J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Original Research

Volume 2, Number , June 2018, pages 98-103

5-Demethylnobiletin is more effective than nobiletin in preventing AOM/DSS-induced colorectal carcinogenesis in ICR mice

Figure 1.
Structures of (A) nobiletin and (B) 5-demethylnobiletin.

Figure 2.
Experimental design of AOM/DSS-induced colorectal tumorigenesis in male ICR mice for 15 weeks.
Effects of NOB or DMNB prevented AOM/DSS-induced colorectal tumorigenesis.

Figure 3.
Morphology of colonic mucosal surface of AOM/DSS-treated mice.
(A) control group (B) AOM/DSS group (C) NOB and (D) DMNB group. Positive: AOM/DSS.

Figure 4.
Inhibitoy effect of NOB and DMNB on AOM/DSS-induced IL-6 expression and distribution in colorectal tissue.
(A) Colonic tissues were embedded in paraffin and visualized by immunohistochemical (IHC) analysis using IL-6 antibody demonstrated as dark stain. (B) Quantification of IL-6-labeling index (%) was determined by ImageJ (National Institute of Health). *p < 0.05, compared with control group. #p < 0.05, compared with AOM/DSS group.

Figure 5.
The NOB and DMNB significantly prevented AOM/DSS-induced inflammatory responses in colonic mucosa tissue.
Total protein lysates from colonic mucosa detected the expression level of IL-6 (A) and COX-2 (B) by western blot. (C) Reverse transcriptional PCR analyzed colonic mucosa mRNA expression level of IL-6 and COX-2.

**Table 1.** Effects of NOB or DMNB on body weight and relative organ weight in AOM/DSS-induced CRC model
Groups	Body Weight (g)	Relative organ weight (g)
The data of body weight are expressed as the mean ± SD (n = 12) and were statistically analyzed using a one-way ANOVA followed by Duncan’s multiple-range test. Different letters (a−b) represent significant differences among groups, p < 0.05. The data of relative organ weight performed by student’s t-tests, *p < 0.05, compared with control group.
Control	50.7 ± 3.3^a	2.27 ± 0.16	0.85 ± 0.10	0.12 ± 0.04
AOM/DSS	43.3 ± 3.3^b	2.25 ± 0.36	0.84 ± 0.09	0.23 ± 0.09*
AOM/DSS + 0.01% NOB	46.8 ± 4.8^c	2.12 ± 0.31	0.85 ± 0.07	0.13 ± 0.03
AOM/DSS + 0.025% NOB	48.8 ± 3.7^ac	2.16 ± 0.22	0.86 ± 0.10	0.15 ± 0.08
AOM/DSS + 0.01% DMNB	47.6 ± 4.6^ac	2.09 ± 0.26	0.85 ± 0.13	0.13 ± 0.03
AOM/DSS + 0.025% DMNB	48.2 ± 3.2^ac	2.10 ± 0.22	0.84 ± 0.09	0.13 ± 0.05

Table 1. Effects of NOB or DMNB on body weight and relative organ weight in AOM/DSS-induced CRC model

Groups

Body Weight (g)

Relative organ weight (g)

Liver

Kidney

Spleen

The data of body weight are expressed as the mean ± SD (n = 12) and were statistically analyzed using a one-way ANOVA followed by Duncan’s multiple-range test. Different letters (a−b) represent significant differences among groups, p < 0.05. The data of relative organ weight performed by student’s t-tests, *p < 0.05, compared with control group.

Control

50.7 ± 3.3^a

2.27 ± 0.16

0.85 ± 0.10

0.12 ± 0.04

AOM/DSS

43.3 ± 3.3^b

2.25 ± 0.36

0.84 ± 0.09

0.23 ± 0.09*

AOM/DSS + 0.01% NOB

46.8 ± 4.8^c

2.12 ± 0.31

0.85 ± 0.07

0.13 ± 0.03

AOM/DSS + 0.025% NOB

48.8 ± 3.7^ac

2.16 ± 0.22

0.86 ± 0.10

0.15 ± 0.08

AOM/DSS + 0.01% DMNB

47.6 ± 4.6^ac

2.09 ± 0.26

0.85 ± 0.13

0.13 ± 0.03

AOM/DSS + 0.025% DMNB

48.2 ± 3.2^ac

2.10 ± 0.22

0.84 ± 0.09

0.13 ± 0.05

**Table 2.** Effects of NOB or DMNB on the colon length and number of tumors in AOM/DSS-induced CRC model
Groups	Colon length (mm)	NO. of Tumor/colon
The data of body weight are expressed as the mean ± SD (n = 12) and were statistically analyzed using a one-way ANOVA followed by Duncan’s multiple-range test. Different letters (a−b) represent significant differences among groups, p < 0.05.
Control	87.1 ± 5.7^a	1.1 ± 1.1^a	0.0 ± 0.0^a	1.1 ± 1.1^a
AOM/DSS	67.4 ± 6.0^b	10.4 ± 6.2^b	14.0 ± 10.4^b	24.4 ± 8.5^b
AOM/DSS + 0.01% NOB	78.3 ± 14.3^bc	7.8 ± 1.8^ab	2.4 ± 2.8^a	10.2 ± 3.8^c
AOM/DSS + 0.025% NOB	90.6 ± 14.4^ac	7.4 ± 2.1^c	1.4 ± 2.0^a	8.9 ± 2.5^c
AOM/DSS + 0.01% DMNB	81.2 ± 10.2^ac	6.9 ± 2.8^c	1.1 ± 1.4^a	8.0 ± 3.6^c
AOM/DSS + 0.025% DMNB	101.3 ± 11.4^c	6.6 ± 1.9^c	1.0 ± 2.0^a	7.6 ± 3.1^c

Table 2. Effects of NOB or DMNB on the colon length and number of tumors in AOM/DSS-induced CRC model

Groups

Colon length (mm)

NO. of Tumor/colon

Small

Larger

Total

Control

87.1 ± 5.7^a

1.1 ± 1.1^a

0.0 ± 0.0^a

1.1 ± 1.1^a

AOM/DSS

67.4 ± 6.0^b

10.4 ± 6.2^b

14.0 ± 10.4^b

24.4 ± 8.5^b

AOM/DSS + 0.01% NOB

78.3 ± 14.3^bc

7.8 ± 1.8^ab

2.4 ± 2.8^a

10.2 ± 3.8^c

AOM/DSS + 0.025% NOB

90.6 ± 14.4^ac

7.4 ± 2.1^c

1.4 ± 2.0^a

8.9 ± 2.5^c

AOM/DSS + 0.01% DMNB

81.2 ± 10.2^ac

6.9 ± 2.8^c

1.1 ± 1.4^a

8.0 ± 3.6^c

AOM/DSS + 0.025% DMNB

101.3 ± 11.4^c

6.6 ± 1.9^c

1.0 ± 2.0^a

7.6 ± 3.1^c