Hepatoprotective effects of Tagetes lucida root extract in carbon tetrachloride-induced hepatotoxicity in Wistar albino rats through amelioration of oxidative stress

Figures & data

Figure 1. The chemical composition of Tagetes lucida roots extract as analysed by the HPLC. *mAU means a milli-absorbance unit or 0.001 absorbance units that are used to measure absorbance.

Figure 2. Identification of corynan-17-ol, 18,19-didehydro-10-methoxy-, acetate (ester), the major component of Tagetes lucida root extract; IR. *Y axis; T% means transition percentage, X axis; was wavenumber band (cm⁻¹).

Figure 3. Identification of corynan-17-ol, 18,19-didehydro-10-methoxy-, acetate (ester), the major component of Tagetes lucida root extract; MS. m/z means mass/charge.

Figure 4. Identification of corynan-17-ol, 18,19-didehydro-10-methoxy-, acetate (ester), the major component of Tagetes lucida root extract. Corynan-17-ol, 18,19-didehydro-10-methoxy-, acetate (ester) 17-(acetyloxy)-18,19-didehydro-10-methoxycorynan CAS NO: 56053-13-5. Molecular Formula: C22H28N2O3 and Molecular Weight: 368.46932. m/z means mass/charge.

Table 1. The therapeutic and protective effect of Tagetes lucida root extract on total body weight gain (g) and relative weight of liver (%), CYP2E1 activity, and TNF-α content.

Parameters Groups		Total body weight gain	Relative weight of liver	CYP2E1 activity (nm/min/mg protein)	TNF-α content (pg/ mg protein)
Negative group	(ve- control)	33.90 ± 3.69^c	4.42 ± 0.18^c	3.08 ± 0.29^f	15.2 ± 0.55^d
Positive groups	ve + Silymarin	40.48 ± 3.18^b	3.92 ± 0.30^de	2.80 ± 0.31^f	14.1 ± 0.99^d
	ve + T. lucida	45.73 ± 2.77^a	3.62 ± 0.20^e	2.58 ± 0.56^f	14.44 ± 0.89^d
Therapeutic group	CCl4 Control	10.22 ± 2.23^de	4.34 ± 0.20^cd	11.27 ± 0.78^a	38.83 ± 2.30^a
	Silymarin	10.86 ± 1.81^de	3.71 ± 0.28^e	6.43 ± 0.35^d	17.56 ± 1.34^c
	T. lucida	11.54 ± 0.78^de	3.94 ± 0.35^de	6.87 ± 0.49^c	17.97 ± 0.57^c
protective group	CCl4 Control	-7.03 ± 1.21^f	5.25 ± 0.30^a	9.08 ± 0.56^b	30.01 ± 2.34^b
	Silymarin	8.02 ± 2.87^e	4.88 ± 0.25^b	4.55 ± 0.27^e	13.97 ± 1.32^d
	T. lucida	14.88 ± 1.28^d	4.78 ± 0.30^b	4.70 ± 0.53^e	13.89 ± 1.34^d

Data are presented as the means ± S.D of eight replicates. Data analysed by ANOVA one-way, p ≤ 0.05, Value with the same letter has no significant but value with different letter has significant at 0.05.

Table 2. Effect of Tagetes lucida root extract on liver functions of CCl₄-induced hepatotoxicity in rats.

Main groups	Subgroups	TP mg/dl	Alb mg/dl	Glo mg/dl	Alb/Glo %	T. bilirubin mg/dl	AST U/L	ALT U/L	ALP mg/dl
Negative group Positive group	ve^- control	7.60^c ± 0.32	3.47^bc ± 0.16	4.13^cd ± 0.25	0.85^a ± 0.10	0.99^c ± 0.08	75.70^d ± 3.42	66.64^d ± 3.27	143.12^g ± 2.64
	ve^+ Silymarin	7.58^c ± 0.32	3.45^bc ± 0.09	4.13^cd ± 0.20	0.84^a ± 0.04	0.99^c ± 0.07	77.78^d ± 2.84	68.65^d ± 4.00	141.38^g ± 4.46
	ve^+ T. lucida	8.47^ab ± 0.27	4.04^abb ± 0.33	4.43^bc ± 0.18	0.92^a ± 0.11	0.81^c ± 0.03	76.97^d ± 1.91	52.30^e ± 1.97	136.26^h ± 2.48
Therapeutic group	CCl4 Control	7.66^c ± 0.19	3.29^c ± 0.12	4.37^bc ± 0.20	0.75^a ± 0.05	1.19^b ± 0.09	85.96^c ± 1.50	77.47^c ± 1.54	196.10^c ± 5.68
	Silymarin	8.62^a ± 0.29	3.83^ab ± 0.26	4.80^a ± 0.23	0.80^a ± 0.09	0.92^c ± 0.08	79.19^d ± 3.35	50.12^e ± 2.80	181.15^e ± 2.47
	T. lucida	7.80^bc ± 0.24	3.51^bc ± 0.31	4.29^bc ± 0.24	0.82^a ± 0.11	0.85^c ± 0.07	75.24^d ± 3.01	53.86^e ± 3.22	162.22^f ± 4.33
Protective group	CCl4 Control	6.70^d ± 0.18	2.91^d ± 0.33	3.79^d ± 0.25	0.77^a ± 0.13	2.06^a ± 0.18	122.21^a ± 2.25	238.61^a ± 4.62	226.21^a ± 3.62
	Silymarin	8.00^abc ± 0.73	3.61^bc ± 0.04	4.39^bc ± 0.26	0.83^a ± 0.03	1.20^b ± 0.09	112.51^b ± 3.45	213.74^b ± 4.72	207.31^b ± 2.87
	T. lucida	8.30^abc ± 0.58	3.62^bc ± 0.29	4.68^ab ± 0.34	0.78^a ± 0.08	1.18^b ± 0.11	114.80^b ± 3.54	211.44^b ± 5.65	190.25^d ± 3.52

Data are presented as the means  ±  S.D of eight replicates. Data analysed by ANOVA one-way, p ≤ 0.05, Value with the same letter has no significant but value with different letter has significant at 0.05.

Table 3. Effect of Tagetes lucida root extract on oxidative stress biomarkers and antioxidant biomarkers.

		Oxidative stress biomarkers			Antioxidant biomarkers
Parameters Groups		H2O2 (uM/ g tissue)	MDA (nmol/ g tissue)	MDA (nmol/ ml serum)	CAT (U/ mg protein)	SOD U/mg protein	GSH mg/dL	GR U/mg protein	GST U/mg protein	GPx U/mg protein
Negative group (^-ve Control )		3.55 ± 0.27^b	14.93 ± 0.69^d	7.35 ± 0.15^d	5.78 ± 0.47^e	9.50 ± 0.23^d	13.51 ± 1.30^ef	12.37 ± 2.12^c	10.25 ± 1.58^c	7.33 ± 0.47^d
Positive group	+ve Silymarin	3.86 ± 0.36^b	12.04 ± 1.32^e	7.26 ± 0.13^d	5.79 ± 0.32^e	9.67 ± 0.57^d	14.38 ± 0.69^de	13.44 ± 1.00^c	11.25 ± 1.65^c	7.38 ± 0.30^d
	+ve T. lucida extract	3.52 ± 0.31^b	10.64 ± 0.83^f	7.67 ± 0.16 ^cd	6.12 ± 0.39^e	12.96 ± 0.96^a	14.72 ± 1.82^d	13.03 ± 1.51^c	11.04 ± 1.27^c	9.51 ± 0.93^d
Therapeutic group	CCl4 Control	6.65 ± 0.25^a	24.46 ± 1.63^b	10.81 ± 0.48^a	4.45 ± 0.23^f	6.64 ± 0.43^f	5.58 ± 0.64^g	4.86 ± 0.03^d	4.17 ± 0.26^d	3.72 ± 0.33^e
	Therapeutic Silymarin	3.89 ± 0.18^b	14.76 ± 1.16^d	7.97 ± 0.29^c	8.46 ± 0.64^b	10.55 ± 0.47^c	18.58 ± 1.23^a	17.82 ± 1.01^a	15.25 ± 0.87^a	10.38 ± 1.39^a
	Therapeutic extract	3.57 ± 0.27^b	18.06 ± 0.85^c	8.03 ± 0.48^c	10.00 ± 0.56^a	11.45 ± 0.81^b	16.61 ± 0.99^c	15.83 ± 1.53^b	14.17 ± 1.37^b	10.30 ± 0.43^a
protective group	CCl4 protective Control	6.49 ± 0.29^a	33.63 ± 1.68^a	9.22 ± 0.27^b	4.69 ± 0.13^f	7.43 ± 0.59^e	3.37 ± 0.77^h	2.69 ± 0.52^e	2.36 ± 0.45^e	2.18 ± 0.18^f
	protective Silymarin	3.07 ± 0.26^c	15.37 ± 0.85^d	7.6 ± 0.211^cd	6.44 ± 0.34^d	10.56 ± 0.86^c	13.22 ± 1.46^f	12.94 ± 1.17^c	11.37 ± 1.02^c	9.14 ± 0.29^bc
	protective extract	3.06 ± 0.21^c	17.03 ± 0.74^c	7.65 ± 0.33 ^cd	7.47 ± 0.54^c	11.62 ± 0.39^b	17.62 ± 1.74^b	16.99 ± 1.14^a	14.70 ± 0.99^ab	8.67 ± 0.37^c

Data are presented as the means ± S.D of eight replicates. Data analysed by ANOVA one-way, P ≤ 0.05, Value with the same letter has no significant but value with different letter has significant at 0.05. H₂O₂: hydrogen peroxide; MDA: malondialdehyde the lipid peroxidation biomarker; CAT: catalase; SOD: superoxide dismutase; GSH: glutathione reduced; GR: glutathione reductase; GST: glutathione S transferase; GPx: glutathione peroxidase.

Table 4. The effect of Tagetes lucida roots alcoholic extract in lipid profile of liver.

Groups Parameters		T lipid (mg/g liver)	TC (mg/g liver)	TG (mg/g liver)
Negative group (ve- control)		90.10 ± 8.47^d	30.86 ± 0.55^c	49.05 ± 2.06^c
Positive group	ve + Silymarin	83.44 ± 3.58^e	28.36 ± 2.93^d	47.60 ± 2.24^c
	ve+ T. lucida	89.97 ± 5.52^d	26.11 ± 2.60^e	46.80 ± 1.57^c
Therapeutic group	CCl4 Control	105.76 ± 5.72^b	36.97 ± 2.23^b	67.93 ± 3.26^a
	Therapeutic Silymarin	90.23 ± 3.22^d	21.18 ± 1.82^f	42.39 ± 3.64^d
	Therapeutic T. lucida	94.47 ± 5.55^d	18.54 ± 0.78^d	31.10 ± 3.30^e
Protective group	CCl4 Control	153.08 ± 4.09^a	42.84 ± 2.65^a	52.32 ± 4.11^b
	Protective Silymarin	93.79 ± 3.28^d	25.57 ± 1.45^e	32.89 ± 2.90^e
	Protective T. lucida	101.06 ± 5.92^c	24.89 ± 0.87^e	34.16 ± 0.84^e

Data are presented as the means ± S.D of eight replicates. Data analysed by ANOVA one-way, p ≤ 0.05, Value with the same letter has no significant but value with different letter has significant at 0.05.

Figure 5. A photomicrograph of liver tissue sections from A) Liver of the negative control rats showed the normal structure of this tissue. B) Livers of positive control of silymarin showed normal structure but with slightly dilated blood sinusoids. C) Livers of positive control of Tagetes lucida root extract showed quite normal architecture of liver tissue. D) Livers of therapeutic control showed a significant distortion of the general architecture of the tissue, necrotic cells around the central veins (arrow), and vacuolar degeneration of many cells at the periphery of the lobules (arrowhead). E) Livers of silymarin-treated therapeutic rats showed a marked amelioration of liver tissue except for small focal areas of necrotic cells and/or dilated blood sinusoids. F) Livers of Tagetes lucida root extract-treated therapeutic rats showed liver tissue close to normal but with few inflammatory cells around central veins. G) Livers of the protective control rats showed dilatation and congestion of main blood vessels with fibrosis. The parenchyma of liver tissue shows a focal aggregation of necrotic cells. H) Livers of silymarin-treated protective rats showed inflammatory and necrotic cells around dilated central veins (arrowhead). I) Livers of Tagetes lucida root extract-treated protective rats showed mild protective effect as areas of acidophilic degenerated cells were observed (arrowhead). Hx & E × 400.