Differentially expressed plasma proteins of β-thalassemia/hemoglobin E patients in response to curcuminoids/vitamin E antioxidant cocktails

Figures & data

Table 1. Clinical data of the patients recruited in this study.

Parameters	Curcuminoids cocktail (n = 10)			Vitamin E cocktail (n = 10)
	Baseline	Month 6	Month 12	Baseline	Month 6	Month 12
Hematological parameter
Hemoglobin (g/l)	65.1 ± 3.1	69.0 ± 2.4*	70.3 ± 3.0*	71.0 ± 5.5	74.0 ± 5.2*	77.0 ± 5.8*
Iron load parameters
Serum ferritin (pmol/l)	2520 ± 542	2126 ± 530*	1969 ± 395*	4496 ± 1061	2201 ± 339*	2009 ± 375*
Serum NTBI (µmol/l)	6.4 ± 1.5	4.4 ± 1.0	2.3 ± 1.0*	5.2 ± 0.8	4.1 ± 1.0	2.7 ± 0.8*
Oxidative stress parameters
Blood ROS (MFC)	27.0 ± 2.6	22.3 ± 2.8	19.1 ± 2.1*	32.9 ± 7.2	22.4 ± 4.4	16.9 ± 2.3*
RBC MDA (nmol/g Hb)	983 ± 34	849 ± 66*	742 ± 44**	1354 ± 173	700 ± 27*	672 ± 25*
Procoagulation parameters
PF3-like activity (A405)	0.88 ± 0.08	0.80 ± 0.08	0.69 ± 0.07*	0.97 ± 0.06	0.60 ± 0.07*	0.44 ± 0.05**
PS+ RBC (%)	5.71 ± 1.83	1.07 ± 0.34*	0.49 ± 0.15*	4.90 ± 1.37	2.21 ± 1.02	0.34 ± 0.08*

Notes: MDA: malondialdehyde; NTBI: non-transferrin-bound iron; PF3: platelet factor 3; PS+: phosphatidylserine-positive; RBC: red blood cells; ROS: reactive oxygen species.

* p < .05 compared to baseline; ** p < .001 compared to baseline

Table 2. Plasma proteins differentially expressed in β-thalassemia/Hb E patients treated with curcuminoids antioxidant cocktail.

Protein	MW (kDa)/pI	MS/MS score	Peptide matches	%Coverage	%volume protein intensity
					Normal	Baseline	Month 6	Month 12
Adhesive protein
Vitronectin	55.07/5.55	184	3	8	1.007 ± 0.110	0.686 ± 0.046^a	0.913 ± 0.055^d	0.758 ± 0.071
Vitronectin fragment 1	55.07/5.55	101	2	5	0.375 ± 0.044	0.223 ± 0.016^a	0.243 ± 0.019	0.280 ± 0.028^b
Antioxidants
Paraoxonase/ arylesterase 1	39.88/5.08	82	3	7	0.234 ± 0.030	0.153 ± 0.006^a	0.161 ± 0.010	0.213 ± 0.021^b
Protein AMBP	39.89/5.95	85	1	3	0.027 ± 0.004	0.012 ± 0.001^a	0.012 ± 0.001	0.016 ± 0.002^d
Cell motility; actin-binding proteins
Myosin-1 fragment 1	223.98/5.59	76	2	1	0.203 ± 0.013	0.159 ± 0.013^a	0.212 ± 0.019^b	0.216 ± 0.023^b
Tropomyosin alpha-3 chain	28.91/4.72	103	4	12	0.062 ± 0.014	0.008 ± 0.001^a	0.017 ± 0.003^b	0.024 ± 0.006^b
Lipoproteins
Apolipoprotein E isoform 1	36.25/5.65	149	10	25	0.148 ± 0.030	0.284 ± 0.013^a	0.233 ± 0.017^b	0.207 ± 0.020^b
Apolipoprotein E isoform 2	36.25/5.65	177	11	28	0.255 ± 0.041	0.519 ± 0.019^c	0.442 ± 0.022^b	0.394 ± 0.041^b
Apolipoprotein A-I fragment 1	30.76/5.56	85	1	4	0.023 ± 0.008	0.006 ± 0.001^a	0.012 ± 0.003^b	0.009 ± 0.001^b
Apolipoprotein C-II	8.91/4.66	172	4	73	0.142 ± 0.025	0.214 ± 0.020^a	0.166 ± 0.016^b	0.155 ± 0.013^b
Immune system proteins
Complement factor H fragment 1	143.68/6.21	318	11	11	0.069 ± 0.007	0.049 ± 0.004^a	0.066 ± 0.007^b	0.062 ± 0.006^b
C4b-binding protein alpha chain fragment 1	69.04/7.15	83	2	4	0.024 ± 0.003	0.014 ± 0.001^a	0.023 ± 0.003^b	0.026 ± 0.002^b
Coagulation proteins
Prothrombin fragment 1	71.48/5.64	148	5	9	0.229 ± 0.032	0.079 ± 0.007^c	0.086 ± 0.009	0.102 ± 0.012^b
Fibrinogen gamma chain fragment 1	52.11/5.37	173	3	7	0.056 ± 0.022	0.007 ± 0.001^a	0.011 ± 0.002^b	0.013 ± 0.002^b
Carrier protein
Transthyretin isoform 1	15.99/5.52	81	2	23	0.031 ± 0.005	0.092 ± 0.008^c	0.121 ± 0.011^b	0.069 ± 0.005^b
Transthyretin isoform 2	15.99/5.52	131	2	23	0.380 ± 0.023	0.772 ± 0.026^c	0.685 ± 0.034^b	0.594 ± 0.035^b

Note: Values were mean ± SEM. ^a Compared to normal, p < .05; ^b Compared to baseline, p < .05; ^c Compared to normal, p < .001; ^d Compared to baseline, p < .001.

Table 3. Plasma proteins differentially expressed in β-thalassemia/Hb E patients treated with vitamin E antioxidant cocktail.

Protein	MW (kDa)/pI	MS/MS score	Peptide matches	%Coverage	%volume protein intensity
					Normal	Baseline	Month 6	Month 12
Cell motility; actin-binding proteins
Gelsolin	86.04/5.90	74	1	1	0.019 ± 0.004	0.007 ± 0.001^a	0.007 ± 0.002	0.010 ± 0.001^b
Immune system proteins
Complement C3 fragment 1	188.6/6.02	308	6	18	0.003 ± 0.001	0.036 ± 0.002^c	0.033 ± 0.003	0.021 ± 0.003^b
Complement C3 fragment 2	188.6/6.02	127	5	2	0.058 ± 0.009	0.391 ± 0.017^c	0.320 ± 0.016^b	0.268 ± 0.010^d
Complement factor H fragment 1	143.68/6.21	318	11	11	0.069 ± 0.007	0.046 ± 0.004^a	0.057 ± 0.005^b	0.063 ± 0.008^b
Inflammatory response protein
CD5 antigen-like	39.60/5.28	133	2	7	0.262 ± 0.035	0.564 ± 0.053^c	0.495 ± 0.045^b	0.485 ± 0.046^b
Lipoproteins
Apolipoprotein A-IV	45.37/5.28	57	1	3	0.079 ± 0.009	0.048 ± 0.006^a	0.061 ± 0.009	0.093 ± 0.010^d
Apolipoprotein E isoform 2	36.25/5.65	177	11	28	0.255 ± 0.041	0.523 ± 0.038^c	0.477 ± 0.036	0.395 ± 0.030^b
Apolipoprotein A-I fragment 1	30.76/5.56	85	1	4	0.023 ± 0.008	0.005 ± 0.001^a	0.007 ± 0.001^b	0.009 ± 0.002^b
Coagulation proteins
Fibrinogen gamma chain fragment 1	52.11/5.37	191	4	12	0.046± 0.004	0.019 ± 0.003^c	0.024 ± 0.005	0.033 ± 0.006^b
Fibrinogen gamma chain fragment 2	52.11/5.37	61	1	3	0.020 ± 0.003	0.008 ± 0.001^a	0.010 ± 0.001	0.012 ± 0.001^d
Antioxidants
Glutathione peroxidase 3	25.76/8.26	119	2	11	0.150 ± 0.009	0.176 ± 0.006^a	0.149 ± 0.008^b	0.155 ± 0.006^b
Haptoglobin-related protein fragment 1	39.52/6.63	143	2	3	0.015 ± 0.004	0.004 ± 0.001^a	0.005 ± 0.001^b	0.004 ± 0.001
Adhesive protein
Vitronectin fragment 1	55.07/5.55	101	2	5	0.375 ± 0.044	0.226 ± 0.029^a	0.291 ± 0.033^b	0.261 ± 0.031^b
Vitronectin fragment 2	55.07/5.55	35	1	2	0.018 ± 0.003	0.010 ± 0.001^a	0.017 ± 0.002^b	0.018 ± 0.003^b

Note: Values were mean ± SEM. ^a Compared to normal, p < .05; ^b Compared to baseline, p < .05; ^c Compared to normal, p < .001; ^d Compared to baseline, p < .001.

Figure 1. (A) Correlations between vitronectin intensities and total bilirubin levels; (B) The ELISA results of vitronectin in normal subjects and patients treated with curcuminoids and vitamin E antioxidant cocktails. * p < .001 compared to normal; ** p < .05 compared to normal; ^a p < .001 compared to baseline; ^b p < .05 compared to baseline.

Figure 2. (A) Correlations between paraoxonase/arylesterase 1 intensities and blood reactive oxygen species; (B) The ELISA results of paraoxonase/arylesterase 1 in normal subjects and patients treated with curcuminoids antioxidant cocktail. * p < .001 compared to normal; ** p < .05 compared to normal; ^a p < .05 compared to baseline.

Figure 3. (A) The bands of Western blots corresponding to prothrombin, apolipoprotein E, and apolipoprotein A-1 of normal subjects (N), thalassemic patients at baseline, month 6 (M6), and month 12 (M12) after curcuminoids- and vitamin E antioxidant cocktails treatments. The graphs (B) and (C) represent the quantitation of the Western blots for these proteins as percentage of the mean value for normal subjects in curcuminoids- and vitamin E antioxidant cocktails groups, respectively. Values were mean ± SEM. * p < .05 and ** p < .01 compared to baseline.