. 2021 Aug 11;13(8):561. doi: 10.3390/toxins13080561

Table 4.

Combinatorial interaction between DON and ZEN.

Cell Line Used	Toxin Levels	Exposure Time and Route	Conclusions	Observations	Comment	References
HepaRG	DON 0.2–10 µM ZEN 1.5–75 µM	48 h	Synergistic at 48 h cell viability	Cell viability	Doses correspond to IC50 values after 48 h	[105]
HepaRG	DON 7.35 µM ZEN 55.1 µM	18 h	Additive at 18 h cell mortality	Cell mortality	Doses correspond to IC50 values after 48 h	[105]
HepaRG	DON 2.5 µM ZEN 0.24 µM	14 days 28 days 42 days	Additive	Cell viability	Doses correspond to maximum levels permitted in cereals for humans	[106]
HepaRG	DON 0.2 µM ZEN 20 µM	1 and 24 h	Synergistic at 1 h Antagonistic at 24 h	Cell proteome	Doses correspond to IC10 values after 48 h	[107]
BEL-7402	DON 0.37–16.9 µM ZEN 0.31–31.4 µM	24 h	Additive	Cell viability	Mixtures used DON + ZEN 0.37 + 0.31 µM 0.68 + 0.63 µM 1.69 + 1.57 µM 3.7 + 3.1 µM 6.8 + 15.7 µM 16.9 + 31.4 µM	[108]
HepG2	DON 0.34–67.5 µM ZEN 15.7–157 µM	24 h	Synergistic	Cell viability	Mixtures used DON + ZEN 0.034 + 15.7 µM 1.69 + 47.1 µM 3.7 + 62.8 µM 37 + 126 µM 67.5 + 157 µM	[109]
HepG2	DON 0.02–2 µM ZEN 0.28–34.5 µM	48 h	Synergistic	Cell viability	DON + ZEN 0.02 + 0.28 µM 0.03 + 0.53 µM 0.07 + 1.1 µM 0.14 + 3 µM 0.27 + 4.4 µM 0.51 + 8.6 µM 1 + 17.3 µM 2 + 34.5 µM	[110]
RAW 246.7	DON 0.0027–0.34 µM ZEN 0.28–37.69 µM	48 h	Synergistic	Cell viability	DON + ZEN 0.0027 + 0.28 µM 0.0054 + 0.6 µM 0.01 + 1.19 µM 0.02 + 2.36 µM 0.04 + 4.71 µM 0.08 + 9.42 µM 0.17 + 18.8 µM 0.34 + 37.69 µM	[110]
Caco-2	DON 3.3–16.7 µM ZEN 10–50 µM	24 h	Antagonistic	Cell viability	DON and ZEN combination in 1:3 ratio	[112]
HCT116	DON 100 µM ZEN 40 µM	24 h	Antagonistic	Cytotoxicity, mitochondrial apoptosis	Doses correspond to IC30 values after 24 h	[113]
HCT116	DON 100 µM ZEN 40 µM	48 h	Antagonistic	Cell cycle analysis	Doses correspond to IC30 values after 24 h	[113]
IPEC-J2	Cytotoxic concentration DON 2 µM DON ZEN 40 µM	48 h	Cytotoxic concentration Reported as non-additive	Cell viability	Dose correspond to cytotoxic and non-cytotoxic concentrations	[90]
IPEC-J2	Non-cytotoxic concentration DON 0.5 µM ZEN 10 µM	48 h	Non-cytotoxic concentration Synergistic	Cell viability		[90]
PK15	DON 0.25 µM ZEN 20 µM	24 h	Synergistic	ROS levels Apoptosis	Doses used are concentrations close to IC10 concentration which were 0.157 and 27.583 µM for DON and ZEN, respectively	[114]
Porcine splenic lymphocytes	DON + ZEN 0.2 + 0.25 µM 1 + 1.26 µM 5.1 + 6.28 µM	48 h	Synergistic	Apoptosis Oxidative injury		[115]
Porcine lymphocytes	DON + ZEN 0.24 + 15.7 μM 0.71 + 31.4 μM	24/48/72 h	Antagonistic	Cell viability	Doses used were below IC50 concentration after 24, 48, 72 h exposure	[116]
Porcine lymphocytes	DON + ZEN 0.24 + 15.7 μM 0.71 + 31.4 μM	24/48/72 h	Antagonistic at lower concentration Synergistic at 72 h at higher concentration	Genotoxicity		[116]
THP-1	DON 0.1–10 μM ZEN 2–100 μM	48 h	Antagonistic	Cell viability	DON + ZEN + 2 μM 0.8 + 16 μM 2 + 40 μM 4 + 80 μM 10 + 100 μM	[117]
ANA-1	DON 0–33.7 µM ZEN 0–37.7 µM	24 h	Synergistic	Cell viability and apoptosis	DON + ZEN concentration used for apoptosis and metabolism study 0.34 + 25.1 µM	[118]
ANA-1	DON 0–33.7 µM ZEN 0–37.7 µM	24 h	Antagonistic	Cell metabolism		[118]
BF-2	DON 0–16.2 µM ZEN 0–120.3 µM	48 h	Antagonism	Cell viability fish Oxidative stress fish	DON + ZEN 0.13 + 1.33 0.25 + 2.66 0.51 + 5.32 1.01 + 10.64 2.02 + 21.29 4.05 + 42.58 8.1 + 85.15 16.2 + 170.3	[119]
Caco-2, HepaRG and THP-1 Co-culture	DON + ZEN Concentration used when Caco-2 cells were in luminal compartment: 1.6 + 24 µM 3 + 31 µM Concentration used when HepaRG cells were in luminal compartment: +20 µM 2.3 + 33 µM	48 h	No cytotoxicity with low concentration and in tri-culture Synergistic effect with higher concentration in bi-culture system	Cell viability		[120]
Animal used	Toxin levels	Exposure time and route	Conclusions	Observations	Comment	References
Zebrafish larvae	DON 67.5 µM ZEN 6.28 µM	72 h	Antagonistic	Cell mortality		[119]
Mice	DON 1500, 2500 µg/kg bodyweight ZEN 20,000, 30,000 µg/kg bodyweight	12 days Intraperitoneal injection	Antagonistic	Oxidative stress Renal apoptosis	DON + ZEN 1500 + 20,000 1500 + 30,000 2500 + 20,000 25 + 30,000 µg/kg bodyweight	[121]
Mice	DON 5000 µg/kg bodyweight ZEN 5000 µg/kg bodyweight	2 weeks Oral administration	Antagonistic	Oxidative stress	No change observed on liver weight	[111]
Mice	DON 5000 µg/kg bodyweight ZEN 5000 µg/kg bodyweight	2 weeks Oral administration	Synergistic	Apoptosis	No change observed on liver weight	[111]
Rats	DON 30 µg/animal/day ZEN 15 µg/animal/day	14 days Administered as a gavage dose	Antagonistic	Liver weight Glutathione level in liver Malondialdehyde level in kidney	Doses are according to EU limits in finished feed for young pigs	[122]
Mice	DON 0.5–2 μM ZEN 10–40 μM	24 h	Synergistic	Cell viability Immune function	DON and ZEN combined 1:20	[123]
Rats	DON 16.5 µg/animal/day ZEN 12.75 µg/animal/day	5 days Intraperitoneal administration	Synergistic	Glutathione and glutathione peroxidase activity in the liver	Doses correspond to 1 mg/kg diet for DON and 1.5 mg/kg diet for ZEN which are close to EU limits in finished feed for young pigs	[124]
Mice	DON 2000 mg/kg ZEN 20,000 mg/kg	21 days Intragastric administration	Antagonistic	Metabolic profiling of liver and serum		[125]
Mice	DON 2000 mg/kg ZEN 20,000 mg/kg	3 weeks Intragastric administration	Antagonistic	Metabolic pathway		[126]
Mice	DON 1500, 2500 µg/kg body weight ZEN 20,000, 30,000 µg/kg body weight	4 days Intraperitoneal injection	Synergistic	Apoptosis Antioxidant levels	DON + ZEN 1500 + 20,000 1500 + 30,000 2500 + 20,000 2500 + 30,000	[127]
Female piglets	DON 1000.6 µg/kg ZEN 269.1 µg/kg DON + ZEN 1007.5 + 265.4 µg/kg	3 weeks Ad libitum feeding	Synergistic	Body weight gain Average daily feed intake Intestinal functions	Barley naturally contaminated with DON and corn naturally contaminated with ZEN was used to manufacture the feed	[128]