Changes of Blood-Brain Barrier and Brain Parenchymal Protein Expression Levels of Mice under Different Insulin-Resistance Conditions Induced by High-Fat Diet

Ogata, Seiryo; Ito, Shingo; Masuda, Takeshi; Ohtsuki, Sumio

doi:10.1007/s11095-019-2674-8

Cited by 39 publications

(33 citation statements)

References 39 publications

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“…Western blot analysis of isolated brain capillary fraction was performed as described previously. 5 Briefly, each sample was separated on an SDS-polyacrylamide gel and blotted onto a PVDF membrane. We used following primary antibodies (Claudin-5, 35-2500, Thermo; Glut1, ab652, abcam; Mdr1, 13978, CST; β-actin, 8H10D10, CST) and HRP conjugated secondary antibodies (Goat anti rabbit IgG HRP, ab6721, abcam; Goat anti mouse IgG HRP, ab6789, abcam).…”

Section: Methodsmentioning

confidence: 99%

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Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis

Ogata

Ito

Masuda

et al. 2020

J Cereb Blood Flow Metab

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Isolated brain capillaries are essential for analyzing the changes of protein expressions at the blood–brain barrier (BBB) under pathological conditions. The standard brain capillary isolation methods require the use of at least five mouse brains in order to obtain a sufficient amount and purity of brain capillaries. The purpose of this study was to establish a brain capillary isolation method from a single mouse brain for protein expression analysis. We successfully isolated brain capillaries from a single frozen mouse brain by using a bead homogenizer in the brain homogenization step and combination of cell strainers and glass beads in the purification step. Western blot and proteomic analysis showed that proteins expressed at the BBB in mouse brain capillaries isolated by the developed method were more enriched than those isolated from a pool of five mouse brains by the standard method. By using the developed method, we further verified the changes in expression of BBB proteins in Glut1-deficient mouse. The developed method is useful for the analysis of various mice models with low numbers and enables us to understand, in more detail, the physiology and pathology of BBB.

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Section: Methodsmentioning

confidence: 99%

“…We previously reported that proteomic analysis is a useful technique for revealing protein expression levels in normal physiological conditions as well as under pathological conditions at the BBB. 5 To analyze the BBB, brain capillary isolation is an essential method because the volume of brain capillaries is only 0.1% of whole brain. 6…”

Section: Introductionmentioning

confidence: 99%

Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis

Ogata

Ito

Masuda

et al. 2020

J Cereb Blood Flow Metab

Self Cite

View full text Add to dashboard Cite

show abstract

“…The volume of hippocampus in patients with mild CI was smaller than that in healthy elderly people (De Leon et al, 2004;Hampel et al, 2008). Furthermore, in women aged 50-65 undergoing hormone therapy, insulin resistance (HOMA-IR) had a significant negative relationship with right and total hippocampal volume (Rasgon et al, 2011). A recent study assessed 120 adults without dementia and with long duration type 2 diabetes, the results indicated that asymmetric atrophy, with greater atrophy occurring in the right hippocampus, contributed to CI in type 2 diabetes (Milne et al, 2017).…”

Section: Discussionmentioning

confidence: 86%

“…Is there a direct correlation between capillaries in the hippocampus and the impairment of cognitive function? A recent study found that the expression levels of P-glycoprotein function at the bloodbrain barrier were reduced in high-fat diet mice at 2 weeks (Ogata, Ito, Masuda, & Ohtsuki, 2019). Using detailed cognitive assessments and 3-Tesla MRI using intravoxel incoherent motion (IVIM) MRI, Van Bussel Note: Data are presented as the mean ± SD and were analyzed using two-tailed t tests in independent samples; There were significant difference in the volumes after IHC stain and PAS stain.…”

Section: Discussionmentioning

confidence: 99%

Changes in hippocampal capillaries in transgenic type 2 diabetic mice: A stereological investigation

Zhao

Liu

et al. 2020

The Anatomical Record

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The cognitive dysfunction associated with type 2 diabetes mellitus (T2DM) has been widely studied, and many structures in the hippocampus, such as neurons and synapses, have been shown to play a crucial role in the cognitive decline. However, the mechanism of these changes remains unknown. To further explore this issue, we investigated the changes in the blood supply of the hippocampus in transgenic T2DM mice. In the current study, histochemistry, immunohistochemistry, and unbiased stereological methods were utilized to research the effects of T2DM on hippocampal capillaries of transgenic db/db mice. Twenty (Leprdb) mut/mut mice and twenty (Leprdb) wt/wt mice were used in this study. The learning and memory ability was appraised by Morris water maze test.

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“…The db/db mice were treated with low-dose carvacrol (5 mg/kg) or high-dose carvacrol (10 mg/kg), and it was demonstrated that both doses significantly alleviated the histological abnormalities of the abdominal aorta in the db/db mouse model. It is well known that T2DM-related disorders are often associated with insulin resistance ( 23 - 25 ). Peripheral target tissues exhibit reduced sensitivity to insulin, leading to abnormal insulin secretion and even hyperglycemia.…”

Section: Discussionmentioning

confidence: 99%

Carvacrol may alleviate vascular inflammation in diabetic db/db mice

et al. 2020

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Type 2 diabetes mellitus (T2dM) is associated with chronic low-grade inflammation. carvacrol has been confirmed to possess anti-inflammatory properties, but its effect on diabetic vasculature remains unknown. The aim of the present study was to investigate the possible protective effects of carvacrol against vascular endothelial inflammation. The mice were divided into four groups (n=15 per group) as follows: Non-diabetic control mice, db/db mice, db/db mice + carvacrol (low) and db/db mice + carvacrol (high) groups. The effects of carvacrol on the pathomorphism of the thoracoabdominal aorta in db/db mice were evaluated using hematoxylin and eosin and Masson's trichrome staining. The serum levels of insulin signaling molecules, such as phosphorylated insulin receptor, phosphorylated insulin receptor substrate-1, insulin, triglyceride (TG) and inflammatory cytokines [tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and IL-8] were measured by ELISA. Furthermore, the protein levels of the toll-like receptor (TLR)4/nuclear factor (NF)-κB inflammatory signaling pathway molecules were investigated in the thoracoabdominal aorta of db/db mice and in high glucose-induced endothelial cells. Vascular endothelial cell apoptosis and viability were assessed by using flow cytometry and Cell Counting Kit-8 assays, respectively. The results demonstrated that carvacrol alleviated vascular endothelial cell injury. carvacrol reduced the expression levels of insulin signaling molecules, insulin, TG and inflammatory cytokines in the serum of db/db mice. Moreover, carvacrol reduced the activation of the TLR4/NF-κB signaling pathway in vivo and in vitro. In vitro, carvacrol inhibited high glucose-induced endothelial cell function by promoting vascular endothelial cell apoptosis and suppressing cell viability. These findings demonstrated that carvacrol could alleviate endothelial dysfunction and vascular inflammation in T2DM.

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Changes of Blood-Brain Barrier and Brain Parenchymal Protein Expression Levels of Mice under Different Insulin-Resistance Conditions Induced by High-Fat Diet

Cited by 39 publications

References 39 publications

Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis

Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis

Changes in hippocampal capillaries in transgenic type 2 diabetic mice: A stereological investigation

Carvacrol may alleviate vascular inflammation in diabetic db/db mice

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