Barley Protein LFBEP-C1 from Lactiplantibacillus plantarum dy-1 Fermented Barley Extracts by Inhibiting Lipid Accumulation in a Caenorhabditis elegans Model

Objective This study aimed to investigate the lipid-lowering activity of LFBEP-C1 in high glucose-fed Caenorhabditis elegans(C.elegans).Methods In this study,the fermented barley protein LFBEP-C1 was prepared and tested for its potential anti-obesity effects on C.elegans.The worms were fed Escherichia coli OP50(E.coli OP50),glucose,and different concentrations of LFBEP-C1.Body size,lifespan,movement,triglyceride content,and gene expression were analyzed.The results were analyzed using ANOVA and Tukey's multiple comparison test.Results Compared with the model group,the head-swing frequency of C.elegans in the group of LFBEP-C1 at 20μg/mL increased by 33.88%,and the body-bending frequency increased by 27.09%.This indicated that LFBEP-C1 improved the locomotive ability of C.elegans.The average lifespan of C.elegans reached 13.55 days,and the body length and width of the C.elegans decreased after LFBEP-C1 intake.Additionally,LFBEP-C1 reduced the content of lipid accumulation and triglyceride levels.The expression levels of sbp-1,daf-2,and mdt-15 significantly decreased,while those of daf-16,tph-1,mod-1,and ser-4 significantly increased after LFBEP-C1 intake.Changes in these genes explain the signaling pathways that regulate lipid metabolism.Conclusion LFBEP-C1 significantly reduced lipid deposition in C.elegans fed a high-glucose diet and alleviated the adverse effects of a high-glucose diet on the development,lifespan,and exercise behavior of C.elegans.In addition,LFBEP-C1 regulated lipid metabolism mainly by mediating the expression of genes in the sterol regulatory element-binding protein,insulin,and 5-hydroxytryptamine signaling pathways.

Pharmacokinetics of Gelsemium elegans in female rats

Background:Gelsemium elegans Benth(G.elegans)is a poisonous perennial evergreen vine plant that has been applied in livestock production and veterinary clinical practice.Early studies found that the toxicity of G.elegans showed significant gender differences in rats,but the underlying reasons for this difference are still not well understood.Methods:In order to explore whether the gender differences in the toxicity of G.elegans are related to pharmacokinetic differences,based on the previous pharmacokinetic study of multiple components of G.elegans in male rats,this study used HPLC-MS/MS method established in the laboratory to conduct a pharmacokinetic study of multiple alkaloids in the plasma of female rats after a single gavage administration of G.elegans(dose of 0.1 g/kg).Results:Through detection,17 alkaloid components in the plasma of female rats were identified,and the pharmacokinetic parameters of 11 of these alkaloids were calculated.We find that in female rats.The T_(max)values were generally less than 0.5 h,and the T_(1/2)values exceeded 3 h,with the longest reaching up to 32.80 h half elimination time.Additionally,the C_(max)and AUC results indicated that female rats had generally higher absorption and exposure levels for most alkaloids.Conclusion:These results suggest that the reason for the differences in the toxicology of G.elegans may be related to the absorption and exposure of gelsemidine-type alkaloids in animals.

南海海绵Dactylospongia elegans的二倍半萜类化学成分及抗炎活性研究

利用溶剂分步萃取、正相硅胶柱层析、反相ODS柱层析和高效液相等多种色谱技术对中国南海海绵Dactylospongia elegans的化学成分进行分离纯化,通过波谱学手段并结合文献比对鉴定化合物的结构。从二氯甲烷萃取部位分离鉴定了8个化合物:dactylospene F(1)、scalarin(2)、honulactone A(3)、honulactone B(4)、honulactone E(5)、honulactone F(6)、honulactone I(7)、honulactone J(8),其中化合物1为新化合物,化合物2~8为首次从该海绵中分离得到。对8个化合物的抗炎活性进行了评价,结果显示化合物1、3、4在10μmol/L时对脂多糖诱导的一氧化氮生成有较好的抑制作用,抑制率分别为65.5%、48.5%、46.0%,且对小鼠巨噬细胞RAW 264.7无细胞毒性。

Exploring the anti-aging effects of chlorogenic acid and the underlying mechanisms based on a Caenorhabditis elegans model

Objective:To explore the anti-aging effects of chlorogenic acid(CGA)and the underlying mechanisms based on a Caenorhabditis elegans(C.elegans)model.Methods:The anti-agingactivityofCGAwasstudied basedon thebodylength,exercisebehavior,lipofuscin content,antioxidative stress ability,swallowing frequency,body-bending frequency,and head-swinging ability of C.elegans.Through DAF-16 nuclear translocation and SOD-3-GFP fluorescence experiments,the effects of CGA on ROS levels,antioxidant enzyme activities,MDA content,mutant-strain lifespan,and anti-aging molecular signaling pathways were explored,as well as the underlying mechanisms.Results:CGA improved multiple indices of the nematode:body length was increased(all P<0.001),head-swing frequency and body-bending frequency were increased(all P<0.05),nematode longevity was prolonged(P=0.0021),lipofuscin deposition in nematodes was slowed down(all P<0.001),the chemotaxis index was improved(P=0.0012),ROS levels were reduced(all P<.001),and SOD activity and MDA content were reduced(SOD:P=0.0017 between the low-concentration group and the control group,P<.001 between the high-concentration and medium-concentration groups and the control group;MDA:P=0.0135 between the low-concentration group and the control group,and P<0.001 between the high-concentration and medium-concentration groups and the control group).In addition,CGA also activated the DAF-16 transcription factor,promoted DAF-16 nuclear translocation under oxidative stress conditions(both P<0.001 between the high-concentration and medium-concentration groups and the control group),and increased SOD-3 gene expression in nematodes(all P<0.001).Conclusion:CGA plays an anti-aging role in C.elegans.The underlying mechanisms include activation of the insulin/IGF-1 signaling pathway and enhancement of DAF-16 activity.This study lays a foundation for further research into the anti-aging effects of CGA.

The role of crm-1 in ionizing radiation-induced nervous system dysfunction in Caenorhabditis elegans

Ionizing radiation can cause changes in nervous system function.However,the underlying mechanism remains unclear.In this study,Coenorhabditis elegans(C.elegans)was irradiated with 75 Gy of ^(60)Co whole-body γ radiation.Behavioral indicators(head thrashes,touch avoidance,and foraging),and the development of dopaminergic neurons related to behavioral function,were evaluated to assess the effects of ionizing radiation on nervous system function in C.elegans.Various behaviors were impaired after whole-body irradiation and degeneration of dopamine neurons was observed.This suggests that 75 Gy of γ radiation is sufficient to induce nervous system dysfunction.The genes nhr-76 and crm-1,which are reported to be related to nervous system function in human and mouse,were screened by transcriptome sequencing and bioinformatics analysis after irradiation or sham irradiation.The expression levels of these two genes were increased after radiation.Next,RNAi technology was used to inhibit the expression of crm-1,a gene whose homologs are associated with motor neuron development in other species.Downregulation of crm-1 expression effectively alleviated the deleterious effects of ionizing radiation on head thrashes and touch avoidance.It was also found that the expression level of crm-1 was regulated by the nuclear receptor gene nhr-76.The results of this study suggest that knocking down the expression level of nhr-76 can reduce the expression level of crm-1,while down-regulating the expression level of crm-1 can alleviate behavioral disorders induced by ionizing radiation.Therefore,inhibition of crm-1 may be of interest as a potential therapeutic target for ionizing radiation-induced neurological dysfunction.

Metabolomics and gene expression levels reveal the positive effects of teaseed oil on lifespan and aging process in Caenorhabditis elegans

As an irreplaceable dietary constituent,lipids play a vital important role in health,but their effects on aging process and longevity are still not well known yet.In this paper,the metabolic profiling and gene expression levels of Caenorhabditis elegans were investigated to explore the effects of different edible oils on senescence and lifespan.The results showed that teaseed oil(TO)could prolong the life expectancy and slow down the aging process of C.elegans.Compared to the control group,the intake of lard oil(LO)and TO increased the expression levels of genes related to inhibition of protein aggregation(akt-1,daf-16,hsf-1,hsp-16.2)and lipid metabolism(daf-7,daf-1,mdt-15,lipl-4,fat-5,fat-6,fat-7),with a more significant alteration in TO group.Metabolomics revealed that palm oil can upregulated plenty of fatty acids(palmitic acid,stearic acid,tetracosanoic acid),together with some amino acids(tryptophan,L-aspartate,L-valine)and carbohydrate(D-glucose),while the trend was opposite in TO group.Besides,moderate-to-strong correlations were found between differential metabolites and changed genes.In general,this paper claimed that TO could prolong lifespan and slow down aging process via regulating the lipids,amino acids and carbohydrates metabolism.

Application of Caenorhabditis elegans in screening active ingredients of traditional Chinese medicine

People continue to recognize the important role of Chinese medicine in the treatment of diseases,and there is an urgent need to clarify the mechanism of Chinese medicine.Because of the multi-target and multi-component characteristics of traditional Chinese medicine,it is difficult to elaborate from a single perspective.As a high-quality animal model,Caenorhabditis elegans with the characteristics of small size and fast reproduction,has been widely used in traditional Chinese medicine activity experiments.In this article,Caenorhabditis elegans,as model organisms used to screen the active components of traditional Chinese medicine were reviewed.

Computational analysis of genetic loci required for amphid structure and functions and their possibly corresponding microRNAs in C. elegans

Objective To examine the important roles of microRNAs （miRNAs） in regulating amphid structure and function, we performed a computational analysis for the genetic loci required for the sensory perception and their possibly corresponding miRNAs in C. elegans. Methods Total 55 genetic loci required for the amphid structure and function were selected. Sequence alignment was combined with E value evaluation to investigate and identify the possible corresponding miRNAs. Results Total 30 genes among the 55 genetic loci selected have their possible corresponding regulatory miRNA（s）, and identified genes participate in the regulation of almost all aspects of amphid structure and function. In addition, our data suggest that both the amphid structure and the amphid functions might be regulated by a series of network signaling pathways. Moreover, the distribution of miRNAs along the 3＇ untranslated region （UTR） of these 30 genes exhibits different patterns. Conclusion We present the possible miRNA-mediated signaling pathways involved in the regulation of chemosensation and thermosensation by controlling the corresponding sensory neuron and interneuron functions. Our work will be useful for better understanding of the miRNA-mediated control of the chemotaxis and thermotaxis in C. elegans.

原位杂交检测mex-3 mRNA在C.elegans野生型胚胎发育中的分布

Caenorhabditiselegans是发育生物学中的重要模式生物,也是全基因序列已知的唯一多细胞动物.许多基因参与C.elegans的细胞命运决定和细胞谱系发育.mex 3是C.elegans早期胚胎发育中细胞命运决定的重要基因.本文报告采用原位杂交技术检测mex 3mRNA在C.elegans野生型胚胎发育中的分布,发现mex 3mRNA在生殖腺和卵母细胞的细胞质都有广泛而丰富的分布,从受精开始mex 3mRNA的分布开始发生了变化,从均一分布的模式开始进入不对称的分布模式,在2细胞阶段mex 3mRNA仅在前部的大卵裂球分布很多、染色较深,后部的小卵裂球低水平分布,染色较浅,在4细胞的早期胚胎中只在前部的两个卵裂球有较丰富的分布,后部的2个细胞低水平分布.到8细胞和16细胞阶段,只有后部的一个卵裂球检测到mRNA的分布,此后的胚胎中未检测到mRNA的分布.

细菌介导的RNA干扰对C.elegans中par-3基因的作用

设计并构建了针对par-3基因的发夹RNA载体,将构建好的质粒转入大肠杆菌HT115,25℃喂食Caenorhabditiselegans(C.elegans)野生型虫体,24h后观察par-3(RNA干扰)C.elegans的胚胎发育情况。结果显示通过喂食形成发夹结构dsRNA的细菌可以对C.elegans中par-3基因进行RNA干扰,干扰率可以达到60%以上。干扰后的早期胚胎发育丧失第一次卵裂的不对称性,第二次卵裂的纺锤体方向发生改变,与par-3突变体的观察结果一致,为进一步在mex-3转基因虫体中通过RNA干扰研究基因表达打下了基础。

Involvement of genes required for synaptic function in aging control in C.elegans

Objective To identify new genes required for neurosecretory control of aging in C. elegans. Methods In view of the importance of nervous system in aging regulation, we performed the screen for genes involved in the aging regulation from genetic loci encoding synaptic proteins by lifespan assay and accumulation of lipofuscin autofluorescence. We further investigated the dauer formation phenotypes of their corresponding mutants and whether they were possibly up-regulated by the insulin-like signaling pathway. Results The genetic loci of unc-10, syd-2, hlb-1, dlk-1, mkk-4, scd- 2, snb-1, ric-4, nrx-1, unc-13, sbt-1 and unc-64 might be involved in the aging control. In addition, functions of unc-10, syd-2, hlb-1, dlk-1, mkk-4, scd-2, snb-1, ric-4 and nrx-1 in regulating aging may be opposite to those of unc-13, sbt-1 and unc-64. The intestinal autofluorescence assay further indicated that the identified long-lived and short-lived mutants were actually due to the suppressed or accelerated aging. Among the identified genes, syd-2, hlb-1, mkk-4, scd-2, snb-1, ric-4 and unc-64 were also involved in the control of dauer formation. Moreover, daf-2 mutation positively regulated the expression of syd-2 and hlb-1, and negatively regulated the expression of mkk-4, nrx-1, ric-4, sbt-1, rpm-1, unc-10, dlk- 1 and unc-13. The daf-16 mutation positively regulated the expression of syd-2 and hlb-1, and negatively regulated the expression of mkk-4, nrx-1, sbt-1, rpm-1, unc-10, dlk-1 and unc-13. Conclusion These data suggest the possibly important status of the synaptic transmission to the animal＇ s life-span control machinery, as well as the dauer formation control.

原位杂交检测pal-1 mRNA在C.elegans野生型和突变体早期胚胎中的分布

Caenorhabditis elegans是发育生物学中的重要模式生物,许多基因参与C.elegans的细胞命运决定和细胞谱系发育.par基因的活性影响C.elegans胚胎前后轴的极性,其功能缺失会使胚胎的第一次分裂丧失前后轴的不对称性,导致一些母体提供的发育调控因子不能在特定的胚胎细胞中准确定位,从而改变胚胎细胞发育命运.pal-1是C.elegans早期胚胎发育中决定细胞命运的重要基因,它决定体细胞的性质,也是转录因子,调控着后续基因的表达,凡含有该基因表达的细胞发育成体细胞.调控PAL-1表达的基因有许多,本文通过原位杂交检测pal-1 mRNA在C.elegans野生型和par-5、par-6、gld-1突变体早期胚胎中的分布,来探讨par-5、par-6和gld-1基因的活性在胚胎发育早期对pal-1 mRNA的影响.实验结果表明,pal-1mRNA在野生型早期胚胎中有明显极性,而pal-1mRNA分布的不对称性在上述基因缺失的情况下被破坏,这些基因与pal-1mRNA的不对称分布直接相关.

原位杂交检测lin-28 mRNA在Caenorhabditis elegans中的分布

利用原位杂交检测了异时性基因lin-28 mRNA在野生型和lin-4,lin-14突变体以及lin-4,lin-14双突变体中的分布,发现lin-28 mRNA在胚胎和成虫生殖腺中存在.还发现lin-4突变不改变lin-28 mRNA分布,证明了lin-28表达独立于lin-4抑制通路的存在.

Screening for genetic loci affecting the active zone formation in C. elegans

Objective To screen and identify genetic loci affecting the active zone formation in C. elegans. Methods A SYD-2：：GFP reporter was constructed and used as an active zone marker for forward genetic screen to identify genetic loci affecting the active zone formation. Results Eight isolated mutant alleles were characterized from 15,000 haploid genomes. The SYD-2：：GFP phenotypes of these mutants are mainly reflected as the changes of number, morphology, distribution of puncta and the gaps appearance. Some mutants also exhibit visible behavioral or physical phenotypes, and aldicarb resistant or sensitive phenotypes. Conclusion These mutants provide the opportunity for further systematic research on the active zone formation and the neurotransmission.

食细菌线虫Caenorhabditis elegans的取食偏好性

通过设置平板培养试验,以模式种线虫Caenorhabditis elegans为材料,观察了食细菌线虫的取食行为。结果表明:C.elegans在取食细菌时对原位土壤中分离的一种Pseudomonas sp细菌存在最大的取食偏好性。这种取食偏好性表现在大部分C.elegans在24 h内都直接朝Pseudomonas sp迁移,说明C.elegans能通过某种机制识别Pseudomonas sp。距离迁移试验及C.elegans迁移率表明它可能是通过辨别细菌所发出的气味识别它喜欢的食物。C.elegans的繁殖率跟其取食的偏好性是相关的,在迁移率较高的细菌培养基中线虫表现出更高的繁殖率。结果还表明:相对于G+细菌而言,C.elegans偏好取食G-细菌。为进一步了解土壤生态系统中食细菌线虫与细菌群落结构间相互关系提供了帮助。

ATP depletion is the major cause of MPP^+ induced dopamine neuronal death and worm lethality inα-synuclein transgenic C.elegans

Objective To investigate the toxic effect of environmental neurotoxin MPP^＋ to C.elegans and identify the mechanisms that cause the toxicity.Methods Humanα-synuclein transgenic C.elegans was used as the animal model,the toxic effect of MPP^＋ to dopamine（DA）neurons and the lifespan of worms was tested.The worms were feed with OP50 to determine whether ATP increase can rescue the worm from toxicity.ATP level and aberrant protein accumulation were analyzed in the MPP^＋ treated worms with or without OP50 addition.Results We found that MPP^＋ induced DA cell death and worm lethality,which could be prevented by OP50 treatment.OP50 exerted the protective effect by up-regulating ATP level,even though it also induced accumulation ofα-synuclein.Despite the undefined role of protein aggregation to the cell death,our results showed that the toxicity of MPP^＋ was mainly caused by the ATP depletion in theα-synuclein transgenic C.elegans.Conclusion MPP^＋ could induce DA neuronal death and worm lethality inα-synuclein transgenic C.elegans;Compared with the aggregation ofα-synuclein,the major cause of MPP^＋ toxicity appeared due to ATP depletion.

Toxicity of Selected Imidazolium-based Ionic Liquids on Caenorhabditis elegans： a Quantitative Structure-Activity Relationship Study

Due to the large number of ionic liquids （ILs） and their potential environmental risk, assessing the toxicity of ILs by ecotoxicological experiment only is insufficient. Quantitative structure- activity relationship （QSAR） has been proven to be a quick and effective method to estimate the viscosity, melting points, and even toxicity of ILs. In this work, the LC50 values of 30 imidazolium-based ILs were determined with Caenorhabditis elegans as a model animal. Four suitable molecular descriptors were selected on the basis of genetic function approximation algorithm to construct a QSAR model with an R^2 value of 0.938. The predicted lgLC50 in this work are in agreement with the experimental values, indicating that the model has good stability and predictive ability. Our study provides a valuable model to predict the potential toxicity of ILs with different sub-structures to the environment and human health.

Learning and learning choice in the nematode Caenorhabditis elegans

The nematode Caenorhabditis elegans is an attractive model organism to study the behavioral plasticity for its simple system and ability to respond to diverse environmental stimuli, such as touch, smell, taste and temperature. Learning in C. elegans encompasses both non-associative learning and associative learning. Till now, themotaxis and chemotaxis are two major paradigms for associative learning and there are at least 6 forms of chemotaxis-mediated associative learning. Three research systems have also been explored to study the mechanism of learning choice in worms. This review will discuss the forms, research models, genetic and molecular regulation of learning and learning choice in C. elegans.

C.elegans野生型和par突变体早期胚胎卵裂的观察

不对称分裂在动植物的发育中起到了非常重要的作用。Caenorhabditiselegans(C.elegans)胚胎最早的两次卵裂为研究控制不对称分裂的机制提供了很好的机会。用普通光学显微镜观察了野生型胚胎早期卵裂和par-1、par-2、par-3、par-4突变体胚胎的早期卵裂。野生型胚胎最早的分裂是不等的,产生了两个不同大小的子细胞。两个子细胞又以不同的方向进行第二次分裂。在C.elegans中任意一个par基因的缺失会使胚胎的第一次卵裂丧失不对称性。这会导致一些发育调控因子不能在特定的胚胎细胞中准确地定位,造成细胞分裂纺锤体方向的异常。par类基因参与不对称性的建立,这种不对称性决定了C.elegans身体的前后轴。

原位杂交检测lin-4 mRNA在Caenorhabditis elegans中的表达

lin-4是控制Caenorhabditis elegans(C.elegans)幼虫发育的异时性基因,也是一种小RNA分子(microRNA)。通过整体原位杂交检测小RNA lin-4在野生型和lin-14、lin-28突变体中的区域性表达,探讨lin-4在C.elegans发育时空控制中的作用。结果表明:lin-4 mRNA在胚胎发育的早期和中期表达,胚胎后期至L1期末没有表达,之后又持续表达,成虫中也可以检测到lin-4 mRNA的存在。在lin-14和lin-28突变体中,lin-4的表达基本与野生型一致,不受lin-14、lin-28基因突变的影响,说明lin-14和lin-28是lin-4的下游基因。