Inhibition of N-methyl-N-nitrosourea-induced gastric tumorigenesis by Liuwei Dihuang Pill in db/db mice

Table 1 Compositions of Liuwei Dihuang Pill

Chinese name	Latin name	Parts used	Weight, g
Shu di huang	Rehmannia glutinosa Libosch	Root	160
Shan zhu yu	Cornus officinalis Sieb	Fruit	80
Shan yao	Dioscorea opposite Thunb	Rootstock	80
Ze xie	Alisma orientale (G. Samuelsson) Juz	Rootstock	60
Fu ling	Poria cocos (Schw.) Wolf	Sclerotium	60
Mu dan pi	Paeonia suffruticosa Andrews	Bark	60

Plant name as verified at http://www.theplantlist.org.

Table 2 Sequences of primers for qRT-PCR

Gene	Forward primer (5’-3’)	Reverse primer (5’-3’)
IGF-1	CTGGACCAGAGACCCTTTGC	GGACGGGGACTTCTGAGTCTT
IGF-1R	CATGTGCTGGCAGTATAACCC	TCGGGAGGCTTGTTCTCCT
IGF-2	ACAACTTCGATTTGAACCACATTC	GAGAGCTCAAACCATGCAAACT
IGF-2R	TGAATGGTGATCCTTGCCCTC	CCGGTAGCTGTTGGTCTGTC
Insulin receptor	TCAAGACCAGACCCGAAGATT	TCTCGAAGATAACCAGGG
GAPDH	CCAATGTGTCCGTCGTGGATCT	GTTGAAGTCGCAGGAGACAACC

Table 3 Effects of Liuwei Dihuang Pill on body weights and serum biochemical parameters

Group	Body weights at week 20, g	Body weights at week 30, g	ALT, U/L	AST, U/L	BUN, mmol/L	Cr, μmol/L
db/m + MNU + saline	24.80 ± 1.08	25.56 ± 1.78	8.98 ± 1.78e	8.91 ± 1.45	4.80 ± 0.26	13.94 ± 4.17
db/db + MNU + saline	59.22 ± 5.61	61.18 ± 2.10	53.31 ± 8.31	10.23 ± 0.90	4.65 ± 0.52	18.44 ± 6.28
db/db + MNU + metformin	59.44 ± 3.38	48.90 ± 4.99b	23.40 ± 2.37b	10.79 ± 1.52	4.64 ± 0.51	9.45 ± 3.07
db/db + MNU + LDP	62.79 ± 5.03	57.96 ± 3.34	38.08 ± 4.81a	11.30 ± 1.14	4.85 ± 0.45	9.00 ± 1.50

Data are presented as mean ± SE.

^aP < 0.05,

^bP < 0.01,

^eP < 0.001 vs db/db + MNU + saline group by one-way ANOVA. ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; LDP: Liuwei Dihuang Pill; MNU: N-methyl-N-nitrosourea.

Table 4 Incidence of gastric dysplasia and carcinoma in the experimental mice

Group	Survival	Mortality	Histopathology findings (no. of mice)
			Chronic gastritis	Gastric dysplasia	Gastric carcinoma
db/m + MNU + saline	12	0	5 (42)	2 (17)a	0
db/db + MNU + saline	9	3 (25)	6 (67)	6 (67)	2 (22)
db/db + MNU + metformin	10	2 (17)	6 (60)	1 (10)a	0
db/db + MNU + LDP	10	2 (17)	5 (50)	1 (10)a	0

Data are presented as n (%).

^aP < 0.05 vs incidence of gastric dysplasia in db/db + MNU + saline group. Fisher’s exact test was used for comparison among groups. LDP: Liuwei Dihuang Pill; MNU: N-methyl-N-nitrosourea.