Hepatoprotective potential of Phyllanthus muellarianus leaf extract: studies on hepatic, oxidative stress and inflammatory biomarkers

Figures & data

Table 1. Phytochemical constituents of aqueous leaf extract of Phyllanthus muellarianus.

Component	Retention time	Area	Concentration (%)
Catechin	1.266	1541.9765	7.84
Gallic acid	2.516	915.5210	4.65
Quercetin-3-O-glucoside	2.750	589.8275	3.00
Phaselic acid	4.450	219.3390	1.12
Isoquercetin	4.700	443.5140	2.25
Phyllanthusin	5.466	236.4010	1.20
Furosin	11.050	9300.2370	47.28
Geraniin	12.166	3623.6145	18.42
Corilagin	13.700	2370.9260	12.05
Nitidine	14.716	428.3130	2.18

Figure 1. High-performance chromatography profile of Phyllanthus muellarianus aqueous leaf extract.

Table 2. Hepatocellular biomarkers in the serum of acetaminophen-treated rats following oral administration of aqueous leaf extract of Phyllanthus muellarianus.

	ALP	ALT	AST	Albumin	Total bilirubin
Control	12.35 ± 2.11	6.68 ± 0.22	3.35 ± 0.07	5.54 ± 0.80	1.42 ± 0.03
Acetaminophen	32.64 ± 0.67*	27.52 ± 1.17*	15.87 ± 0.64*	2.29 ± 0.15	8.82 ± 0.16
400 mg/kg body weight extract	10.60 ± 0.35Δ	6.44 ± 0.42Δ	4.61 ± 0.22Δ	2.02 ± 0.07Δ	0.86 ± 0.06Δ
100 mg/kg body weight extract + Acetaminophen	28.83 ± 0.67*Δ▿♦	22.26 ± 1.19*Δ▿♦	12.42 ± 0.14*Δ▿♦	3.12 ± 0.21*Δ▿♦	6.36 ± 0.45*Δ▿♦
200 mg/kg body weight extract + Acetaminophen	21.26 ± 1.01*Δ▿♦	17.10 ± 0.81*Δ▿♦	7.93 ± 0.68*Δ▿♦	3.74 ± 0.42*Δ▿♦	4.18 ± 0.32*Δ▿♦
400 mg/kg body weight extract + Acetaminophen	17.03 ± 2.25Δ	9.72 ± 1.77Δ	4.92 ± 0.14Δ	4.98 ± 0.69Δ	3.02 ± 0.17Δ
200 mg/kg body weight sylimarin + Acetaminophen	16.72 ± 1.32Δ	10.44 ± 0.28*Δ	4.12 ± 0.16Δ	4.79 ± 0.18Δ	3.34 ± 0.24Δ

Values are mean ± SEM of 10 determinations and are considered statistically significant at p < 0.05. *p < 0.05, vs control; Δp < 0.05, vs acetaminophen; ∇p < 0.05, vs 400 mg/kg body weight extract; ♦p < 0.05, vs acetaminophen +200 kg/kg body weight sylimarin. ALP: alkaline phosphatase; ALT: alanine aminotransferase; AST: aspartate aminotransferase. ALP, ALT and AST are expressed as nmol/min/mg protein. Albumin and total bilirubin are expressed as mg/dL.

Figure 2. Activities of antioxidant enzymes in acetaminophen-treated mice following oral administration of Phyllanthus muellarianus aqueous leaf extract. Values are mean ± SEM of 10 determinations and are statistically significant at p < 0.05. *p < 0.05, vs control; Δp < 0.05, vs acetaminophen; ∇p < 0.05, vs 400 mg/kg b.w extract; ♦p < 0.05, vs acetaminophen +200 mg/kg b.w sylimarin. SOD: Superoxide dismutase; CAT: Catalase; GSH-Px: Glutathione peroxidase; GSH-Red: Glutathione reductase; Glc 6-PD: Glucose 6-phosphate dehydrogenase. Enzyme activities are expressed in nmol/min/mg protein.

Table 3. Biomarkers of oxidative stress in the liver of acetaminophen-treated rats following oral administration of aqueous leaf extract of Phyllanthus muellarianus.

	GSH	CD	LH	MDA	Protein carbonyl	Fragmented DNA (%)
Control	28.66 ± 2.34	49.38 ± 1.37	31.02 ± 1.25	7.86 ± 0.16	2.75 ± 0.13	2.56 ± 0.35
Acetaminophen	8.44 ± 0.22*	82.16 ± 2.72*	56.34 ± 2.42*	29.28 ± 0.34*	15.04 ± 0.52*	80.52 ± 2.72*
400 mg/kg body weight	30.59 ± 1.80Δ	30.52 ± 0.20Δ	27.54 ± 1.31Δ	4.01 ± 0.07Δ	1.78 ± 0.26Δ	3.66 ± 0.12Δ
100 mg/kg body weight + Acetaminophen	11.88 ± 0.66*Δ▿♦	73.07 ± 1.87*Δ▿♦	44.06 ± 2.73*Δ▿♦	22.81 ± 1.20*Δ▿♦	12.23 ± 0.17*Δ▿♦	63.83 ± 3.25*Δ▿♦
200 mg/kg body weight + Acetaminophen	19.64 ± 1.28*Δ▿♦	65.62 ± 2.67*Δ▿♦	37.12 ± 1.25*Δ▿♦	15.05 ± 1.23*Δ▿♦	7.14 ± 0.62*Δ▿♦	40.14 ± 1.28*Δ▿♦
400 mg/kg body weight + Acetaminophen	24.44 ± 2.17Δ▿	50.63 ± 3.14Δ▿	32.52 ± 3.02Δ	9.06 ± 0.77Δ	3.58 ± 0.27Δ▿	15.65 ± 1.22*Δ▿
200 mg/kg body weight sylimarin + Acetaminophen	22.98 ± 1.78*Δ▿	53.37 ± 3.22Δ▿	33.96 ± 2.69Δ▿	10.97 ± 0.43Δ▿	4.38 ± 0.14Δ▿	20.03 ± 1.27*Δ▿

Values are mean ± SEM of 10 determinations and are considered statistically significant at p < 0.05.

* p < 0.05, vs control; Δp < 0.05, vs acetaminophen; ∇p < 0.05, vs 400 mg/kg body weight extract.

♦ p < 0.05, vs acetaminophen +200 kg/kg body weight sylimarin. GSH: Glutathione reduced; CD: Conjugated dienes; LH: Lipid hydroperoxides; MDA: Malondialdehyde. Glutatione reduced, conjugated dienes, lipid hydroperoxides, malondialdehyde and protein carbonyl are expressed as nmol/mg protein.

Figure 3. Levels of TNF-α, IL-6 and IL-8 in the serum of acetaminophen treated mice following oral administration of Phyllanthus muellarianus aqueous leaf extract. Values are mean ± SEM of 10 determinations and are statistically significant at p < 0.05. *p < 0.05, vs control; Δp < 0.05, vs acetaminophen; ∇p < 0.05, vs 400 mg/kg b.w extract; ♦p < 0.05, vs acetaminophen +200 mg/kg b.w sylimarin. TNF-α: Tumor necrosis factor-α; IL-6: Interleukin-6; IL-8: Interleukin-8. TNF-α, IL-6 and IL-8 are expressed in ng/mL.

Figure 4. Histoartictecture of the liver of acetaminophen treated mice following oral administration of Phyllanthus muellarianus aqueous leaf extract. (A) control, (B) acetaminophen (c) 400 mg/kg b.w extract (d) 100 mg/kg b.w extract + acetaminophen (e) 200 mg/kg b.w extract + acetaminophen (f) 400 mg/kg b.w extract + acetaminophen (g) 200 mg/kg b.w sylimarin + acetaminophen.