Antioxidant and hepatoprotective potential of Aegle marmelos Correa. against CCl₄-induced oxidative stress and early tumor events

Figures & data

The treatment schedule is presented below.

Treatment Regimen
Groups	Treatment on 1^st to 6^th day	Treatment on 7^th Day
1. Control	Water 10 mL/Kg Orally	Corn oil 1 mL/Kg b. wt. Orally
2. Only D2	D2 (mg/Kg b. wt.) in 10 mL water, Orally	D2 (mg/Kg b. wt.) in 10 mL water, Orally
3. D1 + Toxicant	D1 (mg/Kg b. wt.) in 10 mL water, Orally	D1 in 10 mL water + CCl4 1 mL/Kg (1:1 in corn oil) Orally
4. D2 + Toxicant	D2 (mg/Kg b. wt.) in 10 mL water, Orally	D2 in 10 mL water + CCl4 1 mL/Kg (1:1 in corn oil) Orally
5.Only Toxicant	Water 10 mL/Kg Orally	CCl4, 1 mL/Kg b. wt. (1:1 in corn oil) Orally

D1 = A. marmelos 25 mg/Kg (methanolic extract was dissolved in water), D2 = A. marmelos 50 mg/Kg (methanolic extract was dissolved in water).

Table I.  Effect of pretreatment of A. marmelos on the CCl₄-mediated depletion in the activity of glutathione content and its metabolizing enzymes in the liver of Wistar rats.

Treatment Groups	Reduced glutathione (n moles GSH/gm tissue)	Glutathione reductase (n moles NADPH oxidized/min/mg protein)	Glutathione-S-transferase (nmol CDNB conjugate formed/min/mg protein)
Group I
Normal pellet diet	0.856 ± 0.014	483.77 ± 25.94	1269.47 ± 29.44
Group II
Only A. marmelos 50 mg/Kg b. wt.	0.789 ± 0.006	451.08 ± 33.96	1252.23 ± 51.04
Group III
A. marmelos 25 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	0.502 ± 0.016^c	396.52 ± 20.56^c	752.23 ± 47.28^b
Group IV
A. marmelos 50 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	0.644 ± 0.002^b	366.95 ± 10.44^c	895.60 ± 24.56^b
Group V
Only CCl4 (1 mL/Kg b wt)	0.419 ± 0.022^a	294.60 ± 15.37^a	525.56 ± 31.63^a

D1 = A. marmelos 25 mg/Kg; D2 = A. marmelos 50 mg/Kg; ^ap ≤ 0.001 on comparison of normal control with CCl₄ treated group and only A. marmelos group; ^bp ≤ 0.001 and; ^cp ≤ 0.005 on comparison of A. marmelos treated groups with only CCl₄ treated group.

Table II.  Effect of pretreatment of A. marmelos on the CCl₄-mediated depletion in the activity of antioxidant enzymes in the liver of Wistar rats.

Treatment Groups	Catalase (n moles H2O2 consumed/min/mg protein)	Glutathione peroxidase (n moles NADPH oxidized/min/mg protein)	Glucose-6-phosphate Dehydrogenase (nmoles NADP reduced/min/mg protein)
Group I
Normal pellet diet	105.79 ± 6.29	428.14 ± 6.48	93.87 ± 13.54
Group II
Only A. marmelos 50 mg/Kg b. wt.	99.91 ± 2.54	433.86 ± 19.9	91.62 ± 12.46
Group III
A. marmelos 25 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	45.67 ± 2.7^a	274.54 ± 31.48^a	62.00 ± 5.71^c
Group IV
A. marmelos 50 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	78.32 ± 3.28^b	360.43 ± 72.62^b	88.92 ± 11.7^b
Group V
Only CCl4 (1 mL/Kg b wt)	12.21 ± 1.39	208.80 ± 8.98^b	29.54 ± 3.61^a

D1 = A. marmelos 25 mg/Kg; D2 = A. marmelos 50 mg/Kg; ^ap ≤ 0.001 on comparison of normal control with CCl₄ treated group and only A. marmelos group; ^bp ≤ 0.001 and; ^cp ≤ 0.005 on comparison of A. marmelos treated groups with only CCl₄ treated group.

Table III.  Effect of pretreatment of A. marmelos on the CCl₄-mediated alteration in the activities of xanthine oxidase, quinine reductase and enhancement of lipid peroxidation in the liver of Wistar rats.

Treatment Groups	Xanthine Oxidase (μg uric acid formed/min/mg protein)	Lipid Peroxidation (n moles MDA formed/hr/gm tissue)	Quinone Reductase (nmoles dichloroindophenol reduced/min/mg protein)
Group I
Normal pellet diet	0.509 ± 0.049	4.15 ± 0.05	587.66 ± 17.7
Group II
Only A. marmelos 50 mg/Kg b. wt.	0.440 ± 0.005	3.91 ± 0.22	532.45 ± 21.63
Group III
A. marmelos 25 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	0.58 ± 0.023^b	7.45 ± 0.33^c	395.74 ± 45.84^b
Group IV
A. marmelos 50 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	0.51 ± 0.028^b	6.56 ± 0.047^b	495.78 ± 28.60^b
Group V
Only CCl4 (1 mL/Kg b wt)	0.917 ± 0.33^a	9.99 ± 0.36^a	137.70 ± 16.53^a

D1 = A. marmelos 25 mg/Kg; D2 = A. marmelos 50 mg/Kg; ^ap ≤ 0.001 on comparison of normal control with CCl₄ treated group and only A. marmelos group; ^bp ≤ 0.001 on comparison of A. marmelos treated groups with only CCl₄ treated group;cp < 0.005.

Table IV.  Effect of pretreatment of A. marmelos on the CCl₄-mediated enhancement of liver toxicity markers in Wistar rats.

Treatment Groups	ALT (IU/L)	AST (IU/L)	LDH (nmol NADH oxidized/min/mg protein)
Group I
Normal pellet diet	24.23 ± 0.59	16.96 ± 0.28	392.66 ± 5.72
Group II
A. marmelos 50 mg/Kg b. wt.	22.07 ± 1.26	16.67 ± 1.42	367.38 ± 8.18
Group III
A. marmelos 25 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	46.71 ± 0.87^b	50.78 ± 6.59^b	626.79 ± 6.14^b
Group IV
A. marmelos 50 mg/Kg b. wt. + CCl4 (1 mL/Kg b wt)	31.19 ± 1.21^b	36.15 ± 3.49^b	533.49 ± 8.35^b
Group V
Only CCl4 (1 mL/Kg b wt)	74.88 ± 4.39^a	94.28 ± 9.17^a	997.68 ± 11.05^a

D1 = A. marmelos 25 mg/Kg; D2 = A. marmeloss 50 mg/Kg; ^ap ≤ 0.001 on comparison of normal control with CCl₄ treated group and only A. marmelos group; ^bp ≤ 0.001 on comparison of A. marmelos treated groups with only CCl₄ treated group.

Figure 1.  Effect of pre-treatment of A. marmelos on CCl₄-induced cell proliferation response. ^bp < 0.001, compared with the corresponding value for treatment with only CCl₄ treated group. ^ap < 0.001 compared to corresponding value for saline treated control group. D1 = A. marmelos 25 mg/Kg; D2 = A. marmelos 50 mg/Kg.

Figure 2.  Effect of pre-treatment of A. marmelos on CCl₄-induced ODC activity. ^bp < 0.001, compared with the corresponding value for treatment with only CCl₄ treated group. ^ap < 0.001 compared to corresponding value for saline treated control group. D1 = A. marmelos 25 mg/Kg; D2 = A. marmelos 50 mg/Kg.