Effects of lymphocyte DNA damage levels in Korean plant food groups and Korean diet regarding to glutathione S-transferase M1 and T1 polymorphisms

Hyun-A Kim; Min-Young Lee; Myung-Hee Kang

doi:10.4163/jnh.2017.50.1.10

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Kim, Lee, and Kang: Effects of lymphocyte DNA damage levels in Korean plant food groups and Korean diet regarding to glutathione S-transferase M1 and T1 polymorphisms

Research Article

J Nutr Health 2017;50(1):10-24.

Published online: 20 January 2017

DOI: https://doi.org/10.4163/jnh.2017.50.1.10

Effects of lymphocyte DNA damage levels in Korean plant food groups and Korean diet regarding to glutathione S-transferase M1 and T1 polymorphisms

Hyun-A Kim, Min-Young Lee, Myung-Hee Kang^†

Department of Food & Nutrition, Daedeok Valley Campus, Hannam University, Daejeon 34054, Korea

^†To whom correspondence should be addressed. tel: +82-42-629-8791, E-mail: mhkang@hnu.kr

Received 13 January 2017 Revised 6 February 2017 Accepted 10 February 2017

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

ABSTRACT Purpose: GST (glutathione S-transferase) M1 and T1 gene polymorphisms are known to affect antioxidant levels. This study was carried out to evaluate genetic susceptibility by measuring the effect of DNA damage reduction in the Korean diet by vegetable food according to GST gene polymorphisms using the ex vivo method with human lymphocytes. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups. A total of 84 foods, which constituted more than 1% of the total intake in each food group, were finally designated as a vegetable food in the Korean diet. The Korean diet applied in this study is the standard one-week meals for Koreans (2,000 Kcal/day) suggested by the 2010 Dietary Reference Intakes for Koreans. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: In the Korean food group, the DNA damage protective effect of GSTM1 and GSTT1 was found to be greater in mutant type and wild-type, respectively. and the DNA damage protective effect according to the combined genotype of GSTM1 and GSTT1 was different depending on the food group. On the other hand, in Korean Diet, the DNA damage protective effect appeared to be larger in GSTM1 wild-type than in mutant type and was found to not be affected by GSTT1 genotype. Conclusion: These results can be used as basic data to demonstrate the superiority of the antioxidant function of Korean dietary patterns and food groups. Furthermore, it may be a starting point to begin research on customized antioxidant nutrition according to individual genes.

Keywords: DNA damage, GST polymorphism, Korean plant food groups, Korean Diet, Comet assay

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Fig. 1.

Relative score of lymphocyte DNA damage of Korean plant food groups at 50 µg/ml concentration according to the GSTM1 genotype. ∗p < 0.05, t-test. Total: mixture of 10 Korean plant food groups.

Fig. 2.

Relative score of lymphocyte DNA damage of Korean plant food groups at 50 µg/ml concentration according to the GSTT1 genotype. ∗ p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, t-test. Total: mixture of 10 Korean plant food groups.

Fig. 3.

Relative score of lymphocyte DNA damage of Korean diet at 250 µg/ml concentration according to the GSTM1 and GSTT1 genotype. ∗p < 0.05, ∗∗∗p < 0.001. NS: not significant, t-test

Fig. 4.

The effect of Korean diet pre-treatment on H₂O₂ -induced DNA damage in human lymphocytes, classified according to GST genotype. Different letters are significantly different among GST genotypes by Duncan's multiple range test.

Table 1.

Intake of plant foods and oils in the Korean diet (KNHANES V-2, 2011)

Food group	Intake²⁾	Food group	Intake
(plant foods and oils)¹⁾	(g/day)	(plant foods and oils)	(g/day)
Cereals		Kimchi
Rice	180.86	Chinese cabbage kimchi	62.98
Rice cake	15.14	Cubed radish kimchi	8.54
Noodles	9.58	Young leafy radish kimchi	5.98
Glutinous rice	10.17	Dong chi mi	5.42
Barley	6.8	Small radish kimchi	6.02
Brown rice	6.6	Na bak kimchi	5.7
Corn	5.53	Watery young leafy radish kimchi	2.95
Flour	5.02	Cucumber kimchi	1.64
Buckwheat noddles	3.73	Beak kimchi	1.24
Noodle soup	3.41	Green onions kimchi	1.1
Total	246.84	Total	101.57
Potatoes		Mushroom
Potato	20.9	Oyster mushrooms	1.33
Sweet potato	13.03	Pine mushrooms	1.36
Chinese noodle	1.21	Oak mushrooms	1.25
Starch	0.71	Winter mushrooms	0.52
Arrow roots	0.58	White mushrooms	0.49
Total	36.43	Phellinus linteus	0.23
Fruits		Total	5.18
Apples	31.12	Legumes
Citrus fruits	34.73	Tofu	21.26
Pears	16.56	Soybean milk	10.91
Water melons	16.49	Soybean	3.8
Grapes	12.63	Kidney beans	0.66
Oranges	8.20	Red beans	0.6
Persimmons	18.94	Total	37.23
Musk melons	8.13	Oils
Banana	6.14	Soybean oil	3.49
Strawberries	5.21	Sesame seed oil	1.47
Peaches	4.87	Wild sesame seed oil	0.21
Japanese apricots	4.61	Rape seed oil	0.27
Kiwies	2.16	Total	5.44
Total	169.79	Nuts
Vegetables		Chestnuts	0.8
Onions	23.68	Sesame	0.65
Red peppers	21.81	Peanuts	0.92
Tomatoes	20.33	Perilla seeds	0.36
Cucumbers	14.93	Walnuts	0.24
Green onions	11.01	Ginko nuts	0.08
Chinese cabbage	10.4	Pine nuts	0.06
Squash	9.78	Total	3.11
Radish leaves	9.14	Seaweeds
Soybean sprouts	8.83	Sea mustard	7.06
Radishes	7.06	Sea tangle	1.91
Spinach	6.64	Laver	1.38
Lettuce	6.57	Green laver	0.34
Carrots	5.43	Total	10.69
Sesame leaf	4.7
Garlic	4.06
Cabbage	3.45
Leeks	2.67
Eggplants	2.45
Bracken	2.44
Chwinamuls	2.18
Total	177.56

¹⁾ Individual items that comprised more than 1% of the total dietary intake by the group selected in this study are representative of the plant foods commonly consumed in the Korean diet.

²⁾ g of edible portion/day/person

Table 2.

Normal dietary pattern of Korean diet (a week_2,000 kcal/day)¹⁾

	Monday	Tuesday	Wednesday	Thursday	Friday	Saturday	Sunday
	KD²⁾1	KD 2	KD 3	KD 4	KD 5	KD 6	KD 7
Breakfast	Sorghum rice Spinach soup Cutlassfish (boiled down in soy sauce) Bellflower salad Stir-fried Korean-leek	Rice with beans Beef radish soup Boiled chicken Laver Cooked crown daisy	Mixed cereals rice Sea mustard soup Steamed egg Cooked spinach Cooked mungbean sprout	Rice with beans Soybean soup Grilled Spanish mackerel Stir-fried oyster mushrooms Radish salsd	Toasted bread Grilled ham Vegetable stick Milk (200 ml)	Rice with red beans Soybean paste soup with cabbage Grilled Mackerel Cooked squash Cooked mungbean sprout	Rice with beans Radish soup Grilled sole (flatfish) Cooked leaf beet Japchae
Lunch	Rice Dried Pollack soup Cooked soybean Grilled tofu Chinese cabbage kimchi	Rice Yukgaejang Soft tofu Cooked sweet potato stem Green laver salad	Rice with raw fish Soybean paste soup with winter mushrooms Rakkyo	Rice Soybean paste soup with dried radish leaves Bulgogi Lettuce Cucumber salad	Soybean paste soup with chinese cabbage Hotstone pot Bibim-bap Fried eggs Cucumber kimchi	Rice Seolleongtang Cooked dried radish leaves Cubed radish kimchi	Noodle soup with clams Fried fish paste Small radish kimchi
Dinner	Barley rice Soybean paste soup with chinese cabbage Fried spicy pork Leafy vegetables Green onions salad	Brown & glutinous rice Curled mallow soup Grilled yellow croaker Squash pancake Chinese cabbage kimchi	Rice with beans Kimchi stew Cooked cabbage Grilled saury Cooked bokchoy	Mixed cereals rice Soft tofu stew Cooked chwinamul Na bak kimchi	Brown & glutinous rice Pollack stew Cooked spinach Grilled deodeok Dotorimuk (Acorn jelly salad)	Mixed cereals rice Soybean paste stew with tofu Pyeonyuk (steamed pork) Fried Bellflower Water parsley salad	Brown & glutinous rice Beef and mushroom stew Stir-fried anchovy Cooked eggplant
Snack	Milk (200 ml) Apple (100 g) Citrus fruit (100 g) Injeolmi (50 g)	Milk (200 ml) Kiwi (100 g) Persimmon (100 g) Jeolpyeon (50 g)	Liquid yogurt (145 ml) Strawberry (200 g) Citrus fruit (100 g) Sweet potato (50 g)	Yoghurt (110 ml) Apple (100 g) Pear (100 g) Potato (130 g)	Citrus fruit (100 g) Banana (100 g)	Milk (200 ml) Orange (200 g)	Milk (200 ml) Strawberry (200 g) Persimmon (100 g)

¹⁾ Reference: 2010 Dietary Reference Intakes for Koreans first revision (The Korean Nutrition Society) 2) KD: Korean diet

Table 3.

Primer sequences used in the PCR reactions

Primer	Sequences (5' ——-3')
GSTM1-forw.	gaa ctc cct gaa aag cta aag c
GSTM1-rev.	gtt ggg ctc aaa tat acg gtg g
GSTT1-forw.	ttc ctt act ggt cct cac act tc
GSTT1-rev.	tca ccg gat cat ggc cag ca
β-Globin-forw.	caa ctt cat cca cgt tca cc
β-Globin-rev.	gaa gag cca agg aca ggt ac

Table 4.

The effect of Korean plant food group pre-treatment on H₂O₂ -induced DNA damages in human lymphocytes, classified according to GST genotype¹⁾

Food Groups	GST genotypes
Food Groups	Group 1 (N = 4) GSTM1+/ GSTT1+	Group 2 (N = 5) GSTM1+/ GSTT1-	Group 3 (N = 4) GSTM1- /GSTT1+	Group 4 (N = 4) GSTM1-/ GSTT1-
Positive control (H₂O₂)²⁾	100	100	100	100
DNA in tail (%)
Potatoes	41.8 ± 3.0^ab3)	39.7 ± 1.7^ab	38.1 ± 2.1^a	45.9 ± 2.0^b
Nuts	32.3 ± 1.3^a	34.2 ± 1.9^a	30.3 ± 1.1^a	39.4 ± 1.2^b
Cereals	47.3 ± 2.7^NS4)	45.7 ± 1.5	43.6 ± 2.2	46.6 ± 1.7
Fruits	17.9 ± 1.0^a	24.6 ± 1.9^b	20.5 ± 1.4^ab	30.0 ± 1.4^c
Kimchi	25.7 ± 1.2^a	32.1 ± 1.4^b	25.9 ± 1.7^a	34.6 ± 1.2^b
Legumes	38.4 ± 1.8^a	38.5 ± 1.4^a	36.0 ± 2.2^a	45.4 ± 1.2^ab
Mushroom	14.4 ± 1.1^a	20.8 ± 1.5^b	13.0 ± 1.0^a	24.8 ± 1.0^c
Oils	50.8 ± 3.3^NS	50.5 ± 2.5	45.9 ± 2.8	48.4 ± 1.9
Vegetables	15.8 ± 0.4^a	23.8 ± 1.6^b	18.8 ± 1.2^a	28.5 ± 1.1^c
Seaweeds	23.5 ± 1.2^a	27.2 ± 1.4^ab	25.5 ± 2.0^ab	29.4 ± 1.6^b
Total⁵⁾	36.9 ± 1.3^a	33.6 ± 2.4^a	31.3 ± 2.1^a	43.3 ± 1.1^b
Tail moment (%)
Potatoes	24.7 ± 1.9^NS	23.8 ± 1.2³⁾	20.8 ± 2.0	23.0 ± 0.8
Nuts	19.2 ± 1.0^bc	18.1 ± 0.8^ab	15.6 ± 1.4^a	21.5 ± 0.4^c
Cereals	30.5 ± 2.4^b	26.3 ± 1.4^ab	23.0 ± 2.4^a	25.6 ± 0.6^ab
Fruits	11.1 ± 0.5^a	12.7 ± 0.5^a	11.2 ± 1.0^ab	14.7 ± 0.4^b
Kimchi	16.4 ± 0.9^ab	17.5 ± 0.8^ab	15.0 ± 1.2^a	19.0 ± 1.4^b
Legumes	23.4 ± 1.7^b	21.9 ± 1.0^ab	19.0 ± 1.7^a	23.1 ± 0.8^b
Mushroom	8.3 ± 0.3^b	9.6 ± 0.4^b	6.3 ± 0.7^a	11.8 ± 0.3^c
Oils	28.8 ± 2.3^ab	30.3 ± 1.4^b	23.4 ± 3.2^a	26.71.1^ab
Vegetables	10.1 ± 0.5^a	12.4 ± 0.4^b	9.0 ± 0.6^a	13.0 ± 0.3^b
Seaweeds	13.7 ± 0.8^a	15.1 ± 0.5^ab	13.7 ± 1.1^a	16.3 ± 0.4^b
Total	20.7 ± 1.3^ab	19.3 ± 0.9^ab	17.7 ± 1.6^a	22.5 ± 0.6^b
Tail length (%)
Potatoes	35.0 ± 1.0^b	28.1 ± 1.4^a	29.4 ± 2.4^a	29.7 ± 1.4^a
Nuts	29.6 ± 1.5^b	23.0 ± 1.5^a	22.7 ± 2.1^a	29.6 ± 0.6^b
Cereals	40.7 ± 2.1^b	30.1 ± 1.9^a	30.7 ± 2.7^a	33.7 ± 0.9^a
Fruits	22.9 ± 1.3^b	18.7 ± 0.8^a	17.9 ± 1.0^a	20.9 ± 1.5^ab
Kimchi	29.9 ± 0.7^c	23.2 ± 1.3^a	25.0 ± 1.9^ab	28.2 ± 1.2^bc
Legumes	34.3 ± 1.0^b	27.4 ± 1.2^a	26.6 ± 1.7^a	29.9 ± 1.4^a
Mushroom	15.4 ± 1.2^a	13.8 ± 0.4^a	12.4 ± 1.4^a	19.0 ± 1.2^b
Oils	36.1 ± 1.3^ab	35.2 ± 1.6^ab	30.3 ± 3.5^a	37.3 ± 0.9^b
Vegetables	21.7 ± 1.4^b	17.6 ± 1.2^a	15.9 ± 1.2^a	21.2 ± 1.2^b
Seaweeds	23.9 ± 1.2^ab	21.2 ± 0.8^a	21.0 ± 1.5^a	25.0 ± 1.1^b
Total	28.1 ± 1.5^ab	24.8 ± 1.0^a	26.2 ± 1.9^ab	29.6 ± 1.4^b

¹⁾ All values are Relative score (%) for positive control (means ± SE)

²⁾ Positive control (H₂O₂): The maximum amount of DNA damage

³⁾ Different letters are significantly different among GST genotypes by Duncan's multiple range test.

⁴⁾ NS: not significant 5) Total: mixture of 10 Korean plant food groups

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