Metabolic dysfunction-associated fatty liver disease (MAFLD) is a prevalent liver disease that poses a serious health threat. Fisetin (FIS) is a flavonoid compound with multiple biological activities. The aim of this study was to investigate the potential therapeutic effect of FIS on MAFLD and the underlying mechanisms through in vivo and in vitro experiments. Our results demonstrated that FIS mitigated weight gain and improved blood glucose levels, blood lipid disorder and liver steatosis in mice with high-fat diet (HFD)-induced MAFLD. RNA-seq analysis revealed that FIS affected the expression of many genes in the liver of MAFLD mice, including up-regulating the mRNA expression of Glut4 and GPx3 and down-regulating the mRNA expression of IL-1β and Cxcl10 genes. Moreover, FIS reduced sodium oleate-induced lipid accumulation, activated the GSK-3β/Nrf2/HO-1 signaling pathway, and inhibited the expression of PEPCK and G6PC in HepG2 cells. Overall, these findings demonstrate that FIS can alleviate MAFLD by enhancing the antioxidant capacity and reducing the gluconeogenic capacity in the liver, supporting FIS as a potential therapeutic against MAFLD.

The objective of this research was to examine the protective effects of ellagic acid (EA) against diabetes-induced atherosclerosis in rats and oxidative damage in vascular endothelial cells. In Type 2 diabetic rats induced by high-fat diet and streptozotocin (STZ), a four-week EA treatment noticeably reduced serum lipid deposition, improved oxidative stress indicators, alleviated aortic endothelial atherosclerosis, and restored the diversity and the composition of the gut microbiota. Correlation analysis demonstrated significant associations among oxidative damage markers and gut microbiota influenced by EA. RNA sequencing in EA-treated human umbilical vein endothelial cells (HUVECs) and network pharmacology identified NAD(P)H dehydrogenase quinone 1 (NQO1), heme oxygenase-1 (HO-1), and superoxide dismutase-2 (SOD2) as potential key antioxidant genes. Furthermore, experiments in a high-glucose or hydrogen peroxide (H2O2)-induced oxidative damage model in HUVECs confirmed that EA significantly enhanced cellular vitality, reduced cellular oxidative damage, and upregulated NQO1, HO-1, and SOD2 expression at both gene and protein levels. In summary, these findings demonstrate that EA mitigates diabetic atherosclerosis by enhancing endothelial antioxidant capacity and modulating gut microbiota.

Objective Since the liver is key to poultry lipid metabolism and fasting models are widely used in studying animal nutrition metabolism, this study used fasting to explore chicken liver lipid metabolism characteristics, providing a basis for poultry lipid metabolism research. Methods RNA-seq and metabolomics were combined to analyze 72-hour fasting effects on chicken liver lipid metabolism. Key lipid metabolism-related genes and metabolites were identified, with their mechanisms explored via RNAi and Oil Red O staining. Results Metabolomics identified 648 differential metabolites, including 8 (e.g., Arachidonoyl amine) with levels 160-fold higher than controls. Transcriptomics found 849 differentially expressed genes (DEGs), 78 involved in lipid metabolism; Protein–protein interaction analysis revealed hub genes like EHHADH. Transcriptome-metabolome correlation analysis showed 101 DEGs correlated with 99 metabolites, with EHHADH associated with 54 metabolites (the most) and linked to 2-methylcrotonoyl-CoA and 5 pathways via KEGG Markup Language. Fasting upregulated EHHADH, whose overexpression/interference altered mRNA levels of Fabp7, Plin2, ACACA, FASN, PPARα, as well as cholesterol/triglyceride levels. EHHADH overexpression reduced LMH cell lipid deposition, while interference increased it, indicating its role in reducing lipid deposition. Conclusion Fasting significantly alters chicken liver lipid metabolism, and EHHADH helps reduce liver lipid deposition.

Geraniin (GE), an ellagic tannin extracted from Geranium wilfordii, has been demonstrated to improve high-fat diet-induced metabolic disorders. This study aimed to investigate the effect of Geraniin on atherosclerosis and the underlying mechanism. In this study, we used the ApoE -/- mice as a model of atherosclerosis. We fed the ApoE -/- mice a high-fat diet containing 1.25% cholesterol and treated them with or without 5 or 10 mg/kg body weight of GE for 10 weeks. Meanwhile, we combined network pharmacology analysis to explore how GE improves H2O2-induced endothelial cell apoptosis and inflammatory response. Our results showed that GE treatment significantly reduced serum concentrations of lipids, oxidative stress damage, lipid deposition, and atherosclerotic plaque lesions in the ApoE -/- mice. Furthermore, a network pharmacology analysis indicated that the beneficial effect of GE treatment on atherosclerosis is closely related to its effect on oxidative stress, cell apoptosis, and inflammatory responses. We also showed that in H2O2-treated HUVEC cells, GE promoted the release of NO, increased the activity of antioxidant enzymes, reduced endothelial cell apoptosis, reduced the production of inflammatory cytokines IL1β, IL6, and TNFα, and activated the Akt/eNOS/NO and GSK-3β/Nrf2/HO-1 signaling pathways. Taken together, these findings demonstrate that GE can alleviate high-fat diet-induced atherosclerosis in ApoE -/- mice by inhibiting oxidative stress-induced endothelial cell apoptosis and inflammation.

OBJECTIVE:Vanin-1 (VNN1) is a pantetheinase that catalyses the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies have shown that the VNN1 is specifically expressed in chicken liver which negatively regulated by microRNA-122. However, the functions of the VNN1 in lipid metabolism in chicken liver haven't been elucidated.METHODS:First, we detected the VNN1 mRNA expression in 4-week chickens which were fasted 24 hours. Next, knocked out VNN1 via CRISPR/Cas9 system in the chicken Leghorn Male Hepatoma cell line. Detected the lipid deposition via oil red staining and analysis the content of triglycerides (TG), low-density lipoprotein-C (LDL-C), and highdensity lipoprotein-C (HDL-C) after VNN1 knockout in Leghorn Male Hepatoma cell line. Then we captured various differentially expressed genes (DEGs) between VNN1-modified LMH cells and original LMH cells by RNA-seq.RESULTS:Firstly, fasting-induced expression of VNN1. Meanwhile, we successfully used the CRISPR/Cas9 system to achieve targeted mutations of the VNN1 in the chicken LMH cell line. Moreover, the expression level of VNN1 mRNA in LMH-KO-VNN1 cells decreased compared with that in the wild-type LMH cells (p<0.0001). Compared with control, lipid deposition was decreased after knockout VNN1 via oil red staining, meanwhile, the contents of TG and LDL-C were significantly reduced, and the content of HDL-C was increased in LMH-KO-VNN1 cells. Transcriptome sequencing showed that there were 1,335 DEGs between LMH-KO-VNN1 cells and original LMH cells. Of these DEGs, 431 were upregulated, and 904 were downregulated. Gene ontology analyses of all DEGs showed that the lipid metabolism-related pathways, such as fatty acid biosynthesis and long-chain fatty acid biosynthesis, were enriched. KEGG pathway analyses showed that "lipid metabolism pathway", "energy metabolism", and "carbohydrate metabolism" were enriched. A total of 76 DEGs were involved in these pathways, of which 29 genes were upregulated (such as cytochrome P450 family 7 subfamily A member 1, ELOVL fatty acid elongase 2, and apolipoprotein A4) and 47 genes were downregulated (such as phosphoenolpyruvate carboxykinase 1) by VNN1 knockout in the LMH cells.CONCLUSION:These results suggest that VNN1 plays an important role in coordinating lipid metabolism in the chicken liver.

鞣花酸是一种多酚物质,广泛存在于多种植物中,具有抗氧化、抗肿瘤、抗菌、抗炎、抗病毒、保护肝脏及心血管等多种药理作用.本文在查阅近二十年文献的基础上对鞣花酸药理作用进行综述,以期进一步开发鞣花酸在临床上的应用.

肝核因子3β(hepatocyte nuclear factor 3β,HNF 3β)作为一个重要的转录调控因子,参与哺乳动物的前肠胚和肝脏发育以及成体肝内的能量代谢,而HNF 3β在鸡(Gallus gallus)中的功能还不清楚.为研究鸡HNF 3β在肝内糖脂代谢中的作用提供有效的抗体,本研究优化合成了鸡Hnf 3β基因编码区片段,以pET 30a表达载体在大肠杆菌(Escherichia coli)中进行重组蛋白HNF 3β的表达,用6 mol/L尿素变性包涵体,Ni2+对重组蛋白进行亲和层析,并以快速稀释法复性重组蛋白,以离心超滤法收集、浓缩目的蛋白;取纯化的HNF 3β重组蛋白和弗氏佐剂乳化后免疫兔(Oryctolagus cuniculus),制备鸡HNF 3β的抗血清,以酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)测定抗血清效价后,用Protein A法纯化抗体;通过检测鸡肝癌细胞系(leghorn male hepatoma,LMH)中HNF 3β蛋白的表达情况验证所制备抗体的效果.结果表明,优化合成的Hnf 3β基因片段在大肠杆菌中实现了重组蛋白的高效表达;在6 mol/L尿素的变性条件下,Ni2+亲和层析一步法获得纯度大于95％的重组蛋白,结合快速稀释法获得了可溶性的重组蛋白HNF 3β;经重组蛋白4次免疫后,抗血清效价超过1:64000;免疫印迹结果显示,制备的抗体能特异性的检测出LMH细胞中HNF 3β蛋白的表达.综上所述,本研究利用制备的高纯度重组蛋白HNF 3β免疫兔获得了高特异性和高效价的多克隆抗体,可以为检测鸡HNF 3β蛋白分子及研究其功能提供可靠的抗体.

Nonalcoholic fatty liver disease (NAFLD) is the most common liver metabolic disease in the world. In this study we investigated the effect of tetrahydrocurcumin (THC), a potent antioxidant, on NAFLD. We found that THC significantly reduced the body weight, serum and liver lipids, serum malondialdehyde (MDA) and fasting blood glucose (FBG) of male NAFLD mice induced by feeding a high-fat diet (HFD) for 8 weeks. A RNA-seq analysis revealed that THC also affected the expression of many genes involved in lipid metabolism in the liver of NAFLD mice, including increasing the mRNA expression of CYP51 and FOXQ1. Moreover, THC significantly reduced sodium oleate-induced lipid accumulation, activated the NRF2 pathway, and up-regulated the mRNA expression of FGF21 in HepG2 cells. Overall, our study showed that THC can improve the antioxidant capacity of liver, which suggests that dietary THC may be used to treat NAFLD.

为探究Elovl2基因在鸡体内的生物学功能,对鸡ELOVL2蛋白的理化性质进行分析,通过RT-qPCR技术检测该基因在鸡各组织中的表达情况.将Elovl2基因过表达重组质粒与最佳干扰片段分别转染鸡LMH细胞系,通过RT-qPCR技术检测Elovl2基因及脂代谢相关基因mRNA水平的变化,利用GC-MS技术检测细胞内相关脂肪酸含量的变化.结果显示:鸡ELOVL2蛋白属于稳定蛋白,该基因mRNA主要表达于脑和肝脏组织.转染pcDNA3.1-Elovl2后,基因相对表达水平显著上升,过表达重组质粒构建成功.三组siRNA均显著抑制Elovl2基因表达,抑制效果最佳的为siRNA-681.抑制Elovl2基因表达后,pck1基因的表达量较对照组相比上升2倍,但二者差异不显著.过表达Elovl2基因后,各种脂肪酸含量均有所增加,而经RNAi技术抑制Elovl2基因表达后,各种脂肪酸则有不同程度减少,受其影响最明显的为C22:1N9.试验结果为进一步研究鸡Elovl2基因的功能奠定了基础.

鸡肉占肉质消费品首位,过多的脂质沉积会降低肉质.研究表明自噬参与肝脏脂质代谢,改善脂质沉积,从而调控禽类脂质代谢.本研究旨在探究自噬相关基因5(autophagy related gene 5,Atg5)对鸡(Gallus gallus)成纤维细胞系DF-1细胞内的脂质代谢相关基因的影响.根据NCBI数据库中鸡ATG5蛋白(NP_001006409.1)氨基酸序列,利用ExPASy数据库对鸡ATG5蛋白进行理化特性分析;利用qPCR技术检测鸡各组织中Atg5基因的表达情况;构建过表达重组质粒pcDNA3.1-Atg5,并且利用RNAi技术干扰Atg5表达;最后,检测干扰和过表达Atg5后相关脂代谢基因:载脂蛋白B(apolipoprotein,ApoB)、脂肪酸合成酶(fatty acid synthase,FASN)、乙酰辅酶A羧化酶(acetyl CoA carboxylase,ACACA)、过氧化物酶体增殖物激活受体-α(peroxisome proliferator-activated receptor-α,PPARα)、叉头转录因子-1(forkhead box protein-1,Foxo1)、脂肪酸结合蛋白7(fatty acid-binding protein 7,FABP7)、分拣蛋白1(sortilin 1,Sort1)的表达情况;结果表明:鸡ATG5蛋白是相对分子质量为32.478 kD的不稳定蛋白质,Atg5在各组织中均有表达,其中以胸肌组织中表达量最高;此外,成功构建过表达载体pcDNA3.1-Atg5,筛选出最优干扰siRNA—siRNA-330,过表达或干扰Atg5基因对其他脂代谢相关基因的表达情况均有影响;其中,过表达Atg5后,ApoB的表达量显著降低,Sort1的表达量显著增加,FABP7变化不显著;干扰Atg5后,FABP7和ApoB的表达量显著提高,Sort1变化不显著,推测三者与Atg5可能存在潜在联系.本研究为进一步研究鸡Atg5基因的功能提供参考依据.

旨在探究Apob基因在鸡肝脂质代谢过程中的功能。本研究对鸡Apob蛋白进行理化性质分析；利用RT-qPCR检测Apob基因在4周龄黄羽肉鸡组织中的表达情况，每组设置3个重复，进行3次平行试验。根据鸡Apob蛋白关键结构域，在Apob基因外显子设计3对sgRNA，构建Cas/gRNA载体；将重组质粒转染DF-1细胞后，利用T7核酸内切酶Ⅰ（T7 endonuclease Ⅰ，T7EI）酶切法和TA克隆测序法筛选敲除活性位点并计算敲除效率。利用RT-qPCR检测基因敲除后亚克隆细胞中Apob基因mRNA表达情况。结果表明，鸡Apob的相对分子质量为523.356 ku，平均亲水性为-0.300，为稳定的蛋白质，并且该基因主要在鸡的肝、肾和小肠组织表达。敲除载体转染至DF-1细胞后，T7EI酶切发现，Cas/gRNA6、Cas/gRNA7和Cas/gRNA8三个位点均可发挥敲除活性，TA克隆测序结果表明，三者的敲除效率分别为33.3%、65%和80%。同时，RT-qPCR结果显示，转染Cas9/gRNA7、Cas9/gRNA6、Cas9/gRNA8的细胞中Apob基因mRNA表达水平约分别下调99.96%（P < 0.01）、85%（P < 0.01）、47%（P < 0.05）。综上所述，本研究揭示了鸡Apob基因在组织中的表达特点和蛋白的理化性质；成功构建了鸡Apob基因CRISPR/Cas9敲除载体，并筛选出最佳敲除位点，获得了Apob基因敲除的亚克隆细胞，为进一步探索Apob基因在鸡肝中的功能奠定了基础。

课程思政的目标是在课程教学中实现知识传授、能力培养与价值引领的有机统一.以分子生物和基因工程课程为例,探讨在工程教育认证背景下开展课程思政教学的必要性,从制定课程思政教学目标、增强教师思政育人意识、深挖课程思政元素等方面开展课程思政教育的探索实践.同时,激发学生的科学精神、文化自信、家国情怀和使命担当,做到知识传授与价值引领、课程思政与专业认证、育人与育才的有机统一,实现铸魂育人的目标.

石蒜碱是一种普遍存在于石蒜科植物中的异喹啉类生物碱,现已发现其具有多种生物学活性.本文主要从抗癌、抗病毒、抗炎和调控脂质合成等方面阐述和探讨其作用机制,以期为石蒜碱在这些疾病中的潜在应用提供新的思路和依据.

Chemerin是由脂肪组织分泌的脂肪因子,可以调节脂肪细胞的代谢和免疫应答等生物学过程。本研究以鸡（Gallus gallus）肝脏组织总RNA为模板,采用逆转录PCR （reverse transcription PCR, RT-PCR）和c DNA末端快速扩增（rapid amplification of cDNA ends, RACE）方法获得鸡的Chemerin基因全长cDNA序列,应用qPCR检测鸡Chemerin基因mRNA在不同组织中的表达情况;构建Chemerin-Myc真核融合表达载体并转染LMH （Leghorn male hepatoma）细胞,用Western blot检测Chemerin-Myc融合蛋白表达情况;通过启动子片段截短实验,构建双荧光素酶报告基因载体,检测鸡Chemerin基因不同长度启动子片段的转录效率。结果表明,成功克隆鸡Chemerin基因,其cDNA全长为733 bp,编码162个氨基酸,其中5’UTR长度为55 bp,3’UTR长度为189 bp;Chemerin基因主要在肝脏、肾脏和脂肪组织中表达;成功构建真核融合表达载体pCMV-3Tag-9-gga-chemerin-Myc,转染鸡LMH细胞后,经Western blot检测发现,成功实现Chemerin和Myc的融合表达;构建的pGL3b-chemerin-F1/F2/F3/F4四个报告基因载体均显示荧光素酶活性比对照组极显著增加（P＜0.01）,F1和F3启动子中段的转录效率最高,pGL3b-chemerin-F1中的KLF5 （Kruppel-like factor 5）结合位点定点突变后,pGL3b-chemerin-mutF1的荧光素酶活性极其显著降低（P＜0.001）,在LMH细胞中干扰KLF5表达基因后,Chemerin基因的表达量极显著下降（P＜0.01）,初步证明在鸡Chemerin基因启动子区F1片段中存在正调控因子KLF5结合的顺式元件。本研究为进一步探讨鸡的Chemerin基因的功能提供了基础资料。

1. Duck meat is considered a delicacy, but choosing the best meat is problematic. Caveolin-3 (CAV-3) is a muscle-specific protein marker in animals. The goal of the current study was to detect the characteristics ofCAV-3gene in ducks. 2. Full-lengthCAV-3was acquired from ducks (Anas platyrhynchos)using reverse transcriptase PCR and rapid amplification of cDNA ends. DNAMAN software was used for homology comparisons. Quantitative reverse transcription-PCR, PCR-single-strand conformation polymorphism, and sequencing were used to determineCAV-3expression and polymorphism of a single nucleotide, respectively. The study examined four types of ducks, including Jinding, Chaohu, Cherry Valley, and Gaoyou ducks. 3. The study acquired 1066 bp ofCAV-3cDNA sequences, including a 456 bp complete open reading frame encoding 151 amino acids. Both coding sequences (CDSs) and translated amino acids exhibited highest homology withGallus gallus(CDS homology 91.67%, amino acids 94.04%), followed by mammalian species (CDS homology 79.0%, amino acids 78.0%). Single nucleotide polymorphism analysis revealed five mutations in exons (A489G, G501A, A557G, T563A, and A602G), and a C805T mutation in an intron. Among amplified polymorphic loci detected using primer 2, allele frequency was higher for A (489A501G507A563T602A) than B (489G501G507G563T602C) or C (489G501A507G563A602C). The highest occurred in Cherry Valley ducks (0.7587). Using primer 4, the M allele frequency was higher than that of the N allele.CAV-3was most highly expressed in the heart, followed by skeletal muscles. Additionally,CAV-3had higher expression in heart and breast muscle of overfed Muscovy ducks than control ducks, but no difference was seen in thigh muscle. 4.CAV-3in ducks had the highest homology with Gallus gallusCAV-3, and it could be used as a marker for muscle quality in ducks.

青钱柳(Cyclocarya paliurus)是胡桃科青钱柳属植物,近年来因其对改善糖脂代谢紊乱的良好作用日益成为研究热点.目前已从青钱柳中分离得到多种化合物,主要的生物活性成分包括多糖、三萜、黄酮、酚酸等.主要通过保护胰岛细胞、调节胰岛素信号通路、促进葡萄糖利用等发挥改善糖代谢的作用,并通过调控脂质代谢通路、抑制脂质过氧化等发挥改善脂代谢的作用.本文对青钱柳调节糖脂代谢活性成分及作用机制进行综述,为青钱柳防治糖尿病、高血脂、肥胖等代谢疾病提供参考.

固醇调控元件结合蛋白1(Sterol regulatory element-binding protein 1,SREBP1)通过调控脂肪生成相关的酶基因的转录,在机体脂类代谢调控中起着重要作用.本研究以21日龄的鸡胚肝组织的cDNA为模板,通过PCR分段克隆Srebp1全长基因的编码区片段,采用生物信息学分析其蛋白质功能结构域,将SREBP1功能结构域的编码片段克隆入原核表达载体pCold Ⅲ中,在大肠杆菌BL21(DE3)进行该功能片段的诱导表达.结果表明利用该基因内的Kpn Ⅰ和Not Ⅰ两个限制性酶切位点,成功将鸡Srebp1基因上分别长700,1 300,1 500 bp 3个片段依次插入pcDNA3.1(+)质粒的Hind Ⅲ和Xho Ⅰ酶切位点之间,获得全长Srebp1基因编码片段;在15℃和异丙基硫代半乳糖苷(Isopropylβ-D-Thiogalactoside,IPTG)的诱导条件下,pCold Ⅲ-gSrebp1-1125重组表达质粒在大肠杆菌BL21(DE3)中成功表达出重组蛋白gSREBP1-1125;SDS-PAGE电泳结果显示,重组蛋白gSREBP1-1125部分呈可溶性表达,但主要以包涵体存在.镍离子一步亲和层析法从菌体的裂解上清中获得了高纯度的重组蛋白;结合镍离子亲和层析,对重组蛋白进行固-液两相法复性,成功从包涵体中纯化得到高纯度的可溶性gSREBP1-1125功能多肽.鸡Srebp1真核表达栽体和功能片段重组蛋白的获得为进一步研究其结构、功能以及抗体的制备奠定了基础.

当今社会糖脂代谢紊乱的发生越发普遍,严重危害公共卫生健康.姜黄素(Curcumin)是一种从姜黄根茎中提取的天然活性成分,已广泛用于调节糖脂代谢紊乱研究.四氢姜黄素(Tetrahydrocurcumin,THC)是膳食姜黄素在体内的主要代谢产物,具有比姜黄素更好的抗氧化活性和胃肠道吸收,表现出多种优于姜黄素的生物学活性.因此,主要对四氢姜黄素的活性结构、体内代谢及调节糖脂代谢紊乱作用进行综述,以期为四氢姜黄素的进一步研发提供依据.