Proteomic analysis of serum biomarkers for prediabetes using the Long-Evans Agouti rat, a spontaneous animal model of type 2 diabetes mellitus.
OBJECTIVE
To identify candidate serum molecules associated with the progression of type 2 diabetes mellitus, differential serum proteomic analysis was carried out on a spontaneous animal model of type 2 diabetes mellitus without obesity, the Long-Evans Agouti (LEA) rat.


METHODS
We carried out quantitative proteomic analysis using serum samples from 8- and 16-week-old LEA and control Brown Norway (BN) rats (n = 4/group). Differentially expressed proteins were validated by multiple reaction monitoring analysis using the sera collected from 8-, 16-, and 24-week-old LEA (n = 4/each group) and BN rats (n = 5/each group). Among the validated proteins, we also examined the possible relevance of the human homolog of serine protease inhibitor A3 (SERPINA3) to type 2 diabetes mellitus.


RESULTS
The use of 2-D fluorescence difference gel electrophoresis analysis and the following liquid chromatography-multiple reaction monitoring analysis showed that the serum levels of five proteins were differentially changed between LEA rats and BN rats at all three time-points examined. Among the five proteins, SERPINA3N was increased significantly in the sera of LEA rats compared with age-matched BN rats. The serum level of SERPINA3 was also found to be significantly higher in type 2 diabetes mellitus patients than in healthy control participants. Furthermore, glycated hemoglobin, fasting insulin and estimated glomerular filtration rate were independently associated with the SERPINA3 levels.


CONCLUSIONS
These findings suggest a possible role for SERPINA3 in the development of the early stages of type 2 diabetes mellitus, although further replication studies and functional investigations regarding their role are required.
Department of Diabetic Complications, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Eri
E
Takahashi
Department of Diabetic Complications, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Hiroyuki
http://orcid.org/0000-0003-4827-8805
H
Unoki-Kubota
Department of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Yukiko
Y
Shimizu
Department of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.|Section of Animal Models, Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Tadashi
T
Okamura
Department of Diabetic Complications, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Wakiko
W
Iwata
Department of Diabetes, Endocrinology and Metabolism, Center Hospital, National Center for Global Health and Medicine, Tokyo, Japan.
Hiroshi
H
Kajio
Department of Diabetes, Endocrinology and Metabolism, Center Hospital, National Center for Global Health and Medicine, Tokyo, Japan.
Ritsuko
R
Yamamoto-Honda
Department of Complete Medical Checkup, Center Hospital, National Center for Global Health and Medicine, Tokyo, Japan.
Tomoko
T
Shiga
Department of Diabetes and Endocrinology, JR Tokyo General Hospital, Tokyo, Japan.
Shigeo
S
Yamashita
The First Department of Internal Medicine, Faculty of Medicine, University of Toyama, Toyama, Japan.
Kazuyuki
K
Tobe
Department of Diabetic Complications, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Akinori
A
Okumura
Department of Molecular Metabolic Regulation, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Michihiro
M
Matsumoto
Department of Metabolic Disorder, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Kazuki
K
Yasuda
Department of Endocrinology and Diabetes, Saitama Medical University, Saitama, Japan.
Mitsuhiko
M
Noda
Department of Diabetic Complications, Diabetes Research Center, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan.
Yasushi
Y
Kaburagi
D000209:Acute-Phase Proteins; D000368:Aged; D000818:Animals; D015415:Biomarkers; D003924:Diabetes Mellitus, Type 2 / Q000097:blood*; D004195:Disease Models, Animal; D018450:Disease Progression; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011236:Prediabetic State / Q000097:blood*; D040901:Proteomics; D011919:Rats, Inbred Strains; D020318:Rats, Long-Evans; D015843:Serpins / Q000097:blood
D000209:Acute-Phase Proteins; D015415:Biomarkers; C403317:Serpina3n protein, rat; D015843:Serpins
Long-Evans Agouti rat; Quantitative serum proteomics; Serine protease inhibitor A3