Hepatoprotective and free radical scavenging actions of quercetin nanoparticles on aflatoxin B1-induced liver damage: in vitro/in vivo studies

Figures & data

Figure 1. (A) TEM image of quercetin nanocrystals. The size of quercetin nanocrystals was in the range of 30–60 nm with the average size 50 nm. (B) Dynamic light scattering (DLS) of the quercetin nanocrystals. (C) Zeta potential of nanoparticles with the average amount of −24 mV.

Figure 2. (A) Effect of the AFB1 on the viability of rat hepatocytes. (B) Effect of quercetin (Q) (100 μM), Q-NPs (quercetin nanoparticles) (100 μM), N-acetyl cysteine (NAC) (200 μM) on the viability of AFB1-treated hepatocytes. Isolated rat hepatocytes were incubated for 4 h with DMEM containing different concentrations of AFB1. Antioxidants were added 1 h after AFB1 (25 μM) treatment. The viability was determined by MTT reduction assay and expressed as percentage of control. Data are expressed as mean ± SEM of three independent experiments. *Different from control group (p < .05). ^aQ (100 μM) and/or NAC (200 μM) significantly decreased hepatocyte membrane lysis compared to AFB1-treated hepatocytes. **Q-NPs significantly decreased hepatocyte membrane lysis compared to AFB1 and Q-treated hepatocytes.

Figure 3. (A) Effect of quercetin (Q) (100 μM), Q-NPs (quercetin nanoparticles) (100 μM), N-acetyl cysteine (NAC) (200 μM) on the ROS generation of AFB1-treated hepatocytes. ROS is measured as pmole DCF formed/mg protein. (B) TBARS formation induced by AFB1 and the protective effects of 100 μM Q (quercetin), N-acetyl cysteine (NAC) (200 μM) and Q-NPs (quercetin nanoparticles). Isolated rat hepatocytes were incubated for 4 h with DMEM containing AFB1 (25 μM). Data are expressed as mean ± SEM of three independent experiments. *Different from control group (p < .05). ^aQ (100 μM) and/or NAC (200 μM) significantly decreased hepatocyte membrane lysis compared to AFB1-treated hepatocytes. **Q-NPs significantly decreased hepatocyte membrane lysis compared to AFB1 and Q-treated hepatocytes.

Figure 4. Effect of quercetin (Q) (100 μM), Q-NPs (quercetin nanoparticles) (100 μM), N-acetyl cysteine (NAC) (200 μM) on the glutathione depletion induced by AFB1 in primary rat hepatocytes. GSH level is measured as nmole/mg protein. Isolated rat hepatocytes were incubated for 4 h with DMEM containing different AFB1 (25 μM). Data are expressed as mean ± SEM of three independent experiments. *Different from control group (p < .05). ^aQ(100 μM) and/or NAC (200 μM) significantly decreased hepatocyte membrane lysis compared to AFB1-treated hepatocytes. **Q-NPs significantly decreased hepatocyte membrane lysis compared to AFB1 and Q-treated hepatocytes.

Table 1. Effect of Q and Q-NPs on liver function in AFB1-admministrated rats.

Treatment	Control	AFB 1	Q (10 mg/kg)+AFB 1	Q-NPs (10 mg/kg)+AFB 1
AST (U/L)	46.17 ± 5.35	89.34 ± 4.93^a	72.33 ± 6.80^b	54.17 ± 6.32^c
ALT (U/L)	58.12 ± 4.78	94.37 ± 6.17^a	81.33 ± 6.70^b	63.71 ± 9.33^c
ALP (U/L)	64.33 ± 5.29	136.50 ± 3.82^a	115.50 ± 4.53^b	92.48 ± 5.61^c

Values were expressed as mean ± SEM for five animals in each observation.

^a Significantly different from control group (p < .05).

^b Significantly different from AFB1-intoxicated group (p < .05).

^c Significant difference in AFB1 groups treated with Q (p < .05).

ALP: alkaline phosphatase; ALT: alanine transaminase; AST: aspartate transaminase; Q: quercetin, Q-NPs; quercetin nanoparticles.

Figure 5. Effect of quercetin (Q) (100 μM), Q-NPs (quercetin nanoparticles) (100 μM), N-acetyl cysteine (NAC) (200 μM) on the mitochondrial membrane potential in AFB1-treated rat hepatocytes. MMP level is measured as nmole Rh 123/mg protein. Isolated rat hepatocytes were incubated for 4 h with DMEM containing AFB1 (25 μM). Data are expressed as mean ± SEM of three independent experiments. *Different from control group (p < .05). ^aQ (100 μM) and/or NAC (200) significantly decreased hepatocyte membrane lysis compared to AFB1-treated hepatocytes. **Q-NPs significantly decreased hepatocyte membrane lysis compared to AFB1 and Q-treated hepatocytes.

Table 2. Effect of the quercetin and its nanoparticles on liver function markers, antioxidant enzyme activities and lipid peroxidation in rats with aflatoxin B1-induced hepatotoxicity.

Treatment	Control	AFB 1	Q (10 mg/kg)+AFB 1	Q-NPs (10 mg/kg) AFB 1
SOD (U/mg protein)	6.13 ± 0.36	1.62 ± 0.79^a	3.48 ± 0.13^b	5.71 ± 0.64^c
CAT (K/mg protein)	57.21 ± 1.55	18.56 ± 1.42^a	31.03 ± 2.76^b	54.67 ± 3.46^c
GPx (U/mg protein)	72.29 ± 2.45	32.07 ± 1.71^a	48.33 ± 2.63^b	68.12 ± 5.70^c
MDA (nmol/mg protein)	34.17 ± 3.61	145.82 ± 6.13^a	92.77 ± 4.29^b	41.48 ± 3.20^c

Values were expressed as mean ± SEM for five animals in each observation.

^a Significantly different from control group (p < .05).

^b Significantly different from AFB1-intoxicated group (p < .05).

^c Significant difference in AFB1 groups treated with Q (p < .05).

Q: quercetin; Q-NPs: quercetin nanoparticles.