Comparison of Plasma Exosome Proteomes Between Obese and Non-Obese Patients with Type 2 Diabetes Mellitus

Figures & data

Table 1 Clinical Characteristics of Patients

Variables	Non-Obese DM (n = 10)	Obese DM (n = 10)	P value
Age (years)	49.00± 5.16	69.29 ± 7.91	0.033
Gender
Male (%)	5.00 (50.0)	6.00 (60.0)	0.673
Female (%)	5.00 (50.0)	4.00 (40.0)
Duration (years)	0.99 ± 1.69	1.53 ± 1.40	0.447
Height (cm)	164.70± 7.03	172.40±8.32	0.032
Weight (kg)	63.50 ± 0.04	98.90± 20.17	0.000
BMI (kg/m2)	23.36 ±1.52	33.27 ± 5.45	0.000
ALT (U/L)	21.50± 9.56	77.00± 77.68	0.039
AST (U/L)	16.80±4.44	42.80± 37.39	0.056
TG (mmol/L)	2.38± 1.78	2.35± 1.00	0.967
TC (mmol/L)	5.84 ± 1.00	5.30 ± 0.83	0.206
HDL-C (mmol/L)	1.12± 0.13	0.93 ± 0.16	0.009
LDL-C (mmol/L)	3.23± 1.01	3.26± 0.60	0.941
BUN (mmol/L)	5.92 ±0.90	4.19± 1.00	0.001
Cr (μmol/L)	59.7 ± 17.11	65.90 ± 15.08	0.401
UACR (mg/mmol)	0.37 ± 0.38	0.70 ± 0.51	0.192
eGFR (mL/minl)	107.23 ±9.92	111.45± 18.11	0.529
FPG (mmol/L)	11.01 ± 2.82	9.68± 4.92	0.468
INS (mU/L)	11.11± 5.61	24.28± 11.50	0.004
HbA1C (%)	9.33± 1.62	8.96 ± 2.00	0.655
HOMA-IR	5.52± 3.41	9.33 ± 3.63	0.026
AHI	7.79± 3.48	18.83 ± 8.49	0.003

Abbreviations: DM, diabetes mellitus; BMI, body mass index; ALT, alanine aminotransferase aspartate; AST, aminotransferase; TG, triglyceride; TC, total cholesterol; HDL-C, high density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; BUN, blood urea nitrogen; Cr, serum creatinine; UACR, urine albumin-to-creatinine ratio; eGFR, estimated glomerular filtration rate; FPG, fasting plasma glucose; INS, insulin; HbA1c, hemoglobin A1c; HOMA-IR, homeostasis model assessment of insulin resistance; AHI, the apnea–hypopnea index.

Figure 1 Characterization of plasma exosomes. Plasma exosomes were isolated from obese and non-obese patients with type 2 diabetes. (A) The morphology of plasm exosomes from sample 1 and sample 4 under a transmission electron microscope. Magnification 6000 ×. (B) Particle size was measured using nanoparticle tracking analysis.

Abbreviations: OB, obese patients with type 2 diabetes; DM, non-obese patients with type 2 diabetes.

Table 2 The Differentially Expressed Proteins in Obese Patients with Type 2 Diabetes

Protein	Regulation	Ratio of LFQ (OB/DM)	P value
C9	Up	1.815911592	0.031410504
PON1	Up	1.774121352	0.014398211
HPX	Down	0.734564827	0.045560876
A1BG	Down	0.668644594	0.030692879
CFHR1	Down	0.442142469	0.036910981
ANG	Down	0.649168087	0.015345186
CALM3; CALM2; CALM1	Down	0.437797365	0.010452527

Abbreviations: LFQ, label-free quantification; OB, obese patients with T2D; DM, diabetes mellitus.

Figure 2 Identification of differentially expressed plasma exosome proteins. (A and B). A label-free quantitative mass spectrometry analysis was performed and identified 2 significantly upregulated proteins and 5 significantly downregulated proteins in obese patients with type 2 diabetes. (C). Heatmap of differentially expressed plasma exosome proteins.

Abbreviations: OB, obese patients with type 2 diabetes; DM, non-obese patients with type 2 diabetes.

Figure 3 Comparison of the expression of differentially expressed plasma exosome proteins between obese and non-obese diabetic patients. *p<0.05.

Figure 4 Gene ontology (GO) analysis and Kyoto encyclopedia of genes and genomes (KEGG) analysis. (A) Left: GO enrichment bubble chart. Right: GO enrichment circle diagram. The first circle represents the top 20 GO terms. The second circle represents the number and -log10 (P-value) of the differentially expressed proteins enriched in the corresponding GO term. The third circle indicates the number of upregulated or downregulated proteins. Blue represents upregulated protein. Green represents downregulated proteins. The fourth circle: rich factor value of each GO term. (B) Left: KO enrichment bar chart of top 20 pathways. Right: KO enrichment circle diagram. The first circle is the top 20 enriched pathways. The second circle represents the number of proteins in the corresponding pathway. The third circle indicates the number of upregulated or downregulated proteins. Blue represents upregulated proteins. Green represents downregulated proteins. The fourth circle indicates the rich factor value of each pathway.

Figure 5 Characterization of the differentially expressed proteins. (A) Prediction of the subcellular localization of the differentially expressed proteins. (B) Left: Domain enrichment bubble chart. Right: Domain enrichment circle diagram. The first circle represents the top 20 enriched domains. The second circle reprensents the number of the proteins with the corresponding domain. The third circle represents the number of upregulated or downregulated proteins. Blue represents upregulated proteins. Green represents downregulated proteins. The fourth circle: rich factor value of each domain.

Figure 6 Transcription factor analysis and establishment of the protein-protein interaction network. (A) Transcription factor analysis identifies HMG-box and NF-Y beta may regulate the transcription of the differentially expressed proteins. (B) A protein-protein interaction network. Blue represents upregulated proteins. Green represents downregulated proteins.