适应青藏高原寒冷的环境。为探究sirtuins(SIRT)基因家族在花斑裸鲤低温适应中的作用,笔者对SIRT基因家族的7个基因进行了生物信息学分析和基因表达研究。结果显示,花斑裸鲤SIRT基因家族的长度差异较大,序列最长的是SIRT1基因(22885 bp),最短的是SIRT4基因(3660 bp),包含蛋白质编码区,编码304至691个氨基酸不等。通过保守结构域和基序分析发现,花斑裸鲤SIRT基因均具有Sir2保守结构域和基序,包括GAGxSx、xxPxxR、PxxxH和QNxDxLx。氨基酸理化性质预测结果表明,花斑裸鲤SIRT蛋白均为亲水性蛋白,除SIRT4之外,均是不稳定蛋白。亚细胞定位预测结果显示,SIRT蛋白主要分布于细胞质和细胞核中。通过预测蛋白质相互作用发现,SIRTs与SOD2、CAT、PPARGC1α、p53、FOXO1a和FOXO1b具有相互作用。进一步对SIRT基因在低温适应的花斑裸鲤的肝脏、脑和肌肉组织中表达进行分析,结果显示,低温下,SIRT2、SIRT4和SIRT7基因在肝脏和脑中表达显著上调,SIRT2~4、SIRT6和SIRT7基因在肌肉中表达显著上调,推测SIRT基因家族尤其是SIRT2、SIRT4和SIRT7基因在花斑裸鲤低温适应中可能具有重要作用。本试验分析了花斑裸鲤SIRT基因家族的特征,其结果可为后续SIRT基因在花斑裸鲤低温适应中功能研究提供基础。

野生大豆功能基因的研究进展

野生大豆(Glycine soja Sieb.&Zucc.)是栽培大豆(Glycine max [L.] Merr.)的近缘祖先种。在大豆驯化的过程中,栽培大豆丢失了大量的基因或等位变异,导致栽培大豆的遗传多样性降低,这严重限制了栽培大豆品种选育和改良的有效性与丰富性。我国野生大豆种质资源丰富,蕴藏着许多高蛋白含量、抗病虫、耐干旱、耐盐碱等方面的潜力基因,挖掘潜力基因并利用分子设计育种技术应用到现代的栽培大豆品种中,能够有效地拓宽栽培大豆的遗传多样性。本文综述了野生大豆的分布规律和形态特征、近年来在野生大豆中发掘的重要功能基因或位点,包括百粒重、开花期和成熟期、蛋白质和油分含量、抗病、抗虫、耐盐碱、耐干旱等重要农艺性状基因,并讨论这些重要基因或位点在未来栽培大豆育种中的应用潜力,以期为育种家培育和改良大豆新品种提供一种新的育种思路和策略。

New dating indicates intermittent human occupation of the Nwya Devu Paleolithic site on the high-altitude central Tibetan Plateau during the past 45,000 years

The timing and mechanisms of the human occupation of the demanding high-altitude Tibetan Plateau environment are of great interest.Here,we report on our reinvestigations and dating of the Nwya Devu site,located nearly 4600 meters above sea level on the central Tibetan Plateau.A new microblade techno-complex was identified on a lower lake shore at this site,distinct from the previously reported blade tool assemblage.These two lithic assemblages were dated to 45.6±2.6 and10.3±0.5 ka using optically stimulated luminescence and accelerator mass spectrometry^(14)C methods.They represent,respectively,the earliest known Paleolithic and microlithic sites on the interior Tibetan Plateau,indicating multiple occupation episodes of hunter-gatherers during the past 45 ka.Our studies reveal that relatively stable depositional conditions and a paleoenvironment characterized by a comparatively warm climate facilitated these multiple occupations at Nwya Devu.The contemporaneous occurrence of the Upper Paleolithic blade technology on the Tibetan Plateau and most of Eurasia between 50 and 40 ka indicates rapid,large-scale dispersals of humans that profoundly affected human demography on a large scale.Combining new archaeological evidence and previously reported genetic data,we conclude that the Tibetan Plateau provided a relatively stable habitat for Upper Paleolithic hunter-gatherers,which may have contributed to the complex and multiple-origin gene pool of present-day Tibetans.

基于小型化Cas蛋白的CRISPR/Gal4BD-Cas供体适配基因编辑系统研究

在哺乳动物细胞中,利用同源引导修复(homology-directed repair,HDR)机制的基因编辑策略能够实现精准的点编辑和敲入,但是HDR的低效性严重制约了该策略在精准医疗和分子设计育种中的应用。鉴于HDR机制所需的供体DNA模板不能自主募集到基因组双链断裂(double-stranded break,DSB)处,本课题组提出了供体适配系统(donor adapting system,DAS)的概念并开发出CRISPR/SpCas9-Gal4BD供体适配基因编辑系统。由于SpCas9蛋白分子较大,与Gal4BD适配器结合不利于表达、病毒载体包装及活体递送等过程,因此本研究进一步采用两种小型化Cas蛋白——路邓葡萄球菌(Staphylococcus lugdunensis)源SlugCas9变体SlugCas9-HF与氨基酸球菌(Acidaminococcus sp.)源AsCas12a,开发了新型的CRISPR/Gal4BD-SlugCas9和CRISPR/Gal4BD-AsCas12a供体适配基因编辑系统。通过SSA活性报告实验初步证明了Gal4BD与SlugCas9、AsCas12a N-端融合对其打靶活性影响较小。通过HDR效率报告实验进行功能验证并优化供体设计方案,结果表明:对于CRISPR/Gal4BD-AsCas12a DAS,适配器结合序列(binding sequence,BS)与供体5′-端融合(BS-dsDNA)效果较好;对于CRISPR/Gal4BD-SlugCas9 DAS,则BS与供体3′-端融合(dsDNA-BS)较佳。最终,利用CRISPR/Gal4BD-SlugCas9 DAS对HEK293T细胞中的EMX1、NUDT5、AAVS1三个基因位点分别实现了24%、37%、31%的精准编辑,相比对照组得到了显著提高。本研究为供体适配基因编辑系统的进一步优化提供了参考和借鉴,为后续动物分子设计育种应用研究提供了新的基因编辑工具。

不同CRISPR/Cas9供体适配基因编辑系统的比较及优化研究

在哺乳动物细胞中进行基因敲入通常采用同源定向修复(homology-directed repair,HDR)机制将外源DNA模板整合到目标基因组靶点中。然而HDR效率往往较低,其中外源DNA模板与目标基因组靶点的共定位是关键限制因素之一。为提高CRISPR/Cas9系统介导的HDR效率,本团队及前人研究将不同接头蛋白与SpCas9蛋白融合表达,利用其与特异性DNA序列结合的特性,构建了多种CRISPR/SpCas9供体适配基因编辑系统。为了便于比较、优化不同CRISPR/Cas9供体适配系统,本研究利用这些系统在HEK293T细胞GAPDH和ACTB基因末位外显子3′-端进行了eGFP基因敲入,并采用了优化的供体DNA模板设计方式,通过PCR和Sanger测序检测敲入的准确性,流式细胞分析进行敲入效率的检测。结果表明,将yGal4BD、hGal4BD、hLacI、hTHAP11和N57等接头蛋白与SpCas9蛋白C-端融合对其活性均无显著性影响;在GAPDH位点上,SpCas9融合yGal4BD、hGal4BD、hLacI和hTHAP11的供体适配系统等均能显著提高敲入效率;在ACTB位点上,SpCas9融合yGal4BD和hGal4BD能显著提高敲入效率;且增加供体DNA模板中的结合序列(binding sequence,BS)数量,有利于提高SpCas9-hTHAP11系统介导的敲入效率。总之,本研究比较并优化了不同的CRISPR/Cas9供体适配基因编辑系统,为后续相关的基因编辑应用研究提供了参考和借鉴。

纺织典型装备故障多特征自适应提取方法

化纤高速卷绕头故障特征提取是卷绕头故障诊断中的关键步骤。为解决化纤卷绕头故障诊断精度不高、可解释性差的难点,提出了数据驱动的化纤卷绕头故障多特征自适应提取方法。通过改进型基因表达式编程(GEP)的故障特征生成方法,设计了一种运算符与变量符随机组合编码、对位匹配与倒序运算解码方法,构建了突变、插串、重组的遗传算子,实现了多个故障特征构建与生成;提出了低冗余、高互补的多特征提取与分析方法,实现了特征间关系的可解释性分析与关键特征优选。实验结果表明:采用所提出的改进型GEP方法与传统特征提取方法、通用GEP方法所生成的故障特征进行对比,在线速度为1000、2000和3000 m/min状态下,卷绕头故障诊断精度分别提升了8.959%、3.87%、3.77%和2.601%、3.2%、2.018%,有效解决了卷绕头故障特征提取的难题;进一步的特征工程实验表明,所提方法对于多特征组合下的卷绕头故障关键特征提取具有较强的适应性能。

中脑星形胶质细胞神经营养因子在利福平诱导的小鼠胆汁淤积性肝损伤中的作用

目的探究中脑星形胶质细胞神经营养因子(mesencephalic astrocyte-derived neurotrophic factor,MANF)在利福平(rifampicin,RFP)诱导的胆汁淤积性肝损伤中的作用。方法借助肝细胞MANF特异性敲除(hepatocyte-specific MANF knockout,HKO)小鼠模型,观察MANF基因缺失对RFP诱导的小鼠胆汁酸转运体表达的影响。体外观察MANF敲减后对人肝癌细胞HepG2的核因子红细胞系2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)的影响。结果与野生型(wild type,WT)小鼠相比,RFP处理的WT小鼠的胆盐输出泵(bile salt export pump,BSEP)以及多药耐药相关蛋白4(multidrug resistance-associated protein 4,MRP4)的蛋白和基因表达水平增加。然而,与RFP处理的WT小鼠相比,HKO小鼠中BSEP、多药耐药蛋白1(multidrug resistance protein 1,MDR1)、MRP2/3/4和有机溶质转运蛋白α(organic solute transporterα,OSTα)的蛋白和(或)基因表达水平明显降低。体外细胞MANF敲减会削弱RFP诱导的Nrf2的表达以及核内移。结论MANF可能通过调节Nrf2的表达调控BAT的适应性表达,在RFP诱导的肝损伤中发挥保护作用。