Development of a Zebrafish Model for Rapid Drug Screening against Alzheimer＇s Disease

Alzheimer＇s disease, the leading cause of dementia in the elderly, is a complex neurodegenerative disorder which leads to a progressive decline in cognitive functions. A rapid screening model is highly demanded for identification and evaluation of novel anti-Alzheimer＇s disease drugs from a large numbers of compounds. Until now, numerous studies utilized zebrafish model for drug discovery. Since aluminum can induce a similar biological activity in zebrafish as in Alzheimer patients, in this study, we developed a novel animal model using 3 to 5 day post-fertilization larval zebrafish by optimizing the doses and duration of aluminum chloride exposure. Six anti-Alzheimer＇s disease drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model. Importantly, Rivastigmine, ThT, Flurbiprofen and AM-117 could increase the value of Dyskinesia Recovery Rate by 53.4-64%, 169.4-200%, 54.5-96% and 70.9-121%, respectively. Rivastigmine, Memantine, ThT, Flurbiprofen, Rosiglitazone and AM-117 improved the value of Response Efficiency by 86.6-175.1%, 28.2-66.6%, 127.2-236.5%, 118.3-323.7%, 26.6-140.8% and 70.2-161.4%, respectively. Our results suggest that the zebrafish model developed in this study could be a useful tool for high throughput screening of potential novel anti-Alzheimer＇s disease leading compounds targeting acetylcholinesterase, N-methyl-D-aspartic acid receptor, γ-secretase, peroxisome proliferator-activated receptor-γand amyloid-β.

Analysis of the key networks,metabolic pathways,and regulation substances of hypoxia based on the omics and zebrafish model

OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular mechanism of hypoxia still remains unclear.In this work,we further explore the molecular mechanism of hypoxia and adaption to attenuate the damage in zebrafish model that have potential to resist hypoxic environment.METHODS The hypoxic zebrafish model was established in different concentration of oxygen with 3%,5%,10%,21%in water.The brain tissue was separated and the RNA-seq was used to identify the differentially expressed genes.The related endogenous metabolites profiles were obtained by LC-HDMS,and the multivariate statistics was applied to discover the important metabolites candidates in hypoxic zebrafish.The candidates were searched in HMDB,KEGG and Lipid Maps databases.RESULTS The zebrafish hypoxic model was successfully constructed via the different concentration of oxygen,temperature and hypoxic time.The activities of the related hypoxic metabolic enzymes and factors including HIF-1a,actate dehydrogenase(LDH)and citrate synthase(CS)were evaluated.Significant differences(P<0.05 and fold change>2)in the expression of 422 genes were observed between the normal and 3% hypoxic model.Among them,201 genes increased depended on the lower concentration of oxygen.53 metabolites were identified that had significant difference between the hypoxia and control groups(P<0.05,fold change>1.5 and VIP>1.5).The ten key metabolites were increased gradually while six compounds were decreased.The endogenous hypoxic metabolites of phenylalanine,D-glucosamine-6P and several important lipids with the relevant hub genes had similar change in hypoxic model.In addition,the metabolic pathways of phenylalanine,glutamine and glycolipid were influenced in both the levels of genes and metabolites.CONCLUSION The up-regulation of phenylalanine,D-glucosamine-6P and lipid may have further understanding of protective effect in hypoxia.Our data provided an insight to further reveal the hypoxia and adaptation mechanism.

Establishment of an adult zebrafish model of retinal neurodegeneration induced by NMDA

AIM: To establish a model of retinal neurodegeneration induced by N-Methyl-D-aspartic acid(NMDA) in adult zebrafish.METHODS: We compared the effects of three different NMDA delivery methods on retinal neurodegeneration in adult zebrafish: immersion(I.M.), intravitreal injection(I.V.), and intraperitoneal injection(I.P.), and examined retinal pathology and degeneration by hematoxylin and eosin and TUNEL staining in the treated zebrafish. Effects of the NMDA receptor antagonist MK-801 and the natural product resveratrol on NMDA-induced retinal neurodegeneration were also assessed.RESULTS: The thickened inner retina was seen in histology with 100 μmol/L NMDA by I.M. administration. Significant apoptosis in the retinal ganglion cell layer and retinal thickness reduction occurred in 0.5 mol/L NMDA I.P. administration group.Seizure-like behavioral changes, but no retinal histological alteration occurred in 16 mg/kg NMDA I.P. administration group. Resveratrol and MK-801 prevented NMDA-induced retinal neurodegeneration in the zebrafish. CONCLUSION: Among the three drug administration methods, I.V. injection of NMDA is the most suitable for establishment of an acute retinal damage model inzebrafish. I.M. with NMDA is likely the best for use as a chronic retinal damage model. I.P. treatment with NMDA causes brain damage. Resveratrol and MK801 may be a clinically valuable treatment for retinal neurodegeneration.

c-myb is involved in CML progression and is a therapeutic target in the zebrafish CML model

Background:Despite the success of tyrosine kinase inhibitors in chronic myeloid leukemia(CML)therapy,CML still faces the challenges of drug resistance and progression to blast crisis.Twenty-five percent of patients have imatinib resistance and treatment difficulties due to heterogeneity after progression,but little is known about the mechanism.A key transcription factor in hematopoiesis,MYB,has been reported to increase abnormally in several types of aggressive blood disorders including CML.Methods:This study used a zebrafish model to explore the relationship between BCR/ABL1 and c-myb in CML progression.A CML zebrafish model was crossed with a c-myb hyperactivity transgenic line.Results:It was found that both exogenous BCR/ABL1 and c-myb could up-regulate the expression of neutrophil-related genes.More seriously,neutrophil accumulation was observed when BCR/ABL1 was combined with c-myb overexpression.Further studies showed that c-myb may be one of the downstream targets of BCR/ABL1 and the effect of BCR/ABL1 on neutrophils was c-myb dependent.Taking advantage of this inheritable in vivo model,it was shown that a combination of imatinib and flavopiridol,a cyclin-dependent kinase inhibitor targeting MYB,could more effectively alleviate the aggressive phenotype of the double transgene line.Conclusion:In summary,this study suggests that c-myb acts downstream of BCR/ABL1 and is involved in CML progression and is therefore a risk factor and a valuable target for the treatment of CML progression.The model used in the study could be helpful in high-throughput drug screening in CML transformation.

A novel zebrafish vascular injury model for asseessing drug efficancy of Yangxue Qingnao granules

OBJECTIVE To assess the efficancy of Yangxue Qingnao granules on simvastatininduced vascular injury model in zebrafish.METHODS Since statins can inhibit vascular development in zebrafish,in this study,we developed a novel animal model using 1 to 3 day post-fertilization larval zebrafish by optimizing the doses and duration of simvastatin exposure.Five pro-angiogenic drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model.Zebrafish was treated with different concentration of Yangxue Qingnao granules(62.5,125 and 250 μg·mL-1) for 2 d then tested for the area of subintestinal vein vessels.RESULTS Vascular regeneration promoting effect of five pro-angiogenic drugs(calycosin,astragaloside,chlorogenic acid,ferulic acid and Panax Notoginseng Saponins) were 8-48%,24-51%,35-58%,28-75% and 37-69%,respectively.In 62.5,125 and 250 μg·mL-1 Yangxue Qingnao granules group,vascular regeneration promoting effect were 21%(P>0.05),84%(P<0.01) and 53%(P<0.01).CONCLUSION Our results demonstrate that the zebrafish vascular injury model validated in this study could be used for in vivo angiogenesis studies and drug screening and for assessing pro-angiogenic drugs with different mechanisms.Yangxue Qingnao granules could promote the vascular regeneration in zebrafish.

Comparison of Two Chemically-Induced Colitis-Models in Adult Zebrafish, Using Optical Projection Tomography and Novel Transcriptional Markers

Crohn’s disease and ulcerative colitis—inflammatory bowel diseases (IBD)—are chronic conditions with an inadequately understood pathogenesis. Employing a set of novel molecular markers in a gene expression assay (qPCR), we have used adult zebrafish to investigate two acute inflammatory models, induced by the haptenizing agents oxazolone and TNBS. In addition, goblet cell dynamics in the scales and intestine and 5-HT (serotonin) in intestinal tissues were investigated through optical projection tomography. Gene expression studies revealed a distinct and significant upregulation of proinflammatory cytokines, acute-phase reactants and metalloprotease 9 in both chemical models, primarily after 72 hours. In comparison, transcription factors and cytokines associated with Th1 and Th17 (Crohn’s) and Th2 (ulcerative colitis) were mainly not affected in this acute setting. However, elevated transcript levels were detected in Foxp3, IL-10 and T-bet, which are linked with tolerance and Tregs in mammals. Goblet cells in scales were depleted in both chemical models and in the intestine of oxazolone-treated fish. A marked 5-HT signal was noted in intestinal tissue of some chemically treated zebrafish. In conclusion, a distinct acute inflammatory reaction was induced in both chemical models. Further, oxazolone and TNBS did not result in clear-cut Th2 and Th1/Th17 pathway signaling at this early timepoint, but the applied molecular tools may provide further insight to the IBD pathogenesis and translational value of the IBD zebrafish model.

斑马鱼模型在中医药防治骨质疏松应用的研究进展

随着骨质疏松(osteoporosis,OP)动物模型的不断发展,斑马鱼逐渐成为中医药防治OP的常用动物模型之一。文章整理了近五年来以斑马鱼作为动物模型在OP防治方面的运用,系统综述了不同种类斑马鱼OP模型的建立以及通过斑马鱼对单体中药、中药复方以及中成药对OP防治的药理、毒理的研究进展。以期对OP发病机制以及抗OP药物筛选、研究的动物模型选择提供新的思路与方法。

Animal models of amyotrophic lateral sclerosis： a comparison of model validity

Animal models are necessary to investigate the pathogenic features underlying motor neuron degeneration and for therapeutic development in amyotrophic lateral sclerosis（ALS）. Measures of model validity allow for a critical interpretation of results from each model and caution from over-interpretation of experimental models. Face and construct validity refer to the similarity in phenotype and the proposed causal factor to the human disease, respectively. More recently developed models are restricted by limited phenotype characterization, yet new models hold promise for novel disease insights, thus highlighting their importance. In this article, we evaluate the features of face and construct validity of our new zebrafish model of environmentally-induced motor neuron degeneration and discuss this in the context of current environmental and genetic ALS models, including C9 orf72, mutant Cu/Zn superoxide dismutase 1 and TAR DNA-binding protein 43 mouse and zebrafish models. In this mini-review, we discuss the pros and cons to validity criteria in each model. Our zebrafish model of environmentally-induced motor neuron degeneration displays convincing features of face validity with many hallmarks of ALS-like features, and weakness in construct validity. However, the value of this model may lie in its potential to be more representative of the pathogenic features underlying sporadic ALS cases, where environmental factors may be more likely to be involved in disease etiology than single dominant gene mutations. It may be necessary to compare findings between different strains and species modeling specific genes or environmental factors to confirm findings from ALS animal models and tease out arbitrary strain-and overexpression-specific effects.

Housing Environment Affects Baseline Anxiety in Zebrafish

Neuroscience researchers that wish to address compelling questions in psychopharmacology would benefit from the use of zebrafish, which have a behavioral repertoire that is rich and complex and that reflects many fundamental processes in humans, such as those that evoke anxiety. Zebrafish behavior is easily quantified in diverse test environments, but it is important to consider the conditions used to house the fish, prior to the behavioral testing, when designing experiments. Studies show that fish housed at high densities exhibit elevated cortisol levels as compared with fish housed at lower densities. The literature is less clear about the behavioral consequences of housing and handling. We examined the effects of housing (group versus paired) on several behavioral parameters. We observed that group-housed fish exhibited an anxious phenotype as evidenced by tank diving and meandering. Other more general indicators of movement and spatial navigation were unaffected by housing. This study focused on two specific housing environments, and the results support the need for continued research in this area to further elucidate the contributions of housing and handling on the subtleties of zebrafish behavioral markers.

Novel MIP gene mutation causes autosomal-dominant congenital cataract

●AIM:To identify disease-causative mutations in families with congenital cataract.●METHODS:Two Chinese families with autosomaldominant congenital cataract(ADCC)were recruited and underwent comprehensive eye examinations.Gene panel next-generation sequencing of common pathogenic genes of congenital cataract was performed in the proband of each family.Sanger sequencing was used to valid the candidate gene mutations and sequence the other family members for co-segregation analysis.The effect of sequence changes on protein structure and function was predicted through bioinformatics analysis.Major intrinsic protein(MIP)-wildtype and MIP-G29R plasmids were constructed and microinjected into zebrafish single-cell stage embryos.Zebrafish embryonic lens phenotypes were screened using confocal microscopy.●RESULTS:A novel heterozygous mutation(c.85G>A;p.G29R)in the MIP gene was identified in the proband of one family.A known heterozygous mutation(c.97C>T;p.R33C;rs864309693)in MIP was found in the proband of another family.In-silico prediction indicated that the novel mutation might affect the MIP protein function.Zebrafish embryonic lens was uniformly transparent in both wild-type PCS2+MIP and mutant PCS2+MIP.●CONCLUSION:Two missense mutations in the MIP gene in Chinese cataract families are identified,and one of which is novel.These findings expand the genetic spectrum of MIP mutations associated with cataracts.The functional studies suggest that the novel MIP mutation might not be a gain-of-function but a loss-of-function mutation.

基于斑马鱼模型研究康达心口服液对阿霉素心脏毒性的预防和作用机制

目的基于斑马鱼模型探究复方中药康达心口服液对阿霉素(doxorubicin,DOX)所致心脏毒性的预防及作用机制。方法将实验对象分为空白对照组、阿霉素造模组、康达心低浓度组、康达心中浓度组和康达心高浓度组。设计阿霉素和康达心浓度梯度,筛选出35μg/mL的阿霉素作为诱导浓度,5.9325×10^(-2)μg/mL、11.875×10^(-2)μg/mL和23.75×10^(-2)μg/mL的康达心分别作为低浓度、中浓度和高浓度治疗浓度。显微镜观察斑马鱼的畸形和心率等情况;TUNEL荧光观察心脏细胞的凋亡情况;Q-PCR和Western blot检测心脏发育相关基因及蛋白(SOX9b、GATA4、NKX2.5)的表达和凋亡相关基因及蛋白(P53、Caspase-9、bcl2、Bax、TNF-α)的表达。结果与空白对照组比较,阿霉素造模组和康达心低浓度组斑马鱼心率显著下降(P<0.05),而康达心中浓度组和高浓度组斑马鱼心率显著升高(P<0.05);荧光染色显示,与空白对照组比较,阿霉素造模组心脏细胞凋亡升高(P<0.05),不同浓度康达心共处理组心脏细胞凋亡显著降低(P<0.05);Q-PCR和Western blot检测显示,与空白对照组比较,阿霉素造模组心脏发育相关基因及蛋白表达、凋亡相关基因表达显著下降(P<0.05);与阿霉素造模组比较,不同浓度康达心处理组使心脏发育相关基因及蛋白表达、凋亡相关基因表达升高(P<0.05)。结论康达心口服液对阿霉素诱导的心脏毒性具有显著的预防作用,对心肌具有保护作用,也为其临床应用提供了科学依据。

Efficacy Assessment of Whitening Cosmetics in Animal Models

Animal skin is generally preferred to evaluate the efficacy and safety:several animal models have been used,e.g.,the guinea pig,the mouse,and the zebrafish.The epidermis of guinea pigs displays a moderate number of melanocytes and melanosomes distributed in a similar way to human skin.Guinea pig and human skins have given the close morphologic and functional similarities(similar epidermis thickness,similar epidermal cells turnover time,etc.).The zebrafish presents several advantages,including easy maintenance and handling of the animals,short generation times,and high efficiency of drug penetration through the skin.To establish the animal model and to assess whitening efficacy and safety for whitening products.

基于模式生物斑马鱼对口腔炎合剂的胚胎及肝肾发育的影响

目的利用斑马鱼为动物模型,初步分析口腔炎合剂对胚胎及肝肾发育的影响。方法待斑马鱼发育至受精后10 h(10 hpf)时以简单随机方法分组并添加梯度浓度的口腔炎合剂。药物作用72 h后在显微镜下观察口腔炎合剂对胚胎及肝肾发育,并记录图像数据。结果当口腔炎合剂稀释浓度高于1.625×10^(-5)g/mL时对10 hpf斑马鱼开始出现胚胎发育毒性,表现为发育迟缓、心腔狭窄等。当口腔炎合剂稀释浓度高于6.500×10^(-5)g/mL时对36 hpf斑马鱼产生肝肾发育毒性,主要表现为肝脏与周边组织分界线不太明显,肾脏和鱼鳔区域明显增大,并伴有水肿瘀滞现象。结论口腔炎合剂具有一定的胚胎及肝肾毒性,孕妇和哺乳期妇女,以及肝肾功能不全者原则上不得使用本品,必须使用时应严格限量,并在医生指导下谨慎使用。

核桃粕源抗氧化活性肽的酶解制备及活性分析

目的:深度利用核桃粕蛋白。方法:采用分级分离法从核桃粕中提取蛋白质后,分别用胰蛋白酶、木瓜蛋白酶、中性蛋白酶、碱性蛋白酶和风味蛋白酶5种蛋白酶对其进行水解,并比较分析胰蛋白酶酶解物(trypsin hydrolysate,TH)、木瓜蛋白酶酶解物(papain hydrolysate,PH)、中性蛋白酶酶解物(dispase hydrolysate,DH)、碱性蛋白酶酶解物(alcalase hydrolysate,AH)和风味蛋白酶酶解物(flavourzyme hydrolysate,FPH)的抗氧化活性;以DPPH自由基清除率、ABTS自由基清除率和羟自由基清除率为检测指标分析不同浓度AH的体外抗氧化性;采用LC-MS/MS技术鉴定AH中所有肽的序列,并利用PeptideRanker、BIOPEP-UWM数据库和分子对接工具AutoDock1.5.6筛选潜在的核桃抗氧化活性肽;将上述两条肽分别与Keap1蛋白进行计算机模拟分子对接,并利用斑马鱼胚胎氧化损伤模型,对人工合成的两条抗氧化活性肽的抗氧化活性进行分析。结果:5种酶解物中抗氧化活性较好的AH对DPPH自由基、ABTS自由基和羟自由基的最佳清除率分别为(71.56±0.75)%,(60.63±0.45)%,(98.63±0.06)%。LC-MS/MS鉴定并通过计算机分析预测得到两条活性最优的肽,即:ALWPF和PLRWPF;两条抗氧化活性肽均能与Keap1蛋白的活性部位结合,其结合能分别为-9.2,-9.6 kJ/mol。ALWPF和PLRWPF肽处理斑马鱼胚胎的安全质量浓度范围分别为0~50,0~30μg/mL。结论:从核桃粕中鉴定出两条抗氧化肽,其对斑马鱼胚胎具有保护作用。

光照、咖啡因及联合法诱导建立斑马鱼失眠模型的比较研究

目的利用斑马鱼建立不同睡眠剥夺模型,为失眠的基础研究提供更多可重复性的实用建模参考方案。方法用持续光照(150 Lux)和光照加咖啡因两种干预手段诱导斑马鱼失眠模型,随机将斑马鱼分为空白组、光照组、咖啡因组(100μmol/L)和联合组。利用旷场实验和昼夜节律行为学实验观察各组斑马鱼运动能力的区别;利用qPCR技术与ELISA技术检测相关睡眠基因与5-HT神经递质的表达与分泌情况。结果与空白组相比,光照组斑马鱼睡眠时间、静息时间(白天)、睡眠回合明显减少(P<0.01或P=0.01),移动距离较空白组明显增加(P<0.01)。联合组与咖啡因组斑马鱼静息时间(白天)、睡眠回合较空白组增加(P<0.01)。联合组与咖啡因组斑马鱼活动距离无统计学意义(P>0.05);与空白组相比,光照组中心区域游动距离百分比与中心区域游动时间百分比均降低(P<0.05);与光照组相比,咖啡因组中心区域游动距离百分比与中心区域游动时间百分比均降低(P<0.01);与空白组相比,6:00与12:00光照组斑马鱼HTR1aa mRNA表达呈上调趋势(P<0.05),光照组与联合组斑马鱼体内HTR1ab mRNA水平表达均无明显统计学意义(P>0.05);与空白组相比,6:00时光照组的5-HT分泌量均减少(P<0.01);12:00光照组与联合组5-HT量均减少(P<0.01);18:00时光照组与联合组分泌量依然低于空白组(P<0.01)。结论单纯光照法最适用于斑马鱼幼鱼长效失眠模型的建立,其相关机制可能与HTR1aa基因以及5-HT等生物因子有关。

斑马鱼模型在特发性脊柱侧凸研究中的应用

特发性脊柱侧凸(IS)指原因不明的脊柱发育异常,给患者带来身体和经济的双重负担。然而,目前IS的致病机制尚未完全明确,疾病的治疗通常以大创口的外科手术干预为主。为了探索IS的病因,并为新的诊疗方法提供理论基础,采用各种模式动物进行实验尤为必要。作为一种新兴的模式动物,斑马鱼繁殖周期短、培育成本低,且具备体外受精、体外发育和胚胎透明等特点,因此其遗传编辑和观测均较传统模式动物更容易。目前斑马鱼模型已成为研究IS的重要工具,其在揭示IS的分子机制和潜在治疗靶点方面展现出巨大潜力。

斑马鱼在肝病模型中的研究进展

肝脏是人体进行能量代谢的主要场所。肝病是严重影响人们生活质量的临床疾病之一,多种因素导致肝脏功能及结构异常。目前关于肝病模型的基础研究已取得一定进展,多种肝病模型用于肝病机制的研究,可有效复制肝病的临床特征及病变过程。采用大鼠、小鼠复制肝病模型均存在弊端,基础实验研究中采用斑马鱼复制肝病模型具有自身优势,在肝病治疗药物筛查中具有重要意义。全面了解斑马鱼在肝病模型中的作用,可以为相关疾病的治疗提供新思路。

棕榈酸诱导斑马鱼脂毒性损伤骨形成抑制模型的建立

目的 建立棕榈酸诱导的斑马鱼脂毒性损伤骨形成抑制模型。方法 将AB品系斑马鱼胚胎分为对照组、棕榈酸组(PA组)和辛伐他汀组(SIM组)。从3 dpf(days post fertilization, dpf)开始,PA组、SIM组给予PA造模。从5 dpf开始,SIM组给予SIM连续给药4 d。9 dpf通过钙黄绿素染色法、尼罗红染色法、甘油三酯及总胆固醇含量测定、q-PCR判断模型是否成功建立。结果 PA显著减少斑马鱼椎骨骨节数目、促进脂质堆积、增加甘油三酯及总胆固醇含量、促进成脂相关基因PPARγ、c/EBPα的表达并抑制成骨相关基因ALP、RUNX2的表达。SIM可以改善PA对斑马鱼的骨形成抑制效应。结论 采用PA给药的方法可成功造成与骨质疏松症(osteoporosis, OP)病理过程相似的脂毒性损伤骨形成抑制模型,该方法简便、灵敏、可控,可用于OP及相关疾病的药物筛选。

利用斑马鱼肝癌模型探究氨基葡萄糖盐酸盐对肝癌的影响

目的:探究双孢蘑菇来源的氨基葡萄糖盐酸盐(Glucosamine Hydrochloride,GAH)对肝癌的影响。方法:利用斑马鱼肝癌及血管双转基因模型Tg(flk:mCherry;kras^(G12V)),在60 mg/mL盐酸多西环素溶液的诱导下,通过激光共聚焦显微镜追踪观察第5 d到第10 d肝癌的发生发展;在Dox诱导的同时加入不同浓度GAH恢复,在激光共聚焦显微镜下比较分析正常组(Ctr)、模型组(Doxorubicin,Dox)、Dox+0.1%GAH组、Dox+0.3%GAH组中GAH对肝癌及肝癌血管生长的影响;通过qPCR和ELISA实验分析GAH对肝癌及肝癌血管相关调控因子mRNA和蛋白水平表达的影响(prmt5、tiam1、kat5、vegfaa、vegfr2、tgf-β1);最后通过TIMER2在线生物信息学分析验证tgf-β1与prmt5、tiam1、kat5与vegfaa、vegfr2的相关性。结果:Dox持续诱导会引起肝癌的发生发展;与Dox模型组相比,GAH处理极显著抑制肝癌的生长与侵袭(P<0.001),并能极显著改善肝癌血管紊乱(P<0.001);qPCR和ELISA实验结果表明GAH能够显著抑制肝癌及血管相关基因的mRNA和蛋白水平(P<0.05);生物信息学分析结果提示tgf-β1与肝癌及血管相关基因的表达具有明显相关性。结论:GAH可能通过调控TGF-β信号通路影响肝癌相关基因的表达抑制肝癌血管的生成,从而抑制肿瘤的生长与侵袭。

发酵麸皮多糖对胆固醇诱导的高脂斑马鱼脂肪代谢和抗氧化相关指标的影响

试验旨在研究发酵麸皮多糖(FWBP)对胆固醇诱导的高脂斑马鱼脂肪代谢和抗氧化相关指标的影响。试验主要采用5 dpf野生型AB斑马鱼作为研究对象,先构建高脂斑马鱼模型,明确5%的高胆固醇饲料可以引起斑马鱼体内脂肪积累显著增加,在此基础上分别添加(0、0.25‰、0.50‰、0.75‰、1.00‰)的FWBP,研究FWBP是否可以改善胆固醇引起的脂肪代谢和抗氧化功能异常,每组5个重复,每个重复30尾仔鱼,试验期10 d。结果表明:与对照组相比,0.50‰FWBP组斑马鱼体内总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白(LDL)含量,以及谷草转氨酶(AST)、谷丙转氨酶(ALT)和谷氨酰转肽酶(GGT)活性均显著降低(P<0.05);1.00‰FWBP组鱼体高密度脂蛋白(HDL)含量均显著降低(P<0.05)。与对照组相比,0.25‰FWBP组、0.50‰FWBP组、0.75‰FWBP组鱼体超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性均显著提高(P<0.05),各试验组的丙二醛(MDA)含量显著降低(P<0.05)。研究表明,FWBP可以有效降低摄入高脂饲料后斑马鱼体总脂肪和脂蛋白的含量,降低体内脂肪代谢相关酶活性和总胆红素(TBIL)的含量,提高抗氧化酶活性,降低MDA含量,缓解高脂引起的氧化应激,添加0.50‰时效果较好。