Mice derived from in vitro αMEM-cultured preimplantation embryos exhibit postprandial hyperglycemia and higher inflammatory gene expression in peripheral leukocytes

Abstract

We examined whether peripheral leukocytes of mice derived from in vitro-αMEM cultured embryos and exhibiting type 2 diabetes had higher expression of inflammatory-related genes associated with the development of atherosclerosis. Also, we examined the impact of a barley diet on inflammatory gene expression. Adult mice were produced by embryo transfer, after culturing two-cell embryos for 48 h in either α-MEM (minimal essential media) or KSOM (potassium simplex optimized medium) control media. Mice were fed either a barley or rice diet for 10 weeks. Postprandial blood glucose, and mRNA levels of several inflammatory genes, including Tnfa and Nox2, in blood leukocytes were significantly higher in MEM mice fed a rice diet compared with control mice. Barley intake reduced expression of S100a8 and Nox2. In summary, MEM mice exhibited postprandial hyperglycemia and peripheral leukocytes with higher expression of genes related to development of atherosclerosis, and barley intake reduced some gene expression.

Consumption of barley ameliorates the diabetic steatohepatitis and reduces the high transforming growth factor β expression in mice grown in α-minimum essential medium in vitro as embryos

“…In a previous study, we demonstrated that mice derived from embryos cultured in MEM exhibit T2DM and high postprandial inflammatory gene expression in peripheral leukocytes, compared to mice derived from embryos cultured in potassium simplex-optimized medium (KSOM) in vitro [ 6 ]. Furthermore, the expression of some inflammatory genes in peripheral leukocytes was reduced by feeding these mice a diet containing barley.…”

“…In the present study, we used same MEM and KSOM mice produced by the Kiwa Laboratory Animal Co., Ltd. (Wakayama, Japan) to analyze the development of steatohepatitis. The MEM mouse model and KSOM control mice have been described previously [ 5 , 6 ]. Briefly, two-cell stage embryos were collected from the oviducts of female Institute of Cancer Research (ICR) mice after mating with male ICR mice, and cultured in either MEM (135–15175, Wako Pure Chemical Industries, Ltd., Osaka, Japan) or KSOM (ARK Resource, Kumamoto, Japan) control medium for 48 h. The compositions of the MEM and KSOM are shown in Table S1 .…”

“…In a previous study, we demonstrated that mice derived from embryos cultured in α-minimal essential medium (MEM) at the two-cell stage and then transplanted back into the uterus (MEM mice) showed marked hyperglycemia and were slightly overweight [ 5 ]. MEM mice also exhibited higher expression of genes encoding proinflammatory molecules, including the integrins Cd11a, Cd11b, Cd11c and Cd18, interleukin (IL)-1 beta ( Il1b ) , S100a6 , and tumor necrosis factor alpha ( Tnfa ), in peripheral leukocytes [ 6 ]. It has also been reported that the serum concentration of myeloperoxidase-DNA, a marker of neutrophil extracellular traps that indicates the aggregation of inflammatory proteins in the blood, was higher in patients with NASH than in healthy individuals [ 7 ].…”

Table_1.xlsx

“…We have previously demonstrated that mice developed from embryos cultured in vitro in a-MEM (MEM mice) exhibit T2DM with postprandial hyperglycemia and non-alcoholic steatohepatitis, in contrast to mice developed from embryos cultured in vitro in potassium simplex optimized medium (KSOM) (23). Barley intake for 10 weeks ameliorated nonalcoholic steatohepatitis in MEM mice (21).…”

“…This animal study was approved by the Ethics Committee of the University of Yamanashi (approval number A30-24) and was performed according to the institutional animal experiment guidelines. The mice were decapitated, and samples were collected from one mouse at a time in the order of MR, MB, and KC to ensure that each group had similar mean dissection times (9:00 am-3:00 pm) (21,23). Kidney tissue samples were collected and weighed, and the right kidney tissues were snapfrozen in liquid nitrogen and stored at −80°C until use for qRT-PCR and western blotting.…”