Honey and quercetin reduce ochratoxin A-induced DNA damage in the liver and the kidney through the modulation of intestinal microflora

Figures & data

Table 1. Total phenols and flavonoid content in Croatian honey.

Total phenols and flavonoids content in honey
Total phenols (mg GAE/kg honey)	Total flavonoids (mg QUE/kg honey)
572.2 ± 2.05	59.85 ± 3.0 mg

Note: GAE = gallic acid equivalent; QUE = quercetin equivalent.

Table 2. Effect of honey, quercetin and OTA on lactic acid bacteria (LAB) growth (log₁₀ CFU/g).

Experimental groups^a	Strains (log10CFU/g)
	Lactobacillus spp. (mean ± SE)	Statistics vs. OTA	Bifidobacterium spp. (mean ± SE)	Statistics vs. OTA
Control	5.07 ± 0.70	P < .05	6.35 ± 0.23	P < .05
OTA	3.12 ± 0.11	–	3.63 ± 0.19	–
Honey + OTA	3.87 ± 0.33^Δ	–	4.89 ± 0.41^Δ	P < .05
Quercetin + OTA	3.50 ± 0.28^▴	–	4.77 ± 0.51^▴	P < .05
Honey	6.70 ± 0.81*	P < .05	9.32 ± 0.20*	P < .05
Quercetin	5.59 ± 0.20	P < .05	6.95 ± 0.36*	P < .05

Note: CFU: colony forming units.

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg^−1.

*Significantly different compared to control (P < .05);

^ΔSignificantly different compared to honey (P < .05);

^▴Significantly different compared to quercetin (P < .05).

Table 3. Effect of honey, quercetin and OTA on bacterial glucuronidase activity in the cecum content.

Experimental groups^a	β-glucuronidases activity (µmol/g)	Statistics (ANOVA) vs. OTA
Control	0.438 ± 0.04	–
Ochratoxin	0.44 ± 0.02	–
Honey + OTA	0.45 ± 0.05	–
Quercetin + OTA	0.346 ± 0.02*	P < .05
Honey	0.438 ± 0.01	–
Quercetin	0.328 ± 0.06*	P < .05

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.

*Significantly different compared to control (P < .05).

Table 4. Comet assay responses in lymphocytes of the mice treated with honey, quercetin and OTA.

Experimental groups^a	Tail length (μm)				Tail intensity (% DNA)				Tail moment
	Mean ± SE	M	25%	75%	Mean ± SE	M	25%	75%	Mean ± SE	M	25%	75%
Control	16.74 ± 0.68	12.82	11.54	16.03	1.07 ± 0.20^♦	0.00	0.00	0.86	0.23 ± 0.07	0.00	0.00	0.14
OTA	17.09 ± 0.61	14.10	12.82	16.67	2.23 ± 0.32*	0.72	0.00	2.35	0.35 ± 0.06	0.10	0.00	0.32
H + OTA	13.21 ± 0,17*^♦	12.82	11.54	14.10	0.57 ± 0.07*^♦	0.16	0.00	0.85	0.07 ± 0. 01*	0.02	0.00	0.10
QU + OTA	15.85 ± 0.42	13.46	12.18	17.31	0.77 ± 0.09^♦▴	0.18	0.00	1.09	0.12 ± 0.01^▴	0.03	0.00	0.14
H	13.72 ± 0.41*^♦	12.18	11.22	14.10	0.71 ± 0.09^♦	0.04	0.00	0.81	0.09 ± 0.01	0.01	0.00	0.10
Q	15.60 ± 0.49	12.82	12.18	16.03	1.61 ± 0.19	0.35	0.00	1.95	0.24 ± 0.03	0.05	0.00	0.26

Note: Mean ± SE: mean values ± standard error; M: median; 25%: 25% percentile; 75%: 75% percentile.

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.

*Significantly different compared to control (P < .05);

^♦Significantly different compared to OTA (P < .05);

^ΔSignificantly different compared to honey (P < .05);

^▴Significantly different compared to quercetin (P < .05).

Table 5. Comet assay responses in liver cells of the mice treated with honey, quercetin and OTA.

Experimental groups^a	Tail length (μm)				Tail intensity (% DNA)				Tail moment
	Mean ± SE	M	25%	75%	Mean ± SE	M	25%	75%	Mean ± SE	M	25%	75%
Control	16.33 ± 0.30^♦	16.02	14.10	18.59	0.92 ± 0.09^♦	0.26	0.0	1.52	0.14 ± 0.01^♦	0.04	0.0	0.23
OTA	24.87 ± 0.45*	25.00	20.51	28.85	10.59 ± 0.61*	8.72	8.72	17.05	1.97 ± 0.12*	1.58	0.53	3.16
H + OTA	14.23 ± 0.15*^♦	14.10	12.82	15.38	0.00 ± 0.00*^♦	0.0	0.0	0.0	0.00 ± 0.00*^♦	0.0	0.0	0.0
Q + OTA	17.05 ± 0.21^♦▴	16.67	14.74	19.23	0.00 ± 0.00*^♦	0.0	0.0	0.0	0.00 ± 0.00*^♦	0.0	0.0	0.0
H	14.08 ± 0.34*^♦	13.46	12.82	16.67	0.97 ± 0.11^♦	0.10	0.0	1.20	0.14 ± 0.01^♦	0.01	0.0	0.19
QU	15.67 ± 0.30*^♦	14.74	12.82	17.31	0.60 ± 0.11*^♦	0.0	0.0	0.0	0.11 ± 0.02^♦	0.0	0.0	0.0

Note: Mean ± SE: mean values ± standard error; M: median; 25%: 25% percentile; 75%: 75% percentile.

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.

*Significantly different compared to control (P < .05);

^♦Significantly different compared to OTA (P < .05);

^ΔSignificantly different compared to honey (P < .05);

^▴Significantly different compared to quercetin (P < .05).

Table 6. Comet assay responses in kidney cells of the mice treated with honey, quercetin and OTA.

Experimental groups^a	Tail length (μm)				Tail intensity (% DNA)				Tail moment
	Mean ± SE	M	25%	75%	Mean ± SE	M	25%	75%	Mean ± SE	M	25%	75%
Control	14.05 ± 0.15^♦	13.46	12.82	15.38	0.01 ± 0.06^♦	0.00	0.00	0.00	0.00 ± 0.00^♦	0.00	0.00	0.00
OTA	23.36 ± 0.53*	22.76	17.31	29.49	9.92 ± 0.057*	8.90	3.35	15.29	1.69 ± 0.10*	1.38	0.44	2.58
H + OTA	22.97 ± 0.62*^♦Δ	19.23	16.03	29.81	8.30 ± 0.60*^♦Δ	4.89	1.97	13.69	1.54 ± 0.13*^Δ	0.65	0.26	2.38
Q + OTA	19.95 ± 0.51*^♦▴	19.55	14.10	23.72	2.83 ± 0.33*^♦▴	0.70	0.01	3.40	0.52 ± 0.06*^♦▴	0.10	0.00	0.59
H	15.77 ± 0.24*^♦	15.38	13.46	17.31	0.43 ± 0.08*^♦	0.00	0.00	0.089	0.07 ± 0.01*^♦	0.00	0.00	0.01
Q	14.34 ± 0.28^♦	13.46	12.18	16.03	1.05 ± 0.13*^♦	0.23	0.00	1.21	0.14 ± 0.02*^♦	0.03	0.00	0.15

Note: Mean ± SE: mean values ± standard error; M: median; 25%: 25% percentile; 75%: 75% percentile.

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.

*Significantly different compared to control (P < .05);

^♦Significantly different compared to OTA (P < .05);

^ΔSignificantly different compared to honey (P < .05);

^▴Significantly different compared to quercetin (P < .05).

Table 7. Effect of honey, quercetin and OTA on hematological parameters in mice.

Experimental groups^a	Hematological parameters (Mean ± SE)
	WBC (×10^9  L^−1)	RBC (×10^12  L^−1)	Hgb (g L^−1)	Hct	MCV (fL)	MCH (pg)	MCHC (g L^−1)	Plt (g L^−1)
Control	7.01 ± 1.06	8.53 ± 0.27	129.00 ± 2.32	0.41 ± 0.01	193.80 ± 1.68	60.52 ± 1.00	1250.00 ± 26.23	1202 ± 171.92
OTA	7.31 ± 4.68	8.10 ± 0.30	127.20 ± 24.35	0.41 ± 0.06	187.72 ± 0.60*	57.92 ± 5.03	1233.2 ± 109.72	1158.00 ± 319.40
H + OTA	10.36 ± 3.38	11.26 ± 2.59	165.92 ± 7.12*	0.54 ± 0.18*	192.72 ± 2.78	59.12 ± 1.61	1225.2 ± 23.86	1266.00 ± 237.32
QU + OTA	6.70 ± 1.12	8.87 ± 0.99	126.40 ± 11.52	0.43 ± 0.04	191.20 ± 3.68	57.20 ± 3.53	1195.2 ± 58.44	1049.20 ± 410.00
H	9.51 ± 3.56	8.68 ± 0.39	133.40 ± 8.55	0.42 ± 0.02	193.00 ± 2.58	61.52 ± 2.25	1274 ± 34.64	1576.00 ± 171.24*
Q	6.33 ± 2.15	10.30 ± 0.35	148.00 ± 5.09*	0.49 ± 0.02*	191.40 ± 3.83	57.52 ± 1.24	1203.2 ± 3.8296	1197.20 ± 277.44

Note: Mean ± SE: mean values ± standard error; RBC = Red blood cells; Hgb = Hemoglobin; Hct = Hematocrit; MCHC = Mean corpuscular hemoglobin concentration; MCH = Mean corpuscular hemoglobin; MCV = Mean corpuscular volume; Plt, platelets; WBC, total white blood cells.

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.

*Significantly different compared to control (*P < .05).

Table 8. Effect of honey, quercetin and OTA on biochemical parameters in mice.

Experimental groups^a	Biochemical parameters (Mean ± SE)
	AP (U L^−1)	AST (U  L^−1)	ALT (U  L^−1)	LDH (U  L^−1)	Total protein (g  L^−1)	Glucose	Urea
						(mmol  L^−1)	(mmol  L^−1)
Control	83.75 ± 8.54	196.25 ± 107.60	65.00 ± 17.32	1567.50 ± 145.50^♦	62.15 ± 3.52	8.58 ± 2.22	7.38 ± 1.80
OTA	100.00 ± 8.45*	230.00 ± 70.35*	82.50 ± 19.58*	2674.00 ± 191.50*	55.00 ± 3.34*	8.79 ± 1.38	9.00 ± 0.70*
H + OTA	75.00 ± 14.72	183.75 ± 49.23	65.00 ± 14.72	1605.00 ± 217.85^♦	57.75 ± 4.73*	7.42 ± 0.81	5.75 ± 2.02
QU + OTA	71.25 ± 7.50	137.50 ± 25.33	51.25 ± 10.31	1584.00 ± 170.50^♦	54.75 ± 6.65*	8.33 ± 1.74	9.50 ± 2.86
H	83.75 ± 16.01	198.75 ± 65.25	60.00 ± 10.80	1221.50 ± 300.20^♦	58.65 ± 3.68	8.68 ± 1.40	7.88 ± 0.48
Q	88.75 ± 13.15	182.50 ± 51.25	51.25 ± 4.79	980.00 ± 220.05*^♦	66.00 ± 4.78	7.13 ± 2.11	8.5 ± 0.71

Note: Mean ± SE: mean values ± standard error; AST, aspartate aminotransferase; ALT, alanine aminotransferase, AP, alkaline phosphatase, blood glucose levels (glucose), lactate dehydrogenase (LDH).

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.

*Significantly different compared to control (P < .05);

^♦Significantly different compared to OTA (P < .05).

Table 9. Cell number and macrophage spreading index of mice treated with honey, quercetin and OTA.

Experimental groups^a	Total cells ×10^6 (Mean ± SE)	Macrophage spreading index (%)
Control	5.78 ± 1.60	37.00 ± 4.98
Ochratoxin	6.99 ± 1.29	52.05 ± 4.99
Honey + ochratoxin	5.31 ± 1.82	39.97 ± 1.30
Quercetin + ochratoxin	6.29 ± 1.46	46.25 ± 2.50
Honey	5.39 ± 2.17	45.31 ± 5.81
Quercetin	5.20 ± 2.14	46.56 ± 4.99

^aMice were treated ig with bee honey, quercetin and OTA alone or in combination every day for 15 days in the following doses: honey, 2 g kg⁻¹; quercetin, 50 mg kg⁻¹; OTA, 100 μg kg⁻¹.