Brazilian Bidens pilosa Linné yields fraction containing quercetin-derived flavonoid with free radical scavenger activity and hepatoprotective effects

Figures & data

Table 1. Groups of animals, experimental pretreatments, and challenge

Group	10-day pretreatment	Dose	Challenge on day 11
Normal control group (NC)	Vehicle	0.5 ml	None
Negative control group (NEG)	Vehicle	0.5 ml	CCl4
Positive control group (SIL)	Silymarin	15 mg/kg	CCl4
HCE group	HCE	15 mg/kg	CCl4
f-EtOAc group	f-EtOAc	15 mg/kg	CCl4

Table 2. Activity of Bidens pilosa (hydroethanol crude extract and fractions) or Silymarin (0.5 to 500 µg/ml) on scavenging of 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH) and hydroxyl radicals (^•OH), inhibition of lipid peroxidation in vitro (LPO), and total polyphenol content in the plant samples (TPC)

Sample	DPPH IC50 (µg/ml)	OH IC50 (µg/ml)	LPO IC50 (µg/ml)	TPC (GAE/mg)
HCE	98.0±1.0^a	14.2±3.2^a	35.8±3.9^a	2.1±0.2
f-EtOAc	32.3±6.7^a	4.3±1.5	25.1±4.5	3.6±0.2^b
f-MeOH	102.9±2.4^a	18.5±8.2^a	90.4±3.4^a	1.0±0.1
f-CHCl3	248.0±1.8^a	33.0±6.0^a	166.3±2.6^a	0.8±0.1
Silymarin	13.9±0.7	7.6±0.8	10.1±4.5	–

Note: Hydroethanol crude extract (HCE), ethyl acetate fraction (f-EtOAc), methanol fraction (f-MeOH), and chloroform fraction (f-CHCl₃). Values are expressed as means±SD (n=3). IC₅₀: half maximal inhibitory concentration was obtained by linear regression.

^aDenotes significant differences at P < 0.05 in comparison to the positive control (SIL).

^bIn comparison to HCE, f-MeOH, or f-CHCl₃.

Table 3. Effects on the plasma ferric reducing/antioxidant power (FRAP), hepatic reduced glutathione content (GSH), and hepatic catalase activity (CAT) in mice pretreated with the hydroethanol crude extract (HCE) or the ethyl acetate fraction (f-EtOAc) from Bidens pilosa (15 mg/kg, p.o., 10 days), after CCl₄ on day 11 (0.5 ml/kg, i.p.), and controls

	Biomarkers of oxidative damage and antioxidant defenses
Group	FRAP (mM)	GSH (µmol/g)	CAT (mmol/min.g)
NC	21.7±0.7^a	34.1±3.1^a	163.7±29.6^a
NEG	11.7±0.8	8.1±3.6	517.6±67.9
f-EtOAc	20.7±0.7^a	30.1±0.4^a	366.4±44.9^a,b
HCE	18.9±0.9^a	25.8±1.5^a,b	447.1±33.1^b
Silymarin	24.5±2.0^a	29.8±4.2^a	441.6±44.3^b

Note: Values are expressed as means ± SD (n=6).

^aDenotes significant statistic differences at P < 0.05 in comparison to the negative control group (NEG).

^bIn comparison to the normal control group (NC).

Fig. 1.  CCl₄ led to increased levels of hepatic lipid peroxidation in treated mice from the negative control group (NEG) compared to the normal control group (NC). The ethyl acetate fraction (f-EtOAC), hydroethanol crude extract (HCE), and the positive control Silymarin (SIL) protected livers from pretreated mice against lipid peroxidation (A); animals from the negative control group (NEG) treated by CCl₄ presented increased levels of carbonyl proteins compared to the normal control group (NC). The f-EtOAC, HCE, and the positive control Silymarin (SIL) protected livers from pretreated mice against oxidative damage to proteins (B). All values are expressed as means±SD, n=6. ^aDenotes significant statistic difference compared to NEG (P<0.05).

Fig. 2.  DNA damage index (0–400) in hepatocytes from mice from the normal control group (NC), negative control group (NEG), positive control group pretreated with Silymarin at 15 mg/kg, p.o. (SIL), and from mice pretreated with the extracts from Bidens pilosa, the hydroethanol crude extract (HCE) or fraction ethyl acetate (f-EtOAc) both at 15 mg/kg (p.o) and on the 11th day treated with CCl₄ (0.5 ml/kg, i.pl). All values are expressed as means ± SD, n=6. ^aDenotes significant difference compared to NEG (P<0.05).

Table 4. Effects of the hydroethanol crude extract (HCE) and the ethyl acetate fraction (f-EtOAc) from B. pilosa and Silymarin against the hepatotoxicity induced in mice by CCl₄

	Serum enzymes
Group	AST (U/L)	ALT (U/L)	LDH (U/L)
NC	9.1±1.1^a	29.2±3.5^a	237.2±38.3^a
NEG	68.4±1.6	91.0±1.5	541.4±41.2
f-EtOAc	12.3±1.4^a	33±10^a	272.4±33.7^a
HCE	21.3±0.9^a,b	48.0±2.1^a,b	295.1±29.3^a
Silymarin	24.4±1.4^a,b	53.0±2.5^a,b	289.6±30.5^a

^aDenotes significant statistic differences at P < 0.05 in comparison to the negative control group (NEG).

^bIn comparison to the normal control group (NC).

Note: Values are expressed as means±SD (n=6).

Abbreviations: Alanine aminotransferase (ALT); aspartate aminotransferase (AST); lactate dehydrogenase (LDH).