Metabolomics Study of Isocaloric Different Dietary Patterns on the Life Span in Healthy Population

Figures & data

Table 1 Recommended Energy Intake Standards for Dietary Nutrients for Chinese Residents (2013 Edition)

Age	Energy (Kcal/d)
	Male	Female
18~	2250	1800
50~	2100	1750
65~	2050	1700

Table 2 Characteristics of Participants in Different Age Groups and Different Isocaloric Diet Groups

Characteristics	Age Groups			Diet Groups
	Young Adult	Middle-Age	Old-Age	NC	IHP	IHF	IHC
	(n=60)	(n=60)	(n=60)	(n=45)	(n=45)	(n=45)	(n=45)
Age (y)	35.08±5.47	53.33±6.71**	68.62±2.79**^##	52.38±14.97	52.33±12.86	51.6±14.61	52.27±14.95
BMI (kg/m^2)	23.98±4.01	24.76±3.66	24.89±3.50	24.28±5.17	24.48±2.88	24.45±2.97	24.97±3.54
GLU (mmol/L)	4.23±0.66	4.93±1.67*	5.3±2.00**	4.82±1.96	5.11±2.17	4.74±0.97	4.62±0.94
UA (μmol/L)	277.29±74.07	275.21±79.57	296.85±77.44	279.33±78.32	277.29±75.82	292.42±87.66	283.43±67.61
Ca (mmol/L)	2.23±0.20	2.24±0.10	2.27±0.11	2.24±0.09	2.24±0.11	2.26±0.12	2.23±0.22
P (mmol/L)	1.11±0.20	1.11±0.16	1.15±0.66	1.09±0.16	1.07±0.20	1.23±0.75	1.10±0.16
TCHO (mmol/L)	4.57±0.91	5.05±0.77**	5.46±0.78**^#	4.68±0.71	5.33±0.97^ΔΔ	4.97±0.85	5.14±0.93^Δ
TG (mmol/L)	0.96±0.62	1.16±0.78	1.50±0.76**^#	0.99±0.46	1.39±0.91^Δ	1.12±0.65	1.32±0.86
HDL-C (mmol/L)	1.33±0.27	1.31±0.28	1.21±0.28	1.38±0.30	1.23±0.29^Δ	1.27±0.27	1.27±0.26
LDL-C (mmol/L)	2.65±0.77	2.94±0.65*	3.29±0.71**^#	2.55±0.61	3.23±0.77^ΔΔ	2.92±0.70^ΔΔ	3.14±0.76^ΔΔ
Dietary intake
Total energy intake (kcal/d)	2134.63±530.03	2014.28±285.56	2156.01±560.13	2154.23±465.76	2219.07±608.49	2010.38±467.61	2022.87±291.61
Fat (% of energy)	31.77±7.66	30.00±6.57	29.99±7.67	28.31±1.13	33.41±6.95^ΔΔ	38.39±4.24^ΔΔ	22.23±1.87^ΔΔ
Protein (% of energy)	13.38±3.62	12.85±2.71	12.93±2.60	12.37±1.09	17.10±2.72^ΔΔ	12.21±1.61	10.52±1.08^ΔΔ
Carbohydrate (% of energy)	57.73±9.74	59.58±8.59	60.01±9.10	62.30±1.61	52.67±81^ΔΔ	51.32±1.51^ΔΔ	70.13±2.19^ΔΔ

Notes: *P<0.05, **P<0.01 vs young adult group. ^#P<0.05, ^##P<0.01 vs middle-age group. ^ΔP<0.05, ^ΔΔP<0.01 vs normal control group. All values are mean ± SD. One-way ANOVA followed by Dunnett post hoc test.

Abbreviations: BMI, the body mass index; Ca, blood calcium; IHC, isocaloric high-carbohydrate group; IHF, isocaloric high-fat group; IHP, isocaloric high-protein group; NC, normal control group; P, blood phosphorus; UA, blood uric acid.

Figure 1 PCA scores plot and validation plot of permutation testing of PLS-DA model. (A and B) PCA scores plot generated from serum samples of participants and QC in positive and negative mode. Purple triangle, QC. Light blue diamond, Sample. (C and D) The validation plot of permutation testing of PLS-DA model. (E and F) PLS-DA score plots generated from serum samples of participants in different age groups. Green dot, young group. Blue box, middle-aged group. Crimson triangle, old-aged group. (A, C and E) the positive mode. (B, D and F) the negative mode.

Figure 2 PLS-DA score plots generated from serum samples of participants in different diet groups. (A–C) The positive ESI mode. (D–F) The negative ESI mode.

Table 3 Ionic Strength of 18 Potential Life-Related Biomarkers in Different Age Groups and Different Isocaloric Diet Groups

Metabolites (μmol/L)	Age Groups			Diet Groups
	Young Adult	Middle-Age	Old-Age	NC	IHP	IHF	IHC
Methionine sulfoxide	9.44±1.91	6.79±1.77**	0.60±1.71**^##	6.69±4.35	4.81±3.71^Δ	6.10±4.46	4.82±3.70^Δ
LysoPC (O-18:0)	11.13±3.71	5.95±3.57**	0.06±0.47**^##	7.03±5.96	4.61±4.79^Δ	6.54±5.84	4.63±4.68^Δ
LysoPC (20:4)	26.14±17.63	3.18±7.96**	0.23±0.68**	13.44±18.59	6.46±13.73^Δ	11.87±16.67	7.49±14.16
LysoPC (20:1)	14.77±4.78	10.56±2.86**	3.92±5.56**^##	11.65±5.68	8.05±6.56^ΔΔ	10.34±4.89	8.90±7.63^Δ
LysoPE (18:2/0:0)	13.12±4.29	0.69±1.56**	0.22±0.61**	5.30±7.16	3.88±5.79	5.62±7.58	3.59±5.56
γ-Aminobutyric acid	16.75±3.31	10.26±7.44**	3.60±5.67**^##	11.94±7.94	8.61±7.64^Δ	12.06±7.18	8.15±7.97^Δ
L-Valine	1.66±0.96	0.91±0.55**	0.02±0.72**^##	1.11±1.19	0.68±0.71^Δ	0.97±1.00	0.66±0.69^Δ
L-Threonine	54.22±8.49	21.28±23.64**	1.38±1.91**^##	32.03±27.09	21.09±25.33	27.15±27.65	20.15±25.38^Δ
Aminoadipic acid	0.02±0.14	1.08±1.60**	3.23±1.70**^##	1.00±1.51	1.81±2.25^Δ	1.12±1.55	1.86±2.11^Δ
Leukotriene F4	0.01±0.07	0.48±0.54	2.32±2.35**^##	0.54±0.61	1.43±2.27^Δ	0.56±0.66	1.26±2.33^Δ
Taurocholic acid	0.93±0.72	0.76±0.88	0.26±0.53**^##	0.88±0.87	0.45±0.65^ΔΔ	0.74±0.80	0.53±0.72^Δ
Serotonin	2.26±1.77	1.12±1.15**	0.80±1.31**	1.65±1.58	0.82±1.22^ΔΔ	2.25±1.64^Δ	0.84±1.17^ΔΔ
Succinic acid	0.07±0.25	0.14±0.30	3.46±2.28**^##	0.69±1.12	1.56±2.36^Δ	0.71±0.87	1.95±2.97^ΔΔ
Glycocholic acid	1.13±0.90	0.07±0.15**	0.04±0.12**	0.54±0.89	0.22±0.37^Δ	0.66±0.98	0.25±0.40
cis-Aconitic acid	0.07±0.21	0.76±1.40	12.20±10.71**^##	2.47±4.44	6.31±9.71^Δ	2.22±3.51	6.45±11.96^Δ
9,10- DiHODE	2.34±1.33	0.24±0.94**	0.15±0.73**	1.27±1.68	0.65±1.11^Δ	1.10±1.63	0.63±1.20^Δ
12,13- DHOME	0.12±0.38	0.21±0.55	1.67±1.08**^##	0.42±0.77	0.85±1.12^Δ	0.56±0.90	0.84±1.22^Δ
Deoxycholic acid	6.79±7.95	9.54±8.97	14.65±12.00**^#	7.84±8.53	12.85±10.43^Δ	8.04±8.56	12.68±12.35^Δ

Notes: **P<0.01 vs young adult group. ^#P<0.05, ^##P<0.01 vs middle-age group. ^ΔP<0.05, ^ΔΔP<0.01 vs normal control group. All values are mean ± SD. One-way ANOVA followed by Dunnett post hoc test.

Abbreviations: IHC, isocaloric high-carbohydrate group; IHF, isocaloric high-fat group; IHP, isocaloric high-protein group; NC, normal control group.

Figure 3 ROC curves of 18 potential biomarkers in young adult group vs old-age group. 18 potential biomarkers include LysoPC (20:1) (AUC=0.923), γ-aminobutyric acid (AUC=0.971), L-valine (AUC=0.983), L-threonine (AUC=1), aminoadipic acid (AUC=0.939), leukotriene F4 (AUC=0.965), taurocholic acid (AUC=0.746), serotonin (AUC=0.738), LysoPE (18:2) (AUC=1), succinic acid (AUC=0.979), glycocholic acid (AUC=0.982), cis-Aconitic acid (AUC=0.973), methionine sulfoxide (AUC=0.998), LysoPC (O-18:0) (AUC=1), LysoPC (20:4) (AUC=0.837), 9,10-DiHODE (AUC=0.92), 12,13- DHOME (AUC=0.948), and deoxycholic acid (AUC=0.7130).

Figure 4 One-way ANOVA followed by Dunnett post hoc test of different age groups and diet groups. (A) Comparison of amino acids concentrations in different age groups. (B) Comparison of amino acids concentration in different isocaloric dietary groups. (C) Comparison of free fatty acids concentrations in different age groups. (D) Comparison of free fatty acids concentrations in different isocaloric dietary groups. *P<0.05, **P<0.01 vs young adult group. ^#P<0.05 vs middle-age group. ^▲P<0.05, ^▲▲P<0.01 vs normal control group.

Abbreviations: NC, normal control group; IHP, isocaloric high-protein group; IHF, isocaloric high-fat group; IHC, isocaloric high-carbohydrate group.