N-glycoproteomic analysis of Ginsenoside Rb1 on a hyperlipidemia rat model

Abstract Background: Ginsenoside Rb1, known as Renshen in traditional Chinese medicine, is one of the major bioactive saponins isolated from Panax ginseng C.A.Mey. N-glycosylation is the most common type of post-translational modification in cells. The widespread localization of N-glycosylated proteins (N-glycoproteins) between extracellular spaces and on the cell surfaces give them unique advantages as disease biomarkers and drug targets. Previous study found that Ginsenoside Rb1 could potentially play a preventive role in hyperlipidemia. This study aims to reveal the hypolipidemic effect at the protein modification level. Methods: 24 male SD rats were randomly devided into 3 groups: control group (CON), high fat diet group (HFD) and Ginsenoside Rb1 group (Rb1). Both HFD and Rb1 groups were fed with high-fat diet for 12 weeks. The Rb1 group started intragastric administering Ginsenoside Rb1 200 mg·kg -1 ·d -1 at 5th week for 8 weeks, while the CON and HFD group the same amount of normal saline for the same amount of time. Lipid levels and liver histology were assayed to evaluate the effects of Ginsenoside Rb1 intake on hyperlipidemia rats. Furthermore, the workflow by combination of isotope TMT labeling, HILIC enrichment, and high-resolution LC-MS/MS analysis were employed to exploring the mechanisms of regulation role in hyperlipidemia rats.Results: The histopathologic characteristics and biochemical data shows that Ginsenoside Rb1 exhibited regulative effects on hyperlipidemia rats. After being analyzed by N-glycoproteomic, 98 differential N-glycosylation sites on 53 glycoproteins between 2 comparison groups (HFD: CON, Rb1: HFD) were identified. Analyses of N-glycosylation sites distribution found that albumin (Alb) and Serpinc1 were most heavily modified with 6 N-glycosylation sites changed in this work. GO enrichment analysis showed that differential modified glycoproteins were involved in inflammatory response, cellular iron ion homeostasis and positive regulation of cholesterol efflux etc. biosynthetic process. Complement and coagulation cascades was the most significant enriched in the KEGG pathway enrichment analysis. Conclusions: This study presents a comprehensive analysis of a new set of N-glycoproteins which are altered by Ginsenoside Rb1 and offers some valuable clues for novel mechanistic insights into the ragulative mechanism of Ginsenoside Rb1. Results from N-glycoproteomic suggest that to suppress hyperlipidemia, Rb1 may regulates N-glycosylation of Alb, Serpinc1, PON1, Lrp1, Cp and THBS1, as well as differentially modified glycoproteins in complement and coagulation cascades, which in turn improve the imbalance of lipid homeostasis.

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CCDC114, DNAI2 and TOP2A involves in the effects of tibolone treatment on postmenopausal endometrium

BMC Women s Health ◽

10.1186/s12905-020-01156-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yanhua Lv ◽

Yanqing Liu ◽

Yueqiang Wang ◽

Fanrong Kong ◽

Qiuxiang Pang ◽

...

Keyword(s):

Postmenopausal Women ◽

Molecular Mechanisms ◽

Ion Homeostasis ◽

Enrichment Analysis ◽

Control Group ◽

Pathway Enrichment Analysis ◽

Ppi Network ◽

Target Interaction ◽

Protein Protein Interaction ◽

Movement Function

Abstract Background This study aimed to explore the molecular mechanisms of tibolone treatment in postmenopausal women. Methods The gene set enrichment profile, GSE12446, which includes 9 human endometrial samples from postmenopausal women treated with tibolone (tibolone group) and 9 control samples (control group), was downloaded from GEO database for analysis. Differentially expressed genes (DEGs) in tibolone vs. control groups were identified and then used for function and pathway enrichment analysis. Protein–protein interaction (PPI) network and module analyses were also performed. Finally, drug–target interaction was predicted for genes in modules, and then were validated in Pubmed. Results A total of 238 up-regulated DEGs and 72 down-regulated DEGs were identified. These DEGs were mainly enriched in various biological processed and pathways, such as cilium movement (e.g., CCDC114 and DNAI2), calcium ion homeostasis, regulation of hormone levels and complement/coagulation cascades. PPI network contained 368 interactions and 166 genes, of which IGF1, DNALI1, CCDC114, TOP2A, DNAH5 and DNAI2 were the hue genes. A total of 96 drug–gene interactions were obtained, including 94 drugs and eight genes. TOP2A and HTR2B were found to be targets of 28 drugs and 38 drugs, respectively. Among the 94 obtained drugs, only 12 drugs were reported in studies, of which 7 drugs (e.g., epirubicin) were found to target TOP2A. Conclusions CCDC114 and DNAI2 might play important roles in tibolone-treated postmenopausal women via cilium movement function. TOP2A might be a crucial target of tibolone in endometrium of postmenopausal women.

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Genome-wide identification and characterization of perirenal adipose tissue microRNAs in rabbits fed a high-fat diet

Bioscience Reports ◽

10.1042/bsr20204297 ◽

2021 ◽

Author(s):

Jie Wang ◽

Jiahao Shao ◽

Yanhong Li ◽

Mauricio A Elzo ◽

Xianbo Jia ◽

...

Keyword(s):

Intermediate Filament ◽

High Fat Diet ◽

Metabolic Regulation ◽

Target Genes ◽

Enrichment Analysis ◽

Normal Diet ◽

Pathway Enrichment Analysis ◽

High Fat ◽

Cytoskeleton Organization ◽

Kegg Pathways

MicroRNAs (miRNAs) are a class of endogenous single-stranded RNA molecules that play an important role in gene regulation in animals by pairing with target gene mRNA. Extensive evidence shows that miRNAs are key players in metabolic regulation and the development of obesity. However, the systemic understanding of miRNAs in the adipogenesis of obese rabbit need further investigate. Here, seven small RNA libraries from rabbits fed either a standard normal diet (SND; n = 3) or high-fat diet (HFD; n = 4) were constructed and sequenced. Differentially expressed (DE) miRNAs were identified using the edgeR data analysis package from R. Software miRanda and RNAhybrid were used to predict the target genes of miRNAs. To further explore the functions of DE miRNAs, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. A total of 81,449,996 clean reads were obtained from the seven libraries, of which, 52 known DE miRNAs (24 up-regulated, 28 down-regulated) and 31 novel DE miRNAs (14 up-regulated, 17 down-regulated) were identified. GO enrichment analysis revealed that the DE miRNAs target genes were involved in intermediate filament cytoskeleton organization, intermediate filament-based process, and alpha-tubulin binding. DE miRNAs were involved in p53 signaling, linoleic acid metabolism, and other adipogenesis-related KEGG pathways. Our study further elucidates the possible functions of DE miRNAs in rabbit adipogenesis, contributing to the understanding of rabbit obesity.

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Cardiac Transcriptome Analysis Reveals Nr4a1 Mediated Glucose Metabolism Dysregulation in Response to High-Fat Diet

Genes ◽

10.3390/genes11070720 ◽

2020 ◽

Vol 11 (7) ◽

pp. 720

Author(s):

Lihui Men ◽

Wenting Hui ◽

Xin Guan ◽

Tongtong Song ◽

Xuan Wang ◽

...

Keyword(s):

Glucose Metabolism ◽

Transcriptome Analysis ◽

High Fat Diet ◽

Critical Role ◽

Enrichment Analysis ◽

Pathway Enrichment Analysis ◽

High Fat ◽

Altered Expression ◽

Increased Risk ◽

H9c2 Cardiomyocytes

Obesity is associated with an increased risk of developing cardiovascular disease (CVD), with limited alterations in cardiac genomic characteristics known. Cardiac transcriptome analysis was conducted to profile gene signatures in high-fat diet (HFD)-induced obese mice. A total of 184 differentially expressed genes (DEGs) were identified between groups. Based on the gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs, the critical role of closely interlocked glucose metabolism was determined in HFD-induced cardiac remodeling DEGs, including Nr4a1, Fgf21, Slc2a3, Pck1, Gck, Hmgcs2, and Bpgm. Subsequently, the expression levels of these DEGs were evaluated in both the myocardium and palmitic acid (PA)-stimulated H9c2 cardiomyocytes using qPCR. Nr4a1 was highlighted according to its overexpression resulting from the HFD. Additionally, inhibition of Nr4a1 by siRNA reversed the PA-induced altered expression of glucose metabolism-related DEGs and hexokinase 2 (HK2), the rate-limiting enzyme in glycolysis, thus indicating that Nr4a1 could modulate glucose metabolism homeostasis by regulating the expression of key enzymes in glycolysis, which may subsequently influence cardiac function in obesity. Overall, we provide a comprehensive understanding of the myocardium transcript molecular framework influenced by HFD and propose Nr4a1 as a key glucose metabolism target in obesity-induced CVD.

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Unveil the transcriptome alteration of POMC neuron in diet-induced obesity

10.21203/rs.2.19329/v1 ◽

2019 ◽

Author(s):

Peng Lyu ◽

Zhishun Huang ◽

Qingjun Feng ◽

Yongfu Su ◽

Mengying Zheng ◽

...

Keyword(s):

Cell Cycle ◽

High Fat Diet ◽

Arcuate Nucleus ◽

Enrichment Analysis ◽

Sphingolipid Metabolism ◽

Pathway Enrichment Analysis ◽

High Fat ◽

Transcriptomic Profiling ◽

Diet Induced Obesity ◽

Underlying Mechanisms

Abstract Background: Loss of neuron homeostasis in the Arcuate nucleus (ARC) is suggested to be responsible for the development diet-induced-obesity (DIO). We previously reported that loss of Rb1 gene compromised the homeostasis of anorexigenic POMC neurons in ARC and induced obesity in mice.Method: To shed light on how DIO develops, we propose to analyze the transcriptomic alteration of POMC neurons in mice following high fat die (HFD) feeding. We isolated the POMC neurons from established DIO mice and performed transcriptomic profiling on them by RNA-seq.Results: A total of 1,066 genes (628 up-regulated and 438 down-regulated) were identified as differentially expressed genes (DEGs). Pathway enrichment analysis with these DEGs further revealed that ‘cell cycle’, ‘apoptosis’, ‘chemokine signalling’ and ‘sphingolipid metabolism’ pathways were correlated with the development of DIO. Moreover, we validated that the pRb protein, key regulator of ‘cell cycle pathway’, was inactivated by phosphorylation in POMC neurons with HFD feeding. Importantly, reversal of deregulated cell cycle by stereotaxic delivering of the unphosphorylated pRb∆P in ARC significantly meliorated the DIO. Together, our study provides insights into the mechanisms related to the loss of homeostasis of POMC neurons in DIO, and suggests pRb phosphorylation as a potential intervention target to treat DIO. Conclusion: The Arcuate nucleus is the material basis that controlled energy balance and glucose metabolism, which is vulnerable to high-fat-diet (HFD) in diet-induced-obesity (DIO). In this study, we conducted transcriptomic profiling in anorexigenic POMC neurons of ARC with HFD to disclose the underlying mechanisms related with the homeostasis maintenance and the development of DIO. Importantly, we suggest that DIO could be prevented of treated by reversal of the deregulated cell cycle in POMC neurons through targeting pRb phosphorylation. Keywords: High-Fat-Diet (HFD); Diet-Induced Obesity (DIO); POMC neuron; Neuron homeostasis; pRb phosphorylation

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Genome-Wide DNA Methylation Changes of Perirenal Adipose Tissue in Rabbits Fed a High-Fat Diet

Animals ◽

10.3390/ani10122213 ◽

2020 ◽

Vol 10 (12) ◽

pp. 2213

Author(s):

Jiahao Shao ◽

Xue Bai ◽

Ting Pan ◽

Yanhong Li ◽

Xianbo Jia ◽

...

Keyword(s):

Dna Methylation ◽

Adipose Tissue ◽

High Fat Diet ◽

Enrichment Analysis ◽

Epigenetic Mechanism ◽

Pathway Enrichment Analysis ◽

High Fat ◽

Perirenal Adipose Tissue ◽

Pathway Enrichment ◽

Genome Wide

DNA methylation is an epigenetic mechanism that plays an important role in gene regulation without an altered DNA sequence. Previous studies have demonstrated that diet affects obesity by partially mediating DNA methylation. Our study investigated the genome-wide DNA methylation of perirenal adipose tissue in rabbits to identify the epigenetic changes of high-fat diet-mediated obesity. Two libraries were constructed pooling DNA of rabbits fed a standard normal diet (SND) and DNA of rabbits fed a high-fat diet (HFD). Differentially methylated regions (DMRs) were identified using the option of the sliding window method, and online software DAVID Bioinformatics Resources 6.7 was used to perform Gene Ontology (GO) terms and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis of DMRs-associated genes. A total of 12,230 DMRs were obtained, of which 2305 (1207 up-regulated, 1098 down-regulated) and 601 (368 up-regulated, 233 down-regulated) of identified DMRs were observed in the gene body and promoter regions, respectively. GO analysis revealed that the DMRs-associated genes were involved in developmental process (GO:0032502), cell differentiation (GO:0030154), and lipid binding (GO:0008289), and KEGG pathway enrichment analysis revealed the DMRs-associated genes were enriched in linoleic acid metabolism (KO00591), DNA replication (KO03030), and MAPK signaling pathway (KO04010). Our study further elucidates the possible functions of DMRs-associated genes in rabbit adipogenesis, contributing to the understanding of HFD-mediated obesity.

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Genome-wide Identification and Characterization of Perirenal Adipose Tissue MiRNAs in Rabbits Fed With a High-fat Diet

10.21203/rs.3.rs-44842/v1 ◽

2020 ◽

Author(s):

Jie Wang ◽

Jiahao Shao ◽

Yanhong Li ◽

Mauricio A. Elzo ◽

Xianbo Jia ◽

...

Keyword(s):

Intermediate Filament ◽

High Fat Diet ◽

Target Genes ◽

Enrichment Analysis ◽

Normal Diet ◽

Pathway Enrichment Analysis ◽

High Fat ◽

Alpha Tubulin ◽

Cytoskeleton Organization ◽

Kegg Pathways

Abstract Background MicroRNAs (miRNAs) are a class of endogenous single-stranded RNA molecules that play an important role in gene regulation in animals by pairing with target gene mRNAs. However, the functions of miRNAs in the adipogenesis of obese rabbits are poorly understood. Methods Six small RNA libraries from rabbits under a standard normal diet (SND; n = 3) and a high-fat diet (HFD; n = 3) were constructed and sequenced. Differentially expressed (DE) miRNAs were identified using the edgeR data analysis package from R. Software miRanda and RNAhybrid were used to predict the target genes of miRNAs. To further explore the functions of DE miRNAs, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Results A total of 69577441 clean reads were obtained from the six libraries, of which, 52 known DE miRNAs (24 up-regulated, 28 down-regulated) and 31 novel DE miRNAs (14 up-regulated, 17 down-regulated) were identified. GO enrichment analysis revealed that the DE miRNAs target genes were involved in intermediate filament cytoskeleton organization, intermediate filament-based process, and alpha-tubulin binding. DE miRNAs were involved in p53 signaling, linoleic acid metabolism, and other adipogenesis-related KEGG pathways. Conclusions Our study further elucidates the possible functions of DE miRNAs in rabbit adipogenesis, contributing to the understanding of rabbit obesity.

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The role of purslane in modulating diverse effects of high fat diet on biochemical, histological, and molecular parameters of rats’ liver

Brazilian Journal of Biology ◽

10.1590/1519-6984.248755 ◽

2023 ◽

Vol 83 ◽

Author(s):

A. M. Mousa ◽

M. E. Taha ◽

SH. M. ELdeighdye ◽

A. M. Kamal

Keyword(s):

High Fat Diet ◽

Potential Role ◽

Density Lipoprotein ◽

Diet Group ◽

Control Group ◽

Hypolipidemic Effect ◽

High Fat ◽

Hypolipidemic Agent ◽

Harmful Effects

Abstract Consuming a high-fat diet causes a harmful accumulation of fat in the liver, which may not reverse even after switching to a healthier diet. Different reports dealt with the role of purslane as an extract against high-fat diet; meanwhile, it was necessary to study the potential role of fresh purslane as a hypolipidemic agent. This study is supposed to investigate further the potential mechanism in the hypolipidemic effect of fresh purslane, by measuring cholesterol 7a-hydroxylase (CYP7A1) and low-density lipoprotein receptor (Ldlr). Rats were divided into two main groups: the first one is the normal control group (n=7 rats) and the second group (n=28 rats) received a high fat diet for 28 weeks to induce obesity. Then the high fat diet group was divided into equal four subgroups. As, the positive control group still fed on a high fat diet only. Meanwhile, the other three groups were received high-fat diet supplemented with a different percent of fresh purslane (25, 50 and 75%) respectively. At the end of the experiment, rats were sacrificed and samples were collected for molecular, biochemical, and histological studies. Current study reported that, supplementation of fresh purslane especially at a concentration of 75% play an important role against harmful effects of high-fat diet at both cellular and organ level, by increasing CYP7A1 as well as Ldlr mRNA expression. Also, there were an improvement on the tested liver functions, thyroid hormones, and lipid profile. Fresh purslane plays the potential role as a hypolipidemic agent via modulation of both Ldlr and Cyp7A, which will point to use fresh purslane against harmful effects of obesity.

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Anti-Obesity and Hypocholesterolemic Actions of Protamine-Derived Peptide RPR (Arg-Pro-Arg) and Protamine in High-Fat Diet-Induced C57BL/6J Mice

Nutrients ◽

10.3390/nu13082501 ◽

2021 ◽

Vol 13 (8) ◽

pp. 2501

Author(s):

Maihemuti Mijiti ◽

Ryosuke Mori ◽

Bingyu Huang ◽

Kenichiro Tsukamoto ◽

Keisuke Kiriyama ◽

...

Keyword(s):

Adipose Tissue ◽

High Fat Diet ◽

Fecal Excretion ◽

Control Group ◽

High Fat ◽

Tissue Weight ◽

Cholesterol Levels ◽

Adipose Tissue Weight ◽

Fas Mrna ◽

Hypocholesterolemic Peptide

Dietary protamine can ameliorate hyperlipidemia; however, the protamine-derived active peptide and its hypolipidemic mechanism of action are unclear. Here, we report the discovery of a novel anti-obesity and hypocholesterolemic peptide, RPR (Arg-Pro-Arg), derived from protamine in mice fed a high-fat diet for 50 days. Serum cholesterol levels were significantly lower in the protamine and RPR groups than in the control group. White adipose tissue weight was significantly decreased in the protamine and RPR groups. The fecal excretion of cholesterol and bile acid was significantly higher in the protamine and RPR groups than in the control group. We also observed a significant decrease in the expression of hepatic SCD1, SREBP1, and adipocyte FAS mRNA, and significantly increased expression of hepatic PPARα and adipocyte PPARγ1 mRNA in the protamine group. These findings demonstrate that the anti-obesity effects of protamine are linked to the upregulation of adipocyte PPARγ1 and hepatic PPARα and the downregulation of hepatic SCD1 via SREBP1 and adipocyte FAS. RPR derived from protamine has a crucial role in the anti-obesity action of protamine by evaluating the effective dose of adipose tissue weight loss.

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Aberrant expression of HDL-bound microRNA induced by a high-fat diet in a pig model: implications in the pathogenesis of dyslipidaemia

BMC Cardiovascular Disorders ◽

10.1186/s12872-021-02084-5 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Guoyuan Sui ◽

Lianqun Jia ◽

Nan Song ◽

Dongyu Min ◽

Si Chen ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

High Fat Diet ◽

Enrichment Analysis ◽

Diet Group ◽

High Fat ◽

Positive Regulation ◽

Aberrant Expression ◽

Balanced Diet ◽

Mirna Sequencing

Abstract Background A high-fat diet can affect lipid metabolism and trigger cardiovascular diseases. A growing body of studies has revealed the HDL-bound miRNA profiles in familial hypercholesterolaemia; in sharp contrast, relevant studies on high-fat diet-induced dyslipidaemia are lacking. In the current study, HDL-bound miRNAs altered by a high-fat diet were explored to offer some clues for elucidating their effects on the pathogenesis of dyslipidaemia. Methods Six pigs were randomly divided into two groups of three pigs each, namely, the high-fat diet and the balanced diet groups, which were fed a high-fat diet and balanced diet separately for six months. HDL was separated from plasma, which was followed by dissociation of the miRNA bound to HDL. miRNA sequencing of the isolated miRNA was performed to identify the differential expression profiles between the two groups, which was validated by real-time PCR. TargetScan, miRDB, and miRWalk were used for the prediction of genes targeted by the differential miRNAs. Results Compared with the balanced diet group, the high-fat diet group had significantly higher levels of TG, TC, LDL-C and HDL-C at six months. miRNA sequencing revealed 6 upregulated and 14 downregulated HDL-bound miRNAs in the high-fat diet group compared to the balanced diet group, which was validated by real-time PCR. GO enrichment analysis showed that dysregulated miRNAs in the high-fat diet group were associated with the positive regulation of lipid metabolic processes, positive regulation of lipid biosynthetic processes, and positive regulation of Ras protein signal transduction. Insulin resistance and the Ras signalling pathway were enriched in the KEGG pathway enrichment analysis. Conclusions Twenty HDL-bound miRNAs are significantly dysregulated in high-fat diet-induced dyslipidaemia. This study presents an analysis of a new set of HDL-bound miRNAs that are altered by a high-fat diet and offers some valuable clues for novel mechanistic insights into high-fat diet-induced dyslipidaemia. Further functional verification study using a larger sample size will be required.

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Maternal tadalafil therapy for fetal growth restriction prevents non-alcoholic fatty liver disease and adipocyte hypertrophy in the offspring

Scientific Reports ◽

10.1038/s41598-020-80643-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Takuya Kawamura ◽

Hiroaki Tanaka ◽

Ryota Tachibana ◽

Kento Yoshikawa ◽

Shintaro Maki ◽

...

Keyword(s):

Liver Disease ◽

Fatty Liver ◽

Fetal Growth ◽

Fetal Programming ◽

High Fat Diet ◽

Fatty Liver Disease ◽

Control Group ◽

High Fat ◽

Alcoholic Fatty Liver ◽

Alcoholic Fatty Liver Disease

AbstractWe aimed to investigate the effects of maternal tadalafil therapy on fetal programming of metabolic function in a mouse model of fetal growth restriction (FGR). Pregnant C57BL6 mice were divided into the control, L-NG-nitroarginine methyl ester (L-NAME), and tadalafil + L-NAME groups. Six weeks after birth, the male pups in each group were given a high-fat diet. A glucose tolerance test (GTT) was performed at 15 weeks and the pups were euthanized at 20 weeks. We then assessed the histological changes in the liver and adipose tissue, and the adipocytokine production. We found that the non-alcoholic fatty liver disease activity score was higher in the L-NAME group than in the control group (p < 0.05). Although the M1 macrophage numbers were significantly higher in the L-NAME/high-fat diet group (p < 0.001), maternal tadalafil administration prevented this change. Moreover, the epididymal adipocyte size was significantly larger in the L-NAME group than in the control group. This was also improved by maternal tadalafil administration (p < 0.05). Further, we found that resistin levels were significantly lower in the L-NAME group compared to the control group (p < 0.05). The combination of exposure to maternal L-NAME and a high-fat diet induced glucose impairment and non-alcoholic fatty liver disease. However, maternal tadalafil administration prevented these complications. Thus, deleterious fetal programming caused by FGR might be modified by in utero intervention with tadalafil.

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