Proteomic research of human retinal microvascular pericytes stimulated with high glucose

Authors:Xiao Mengran,  Zhang Xiaomin,  Shao Xianfeng,  Xiao Jing,  Yang Fuhua,  Zhang Hui,  Ea Vicki L,  Li Xiaorong
DOI: 10.3760/cma.j.issn.2095-0160.2019.09.004
Published 2019-09-10
Cite as Chin J Exp Ophthalmol, 2019,37(9): 707-712.

Abstract

Objective

To determine the changes of protein expressions in human retinal microvascular pericytes (HRMPCs) stimulated with high glucose by using quantitative proteomics, which provides new clues for future investigation of diabetic retinopathy (DR).

Methods

HRMPCs were divided into two groups.The cells in control group were cultured in DMEM basic medium with 25 mmol/L glucose, while the cells in high glucose group were cultured in DMEM medium with 35 mmol/L glucose.The amount of living cells was measured by cell counting kit-8(CCK-8). The proteins were collected from the two groups and then were digested with trypsin.Peptides of 2 μg were injected into the time of flight-mass spectrometer and the acquisition mode was DDA.The results were further analyzed using bioinformatics software.

Results

CCK-8 results showed that the absorbance (A450) of HRMPCs in high glucose group was 0.75±0.04, which was significantly lower than 0.91±0.05 in control group (t=5.784, P=0.000 2). In total, 1 972 proteins were identified and 54 of them were significantly different between the two groups (fold change >1.5). Among them, 13 proteins were up-regulated, including CTNNB1 and CTBP2; while 41 proteins were down-regulated, including SQSTM1 and HMGCS1.The differentially expressed proteins were mainly involved in citric acid cycle and aerobic respiration.

Conclusions

The expressions of many proteins in HRMPCs change under the stimulation of high glucose, which may influence the respiration and the ATP production of cells and eventually induce the loss of pericyte.

Key words:

Diabetic retinopathy; Proteomics; Human retinal microvascular pericyte

Contributor Information

Xiao Mengran
Tianjin Key Laboratory of Retinal Functions and Diseases, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin 300384, China
Zhang Xiaomin
Shao Xianfeng
Xiao Jing
Yang Fuhua
Zhang Hui
Ea Vicki L
Li Xiaorong
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Updated: June 24, 2020 — 11:32 am