Brewers’ rice modulates oxidative stress in azoxymethane-mediated colon carcinogenesis in rats

Table 1 Composition of experimental diets

Ingredients (g/1000 g diet)	Group
	G1	G2	G3	G4	G5
Brewers’ rice	-	-	100.0	200.0	400.0
Corn starch	397.5	397.5	315.3	233.2	68.9
Casein	200.0	200.0	191.0	182.0	164.0
Maltodextrin	132.0	132.0	132.0	132.0	132.0
Sucrose	100.0	100.0	100.0	100.0	100.0
Soybean oil	70.0	70.0	68.1	66.1	62.2
Powdered cellulose	50.0	50.0	44.7	39.4	28.7
AIN-93G mineral mix	35.0	35.0	33.4	31.9	28.8
AIN-93G vitamin mix	10.0	10.0	10.0	10.0	10.0
L-cystine	3.0	3.0	3.0	3.0	3.0
Choline bitartrate	2.5	2.5	2.5	2.5	2.5
tert-butylhydroquinone	0.014	0.014	0.014	0.014	0.014

G1 and G2: AIN-93G diet; G3: AIN-93G diet containing 10% (w/w) of brewers’ rice; G4: AIN-93G diet containing 20% (w/w) of brewers’ rice; G5: AIN-93G diet containing 40% (w/w) of brewers’ rice.

Table 2 Nucleotide sequence of polymerase chain reaction primers (obtained from GenBank database)

Primer name[Accession number]	Oligonucleotides (5’-3’)Sequence
GSK3β	F: GGGCACCAGAGCTGATCTTT
[NM_032080]	R: GCCGAAAGACCTTCGTCCA
Beta-catenin	F: CGTGGAAGCTGGTGGGATG
[AF121265]	R: TTCCTGCTTAGTCGCTGCAT
NF-κB	F: AGAGGATGTGGGGTTTCAGG
[NM_001276711.1]	R: GCTGAGCATGAAGGTGGATG
iNOS	F: GTACCCTCAGTTCTGTGCCT
[NM_012611.3]	R: TGTTGCGTTGGAAGTGTAGC
Nrf2	F: TCTGACTCCGGCATTTCACT
[NM_031789.2]	R: CCCCAGAAGAATGTGTTGGC
HO-1	F: CTAGAGCAGGACATGGCCTT
[NM_012580.2]	R: GCCTTCTGCGCAATCTTCTT
ACTB1	F: CCACCCGCGAGTACAACC
[NM_031144.3]	R: TCAGGATGCCTCTCTTGCTC
B2M1	F: CCCACCCTCATGGCTACTTC
[NM_012512.2]	R: GATGAAAACCGCACACAGGC
RPLP11	F: CAAGGTGCTCGGTCCTTCC
[NM_001007604.2]	R: GAGCCTTTGCAAACAAGCCA

¹Housekeeping gene. ACTB: β-actin; B2M: Beta-2 microglobulin; GSK3β: Glycogen synthase kinase 3β; HO-1: Heme oxygenase-1; iNOS: Nitric oxide synthase, inducible; NF-κB: Nuclear factor-kappa B; Nrf2: Nuclear factor E2-related factor 2; RPLP1: Ribosomal protein, large, P1.

Table 3 Colon superoxide dismutase, malondialdehyde, and nitric oxide levels in azoxymethane-induced colon cancer after twenty weeks treatment

Group	SOD (U/mg of protein)	MDA (nmol/g of protein)	NO (μmol/mg of protein)
Normal	60.93 ± 5.23a	5.36 ± 0.23a	414.64 ± 11.59a
AOM alone	43.43 ± 2.96	18.01 ± 1.43	798.46 ± 30.45
AOM + 10% (w/w) of brewers’ rice	43.85 ± 6.32	17.34 ± 3.16	622.70 ± 15.62a
AOM + 20% (w/w) of brewers’ rice	61.71 ± 2.36a	14.24 ± 0.58a	533.40 ± 40.43a
AOM + 40% (w/w) of brewers’ rice	61.29 ± 4.32a	8.14 ± 1.42a	619.35 ± 15.04a

Each value expressed as mean ± SD (n = 3). ^aP < 0.05 vs AOM alone, Tukey’s test. MDA: Malondialdehyde; NO: Nitric oxide; SOD: Superoxide dismutase.