Metabonomics of d-glucaro-1,4-lactone in preventing diethylnitrosamine-induced liver cancer in rats

Figures & data

Table 1. Alteration of rat weight before and after DEN treatment (Mean ± S.E.).

Group	Sex	Weight before treatment (g)	Weight on the 20th week, (g)
Control group	Female	206.25 ± 7.2 (n = 5)	330.23 ± 12.4 (n = 5)
	Male	220.62 ± 12.5 ( n = 5)	505.53 ± 22.1 (n = 5)
DEN group	Female	216.30 ± 8.3 (n = 10)	170.23 ± 13.2 (n = 3)
	Male	217.48 ± 13.4 (n = 10)	306.60 ± 15.5 (n = 6)
DEN + 1,4-GL group	Female	119.00 ± 11.6 (n = 13)	213.17 ± 10.6 (n = 8)
	Male	208.11 ± 11.2 (n = 13)	360.36 ± 13.6 (n = 10)

Table 2. Survival and cancer induction rate of DEN-induced HCC rats.

Group	Sex	Number of rats (before treatment)	Number of surviving rats (20th week )	Survival rate (%)	HCC induction rate (%)
Control group	Female	5	5	100	0
	Male	5	5
DEN group	Female	10	3	45	100
	Male	10	6
DEN + 1,4-GL group	Female	13	8	70	100
	Male	13	10

Figure 1. Photographs of the livers of healthy (a), DEN-induced HCC rat (b) and HCC rat administered 1,4-GL (c); Hepatic pathological changes (HE ×100) in healthy rat (d), DEN-induced HCC rat (e), and HCC rat administered 1,4-GL (f).

Figure 2. The Scores plot of PCA (a) and PLS-DA (b) by ¹HNMR spectra of rat serum (1: Control group, 2: DEN group, 3: DEN + 1,4-GL group).

Table 3. Information of rat serum metabolites for classification observed by ¹HNMR spectra.

Metabolite	δ^1H	VIP	Integral interval	Relative levels in serum (mean ± SE.)
				Control Group	DEN Group (n =	DEN + 1,4GL Group
				(n = 10)	9)	(n = 18)
Leucine	0.94 (d)	2.28	0.95–0.98	2.97 ± 0.01	2.87 ± 0.02*	2.90 ± 0.03
Isoleucine	1.02 (d)	2.15	0.98–1.00	2.58 ± 0.02	2.48 ± 0.02*	2.50 ± 0.02
Valine	1.04 (d)	2.41	1.03–1.05	2.79 ± 0.01	2.65 ± 0.01*	2.70 ± 0.02#
3-Hydroxybutyrate	1.20 (dd)	3.01	1.19–1.22	3.30 ± 0.03	2.89 ± 0.02*	3.09 ± 0.04#
Lactate	1.32 (d)	2.67	1.32–1.35	4.54 ± 0.03	4.06 ± 0.03*	4.31 ± 0.03#
Acetate	1.91 (s)	2.30	1.92–1.93	2.39 ± 0.02	2.59 ± 0.03*	2.44 ± 0.02#
Glutamate	2.34 (m)	1.89	2.31–2.32	3.15 ± 0.03	3.43 ± 0.03*	3.40 ± 0.03
Glutamine	2.44 (m)	2.27	2.44–2.46	2.53 ± 0.02	2.35 ± 0.03*	2.45 ± 0.02#
Citrate	2.55 (d)	2.81	2.64–2.66	2.08 ± 0.03	2.16 ± 0.03*	2.21 ± 0.02#
	2.65 (d)
Glycine	3.57 (s)	2.21	3.56–3.58	2.69 ± 0.02	2.66 ± 0.02*	2.78 ± 0.02#
Tyrosine	6.88 (dd)	2.10	6.89–6.92	2.88 ± 0.02	2.96 ± 0.04*	2.99 ± 0.04
	7.18 (dd)
Formate	8.45 (s)	1.55	8.45–8.47	2.03 ± 0.03	2.14 ± 0.02*	2.10 ± 0.03

VIP: variable importance for classification.

* Significantly different (p < 0.05) from the Control group.

# Significantly different (p < 0.05) from the DEN group.

Figure 3. Typical HPLC-UV chromatogram (254 nm) of rat serum after PITC derivatization. Glu: glutamate; Ser: serine; Gly: glycine; His: histidine; Arg: arginine; Ala: alanine; Ty: tyrosine; Val: valine; Ile: isoleucine; Leu: leucine; Nle: norleucine; Phe: Phenylalanine; PITC: phenylisothiocyanate.