Hepatoprotective and antioxidant activities of Dicranopteris linearis leaf extract against paracetamol-induced liver intoxication in rats

Figures & data

Table 1. Experimental design on hepatoprotective effect of EADL against PCM-induced rats.

Group	Type of treatment	Pre-treatment^a	Dose (mg/kg)	Hepatotoxic inducer^b
1	Normal control	10% DMSO	–	10% DMSO
2	Hepatotoxic control	10% DMSO	–	3 mg/kg PCM
3	Positive control	Silymarin	200 mg/kg	3 mg/kg PCM
4	Treatment	EADL	50 mg/kg	3 mg/kg PCM
5	Treatment		250 mg/kg	3 mg/kg PCM
6	Treatment		500 mg/kg	3 mg/kg PCM

^aAdministered orally and daily for 7 consecutive days.

^bAdministered orally 3 h after the last pre-treatment on 7^th day.

All test solutions were administered in the volume of 10 ml/kg.

Table 2. TPC value, free radical scavenging activity and antioxidant capacity of EADL.

Sample	Concentration (µg/ml)	Standard drug	TPC value (mg GAE/1 g dry weight of extract)^a	Free radical scavenging activity		Antioxidant capacity
				DPPH (% AA)	SOA (% AA)	ORAC^b (mmol TE/1 g dry weight of extract)
EADL	200	Gallic acid	352.2 ± 48.4	–	–	–
		Ascorbic acid	–	93.7 ± 3.0	–	–
			–	–	92.6 ± 2.2	–
		Trolox	–	–	–	555 ± 12.7

All values are expressed as mean ± SEM.

% AA refers to percentage of antioxidant potential.

^aData, expressed as TPC mg GAE/1 g dry weight of extract, were mean values of triplicate wells in duplicate experiments., Standard error of mean (SEM) < 5%.

^bData, expressed as ORAC value mmol Trolox Equivalent (TE)/1 g dry weight of extract, were mean values of triplicate wells in duplicate experiments, standard error mean (SEM) < 30%.

Table 3. Effect of EADL on in vitro inflammatory activity mediated by LOX and XO.

Sample	Concentration (µg/ml)	Inhibition of Inflammation (%)
		LOX activity	XO activity
EADL	100	25.9 ± 4.4	1.5 ± 0.2

All values are expressed as mean ± SEM.

Note: H, high (71–100%); M, moderate (41–70%); L, low (0–40%); NA, not active (Ismail Suhaimy et al. 2017).

Table 4. Effect of EADL pre-treatment on the body weight (BW), liver weight (LW) and their ratio (LW/BW) in PCM intoxicated rats.

Group	Treatment	Dose (mg/kg)	Body weight (BW) (g)	Liver weight (LW) (g)	LW/BW (%)
Normal Control	Control	–	208.7 ± 5.6	5.9 ± 0.3	2.8 ± 0.1
Negative Control	DMSO + PCM*	–	219.5 ± 4.7	9.7 ± 0.9^a	4.4 ± 0.4^a
Positive Control	Silymarin + PCM*	200	209.4 ± 4.7	6.9 ± 0.2^b	3.3 ± 0.2^b
Treatment	EADL + PCM*	50	215.7 ± 6.6	7.1 ± 0.5^b	3.3 ± 0.3^b
		250	219.8 ± 7.1	7.8 ± 0.2^b	3.6 ± 0.4^b
		500	217.5 ± 4.7	6.2 ± 0.4^b	2.9 ± 0.3^b

Values are expressed as means ± S.E.M. of six replicates.

*PCM was administered in the dose of 3 mg/kg.

^aSignificant different as compared to normal control, p < 0.05.

^bSignificant different as compared to negative control, p < 0.05.

Table 5. Effect of EADL pre-treatment on the levels of ALT, AST, ALP, and TB in PCM intoxicated rats.

Group	Treatment	Dose (mg/kg)	ALT (U/L)	AST (U/L)	ALP (U/L)	TB (umol/L)
Normal control	Control	–	15.8 ± 2.9	95.1 ± 5.9	115.7 ± 7.0	0.5 ± 0.2
Negative control	DMSO + PCM*	–	1714.3 ± 142.2^a	2266.2 ± 340.4^a	330.0 ± 42.4^a	4.1 ± 0.8^a
Positive control	Silymarin + PCM*	200	474.5 ± 82.2^b	690.9 ± 146.6^b	195.5 ± 11.1^b	2.3 ± 0.3^b
Treatment	EADL + PCM*	50	1024.4 ± 230.1^b	971.7 ± 226.1^b	377.8 ± 53.7	1.1 ± 0.2^b
		250	608.9 ± 340.0^b	800.8 ± 291.0^b	225.0 ± 10.0^b	1.3 ± 0.4^b
		500	108.7 ± 29.0^b	313.0 ± 66.0^b	198.5 ± 19.8^b	1.5 ± 0.3^b

Values are expressed as means ± S.E.M. of six replicates.

*PCM was administered in the dose of 3 mg/kg.

^aSignificant different as compared to normal control, p < 0.05.

^bSignificant different as compared to negative control, p < 0.05.

Table 6. Effect of EADL pre-treatment on the levels of SOD, CAT and MDA in liver of PCM intoxicated rats.

Group	Treatment	Dose (mg/kg)	CAT (U/g tissue)	SOD (U/g tissue)	MDA (µM)
Normal control	DMSO + DMSO	–	114.8 ± 1.6	9.7 ± 0.4	2.6 ± 0.6
Negative control	DMSO + PCM*	–	92.9 ± 1.9^a	4.0 ± 0.1^a	5.0 ± 0.6^a
Positive control	Silymarin + PCM*	200	109.5 ± 4.6^b	11.8 ± 1.5	2.6 ± 0.3^b
Treatment	EADL + PCM*	50	96.9 ± 2.7	5.3 ± 0.9	2.6 ± 0.7^b
		250	112.7 ± 5.7^b	7.9 ± 0.2^b	3.0 ± 0.3^b
		500	115.2 ± 3.4^b	8.3 ± 0.5^b	2.7 ± 0.7^b

Values are expressed as means ± S.E.M. of six replicates.

*PCM was administered in the dose of 3 mg/kg.

^aSignificant different as compared to normal control, p < 0.05.

^bSignificant different as compared to negative control (DMSO + PCM), p < 0.05.

Figure 1. (a) Normal liver tissue, b) PCM intoxicated liver tissue (negative control) showed large area of haemorrhagic necrosis around centrilobular region, and inflammatory cell infiltration at the centre of the necrotic foci, c) Effect of 200 mg/kg silymarin pre-treatment on PCM intoxicated liver tissue showing preservation of normal hepatocytes, d) Effect of 50 mg/kg EADL pre-treatment on PCM intoxicated liver tissue showing mild sinusoidal congestion and cellular swelling, e) Effect of 250 mg/kg EADL pre-treatment on PCM intoxicated liver tissue showing moderate haemorrhagic necrosis in centrilobular region and presence of inflammatory infiltrate, f) Effect of 500 mg/kg EADL pre-treatment on PCM intoxicated liver tissue showing mild inflammatory infiltrate and mild cellular swelling. (H&E staining, 100x magnification). CV) Central vein. IF) Inflammatory infiltrate. HN) Haemorrhagic necrosis. SC) Sinusoidal congestion.

Table 7. Histopathological scoring of the liver section of PCM intoxicated rats with or without EADL pre-treatment.

Treatment	Dose (mg/kg)	Steatosis	Necrosis	Inflammation	Hemorrhage
Control	−	−	−	−	−
DMSO + PCM		+	+++	++	+
Silymarin + PCM	200	−	+	−	−
EADL + PCM	50	−	+	−	−
	250	+	++	+	−
	500	+	−	−	−

The severity of various features of hepatic injury was evaluated based on those following scoring scheme:

− normal; + mild effect; ++ moderate effect; +++ severe effect.

Table 8. Comparison on the presence of phytochemical constituents in different partitions of MEDL.

Sample	Phytochemical constituents						Conclusion
	Alkaloids	Saponins	Flavonoids	Tannins and Polyphenolic compound	Triterpenes	Steroids
PEDL	−	1+	−	−	−	1+	Saponins and steroids only presence.
EADL	−	3+	1+	2+	−	2+	Saponins, flavonoids, tannins and polyphenolic compound and steroids presence.
AQDL	−	1+	−	−	1+	−	Saponins and triterpenes only presence.

Table 9. Peak with UV-vis spectra that was a characteristic of a flavonoid-based bioactive compounds.

Peak	RT (min)	Band A	Band B	Types of flavonoids^a
1	19.3	347.0	264.9	Flavones
2	19.9	313.6	256.7	Flavones
3	20.8	352.9	255.5	Flavonols
4	22.7	347.0	264.9	Flavones
5	23.1	348.2	255.5	Flavones
6	26.8	342.2	263.8	Flavones

^aAccording to the UV-vis spectra range provided by Tsimogiannis et al. (2007).

Ismail Suhaimy NW, Noor Azmi AK, Mohtarrudin N, Omar MH, Md. Tohid SF, Cheema MS, Teh LK, Salleh MZ, Zakaria ZA. 2017. Semipurified ethyl acetate partition of methanolic extract of Melastoma malabathricum leaves exerts gastroprotective activity partly via its antioxidant-antisecretory-anti-inflammatory action and synergistic action of several flavonoid-based compounds. Oxid Med Cell Longev. 2017:6542631.

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