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Alteration of gene expression patterns in the liver and spleen following exposure to nucleoprotein diet in the mouse LPS injection model

ABSTRACT: A diet rich in nucleic acids and protamin protein, termed as nucleoprotein (NP), has been attracting a great deal of attention in food science for its beneficial effects. In the present study, we performed a global gene expression profiling in mice fed with NP diet rich in nucleoproteins from the salmon testis. Since our recent research has revealed anti-inflammatory effect of the NP diet, we induced inflammation by lipopolysaccharide (LPS) injection in the mice for this analysis study. Mice were fed with NP diet for 4 weeks followed by a single injection of LPS. The liver and spleen were removed 6 h post-LPS injection. Total RNA extracted from livers (L) and spleens (S) was pooled in each group (control and NP diet), prior to DNA microarray analysis (Agilent mouse whole genome 4 x 44K). Results revealed 322 & 702 up (>1.5 fold)- and down (<0.75 fold)-regulated genes in the liver, and 325 & 501 up- and down-regulated genes in the spleen, respectively following NP diet. Analysis of genes related to inflammation suggests increased activity of immune function during acute period of LPS injection, which may contribute to early demise of inflammation. NP diet can be expected to be useful for inhibition of inflammatory reactions whose over-accumulation is thought to be related to the acceleration of aging process.

ORGANISM(S): Mus musculus

PROVIDER: GSE28176 | GEO | 2011/04/01

SECONDARY ACCESSION(S): PRJNA139525

REPOSITORIES: GEO

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Project description:A diet rich in nucleic acids and protamin protein, termed as nucleoprotein (NP), has been attracting a great deal of attention in food science for its beneficial effects. In the present study, we performed a global gene expression profiling in mice fed with NP diet rich in nucleoproteins from the salmon testis. Since our recent research has revealed anti-inflammatory effect of the NP diet, we induced inflammation by lipopolysaccharide (LPS) injection in the mice for this analysis study. Mice were fed with NP diet for 4 weeks followed by a single injection of LPS. The liver and spleen were removed 6 h post-LPS injection. Total RNA extracted from livers (L) and spleens (S) was pooled in each group (control and NP diet), prior to DNA microarray analysis (Agilent mouse whole genome 4 x 44K). Results revealed 322 & 702 up (>1.5 fold)- and down (<0.75 fold)-regulated genes in the liver, and 325 & 501 up- and down-regulated genes in the spleen, respectively following NP diet. Analysis of genes related to inflammation suggests increased activity of immune function during acute period of LPS injection, which may contribute to early demise of inflammation. NP diet can be expected to be useful for inhibition of inflammatory reactions whose over-accumulation is thought to be related to the acceleration of aging process. To investigate the effect of NP, we added S-nuclegen® at a concentration of 1.2% into CLEA basic purified diet (CLEA JAPAN, Inc., Tokyo, Japan) known to include nucleic acids (NAs) at much lower amounts than standard diet. By HPLC analysis CLEA basic purified diet (low NP), and 1.2% NP diet included 0.03%, and 0.5% NAs, respectively. Male C57BL/6J mice (7 weeks) were fed with NP diet for 4 weeks followed by a single injection of LPS at a dose of 10 mg/kg. The liver and spleen were removed 6 h post-LPS injection. Total RNA extracted from livers and spleens was pooled in each group (control and NP diet), prior to DNA microarray analysis (Agilent mouse whole genome 4 x 44K).

Project description:Background: Disease severity of autosomal dominant polycystic kidney disease (ADPKD) is influenced by diet. Dietary protein, a recognized cyst-accelerating factor, is catabolized into amino acids (AA) and delivered to the kidney leading to renal hypertrophy. Injury-induced hypertrophic signaling in ADPKD results in increased macrophage (MФ) activation and inflammation followed by cyst growth. We hypothesize that the cystogenesis-prompting effects of HP diet are caused by increased delivery of specific AA to the kidney, ultimately stimulating MФs to promote cyst progression. Methods: Pkd1 flox/flox mice with and without Cre (CAGG-ER) were given tamoxifen to induce global gene deletion (Pkd1KO). Pkd1KO mice were fed either a low (LP; 6%), normal (NP; 18%), or high (HP; 60%) diet for 1 week (early) or 6 weeks (chronic). Mice were then euthanized and tissues were used for histology, immunofluorescence and various biochemical assays. One week fed kidney tissue was cell sorted to isolate tubular epithelial cells for RNA sequencing. Results: Chronic dietary protein load in Pkd1KO mice increased kidney weight, number of kidney infiltrating and resident MФs, chemokines, cytokines and cystic index compared to LP diet fed mice. Accelerated cyst growth induced by chronic HP were attenuated by liposomal clodronate-mediated MФ depletion. Early HP diet fed Pkd1KO mice had larger cystic kidneys compared to NP or LP fed counterparts, but without increases in the number of kidney MФs, cytokines, or markers of tubular injury. RNA sequencing of tubular epithelial cells in HP compared to NP or LP diet group revealed increased expression of sodium-glutamine transporter Snat3, chloride channel Clcnka, and gluconeogenesis marker Pepck1, accompanied by increased excretion of urinary ammonia, a byproduct of glutamine. Conclusion: Chronic dietary protein load-induced renal hypertrophy and accelerated cyst growth in Pkd1KO mice is dependent on both infiltrating and resident MФ recruitment and subsequent inflammatory response. Early cyst expansion by HP diet, however, is relient on increased delivery of glutamine to kidney epithelial cells, driving downstream metabolic changes prior to inflammatory provocation.

Project description:The grass-fed cattle obtain nutrients directly from pastures containing limited assimilable energy but abundant amount of fiber; by contrast, grain-fed steers receive a diet that is comprised mainly of grains and serves as an efficient source of high-digestible energy. Besides energy, these two types of diet differ in a large number of nutritional components. Additionally, animals maintained on rich-energy regimen are more likely to develop metabolic disorders and infectious diseases than pasture raised individuals. Thus, we hypothesize that spleenâ??the main immune organâ??may function differently under disparate regimes. The objective of this study was to find the differentially expressed genes in the spleen of grass-fed and grain-fed steers, and furtherly explore the potential involved biopathways. Through RNA sequencing (RNA-Seq), we detected 123 differentially expressed genes. Based on these genes, we performed an Ingenuity Pathway Analysis (IPA) and identified 9 significant molecular networks and 13 enriched biological pathways. Two of the pathways, Nur77 signaling in T lymphocytes and calcium-induced T lymphocyte apoptosis which are immune related, contain a pair of genes HLA-DRA and NR4A1 with dramatically altered expression level. Collectively, our results provided valuable insights into understanding the molecular mechanism of spleen under varied feeding regimens. We collected spleen samples from two randomly chosen animals per group, totaling four samples. The animals were born and raised at the Wye Angus farm, which has produced genetically similar progenies. The genetic resemblance among individuals permitted us to better control the variation between experimental individuals, constituting an excellent resource to perform scientific research. All animals included in this study received the same diet until weaning. Next, we assigned the animals to one certain diet at random, and exclusively raised them under that regimen until termination. The diet of grain-fed group consisted of soybean, shelled corn, corn silage and trace minerals. The grass-fed steers normally received alfalfa harvested from land without any fertilizers, pesticides or other chemicals; during wintertime, bailage was supplied. Grass-fed individuals ate no animal, agricultural or industrial byproducts and never consumed any type of grain. Grain-fed animals reached the market weight around 14 month-old; however, grass-fed steers needed approximately 200 additional days to achieve the same weight. Immediately after termination at the Old Line Custom Meat Company (Baltimore, MD), a small piece of spleen was incised, washed and frozen at -80Â°C for posterior processing.

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Alteration of gene expression patterns in the liver and spleen following exposure to nucleoprotein diet in the mouse LPS injection model

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