Identification of stably expressed Internal Control Genes (ICGs) for normalization of expression data in liver of C57BL/6 mice injected with beta casomorphins.

In recent years, beta-casomorphin peptides (BCM7/BCM9) derived from the digestion of cow milk have drawn a lot of attention world over because of their proposed impact on human health. In order to evaluate the transcriptional modulation of target genes through RT-qPCR in response to these peptides, availability of appropriate reference or internal control genes (ICGs) will be the key. The present study was planned to identify a panel of stable ICGs in the liver tissue of C57BL/6 mice injected with BCM7/BCM9 cow milk peptides for 3 weeks. A total of ten candidate genes were evaluated as potential ICGs by assessing their expression stability using software suites; geNorm, NormFinder and BestKeeper. The suitability of the identified ICGs was validated by assessing the relative expression levels of target genes, HP and Cu/Zn SOD. Based on geNorm, PPIA and SDHA gene pair was identified to be most stably expressed in liver tissue during the animal trials. Similarly, NormFinder analysis also identified PPIA as the most stable gene. BestKeeper analysis showed crossing point SD value for all the genes in the acceptable range that is closer to 1. Overall, the study identified a panel of stable ICGs for reliable normalization of target genes expression data in mice liver tissues during BCM7/9 peptides trial.

Co-Administration of Soluble Fibres and Lactobacillus casei NCDC19 Fermented Milk Prevents Adiposity and Insulin Resistance Via Modulation of Lipid Mobilization Genes in Diet-Induced Obese Mice.

BACKGROUND: Numerous reports explaining the beneficial health effects of soluble fibres and probiotics on lifestyle disorders have been published. However, a little information is available on coadministration of soluble fibres such as gum acacia & inulin and probiotic lactobacilli. Therefore, in the present study, we have evaluated the synergistic effects of soluble fibres and probiotic fermented milk on adiposity, insulin resistance and dyslipidemia in C57BL/6 mice fed high-fat and sucrose diet for 18 weeks. OBJECTIVE: To explore the synergistic effect of soluble fibres (gum acacia/inulin) and Lactobacillus casei NCDC19 fermented milk on adiposity, insulin resistance and lipid mobilization genes in dietinduced obese mice. METHODS: C57BL/6 mice were divided randomly into three groups (n = 9/group) according to their body weights. The HFS group was fed high-fat and sucrose diet, the HFS-GFM group was fed HFS diet incorporated with gum acacia (7%, w/w) along with L. casei NCDC19 fermented milk and HFSIFM group was fed HFS diet incorporated with inulin (7%, w/w) along with L. casei NCDC19 fermented milk. RESULTS: At the end of the experiment, final body weight, epididymal fat (E.fat) weight, and adipocyte size were found to be lower in groups received either gum acacia or inulin in combination with L. casei NCDC19 fermented milk (HFS-GFM or HFS-IFM). Also, fasting blood glucose, serum insulin, triglycerides, and VLDL-cholesterol levels were decreased significantly in both HFS-GFM and HFSIFM fed groups. Furthermore, relative mRNA expression of genes (cpt1, foxa2, pgc1ß, and pparα) related to fatty acid oxidation enhanced significantly in the liver. In E.fat pad, expression of adiponectin was upregulated, whereas, leptin expression was reduced considerably. Also, expression of fasting-induced adipose factor enhanced significantly in the distal ileum of mice in HFS-GFM and HFS-IFM groups. CONCLUSION: Overall, we demonstrate that co-administration of soluble fibres viz. gum acacia, inulin and L. casei NCDC19 fermented milk exhibited the anti-adiposity effects, improved insulin sensitivity and dyslipidemia in mice via modulation of lipid mobilization genes.

Dietary incorporation of whey protein isolate and galactooligosaccharides exhibits improvement in glucose homeostasis and insulin resistance in high fat diet fed mice.

BACKGROUND: This study was planned to investigate the effectiveness of the whey protein isolate (WPI) of high purity and a galactooligosaccharides (GOS) preparation on glucose homeostasis and insulin resistance in high fat diet (HFD) (45.47% energy from fat) fed conditions in C57BL/6J mice. METHODS: Fasting blood glucose level, serum insulin, and glucagon-like peptide-1 (enzyme-linked immunosorbent assay) were measured; also, homeostasis model assessment of insulin resistance (HOMA-IR) was determined in different treatment groups. mRNA expression of gluconeogenesis genes in liver and small intestine tissues was analyzed by quantitative real time-polymerase chain reaction. RESULTS: Dietary incorporation of WPI and GOS was observed to significantly resist (P < 0.001) the HFD-induced increase in blood glucose levels indicating a mitigating effect on glycemic load. It is important to note that no additive effects of administration of WPI and GOS could be observed. The administration of WPI and GOS exhibited maximum resistance (37.8%) to the rise in insulin level. Thus, the resistance to the increase in HOMA-IR was also noticed on the dietary incorporation of two functional ingredients . The positive effects on mRNA expression of phosphoenolpyruvate carboxykinase and glucose 6-phosphatase could be detected in liver only. CONCLUSION: Both types of functional components exhibit potential to improve glucose homeostasis under HFD fed conditions. Resistance to HFD-induced hyperinsulinemia and HOMA-IR is also recorded .