Garuga pinnata attenuates oxidative stress and liver damage in chemically induced hepatotoxicity in rats

Figures & data

Table 1. Effects of ethanolic extract of Garuga pinnata on blood biochemical parameters by Paracetamol induced hepatotoxicity (SGOT, SGPT, ACP, ALP and creatinine)

Treatment	SGOT (IU/L)	SGPT (IU/L)	ACP (KA)	ALP (KA)	Creatinine (mg/dl)
Normal control	90.7 ± 14.111	43.3 ± 8.819	1.63 ± 0.2522	4.89 ± 0.3933	1.20 ± 0.2309
Paracetamol (3 g/kg p.o.)	405.3 ± 66.826 ###	293.3 ± 17.638 ####	7.25 ± 1.299 ####	15.29 ± 0.3406 ####	5.07 ± 0.8110 ###
Standard (Silymarin 100 mg/kg)	165.3 ± 37.333 **	66.7 ± 20.276 ***	2.41 ± 0.2205 ***	5.09 ± 0.7091 ****	1.6 ± 0.2309 ***
Garuga pinnata 200 mg/kg	266.7 ± 32.441 Ns	176.7 ± 27.285 *	6.00 ± 0.6614 Ns	14.30 ± 0.5203 Ns	2.53 ± 0.4807 **
Garuga pinnata 400 mg/kg	202.7 ± 19.230 *	103.3 ± 17.638 **	3.50 ± 0.2887 **	8.62 ± 0.7091 ***	1.7 ± 0.1333 ***

All the values are expressed in mean ± S.E.M. (n = 06). *p ˂ 0.05, **p ˂ 0.01 and ***p ˂ 0.001 was considered as statistically significant value when compared with the paracetamol treated group and #p ˂ 0.05, ## p ˂ 0.05 and ###p ˂ 0.001 was considered as statistically significant value when compared with the normal control group. ns- Non significant value where p ˃ 0.05.

Table 2. Effects of ethanolic extract of Garuga pinnata on blood biochemical parameters by Thioacetamide induced hepatotoxicity (SGOT, SGPT, ACP, ALP and creatinine)

Treatment	SGOT (IU/L)	SGPT (IU/L)	ACP (KA)	ALP (KA)	Creatinine (mg/dl)
Normal control	74.7±29.695	96.67±8.119	3.58±0.7949	6.467±0.3406	0.67±0.1333
Thioacetamide 50 mg/kg, s.c.	362.7±23.247 ####	356.7±29.059 ####	9.58±0.6009 ###	15.09±0.5203 ####	4.8±1.538 ###
Standard (Silymarin 100 mg/kg)	165.3±37.333 **	190.00±11.547 ***	4.41±0.3632 ***	7.640±0.3406 ****	1.47±0.3528 **
Garuga pinnata 200 mg/kg	266.7±37.333 Ns	263.3±29.059 *	6.58±0.9391 *	11.56±0.1038 **	2.6±0.2667 *
Garuga pinnata 400 mg/kg	28.3±19.230 *	28.3±12.019 **	5.17±0.7265 **	9.017±0.5178 ***	2.1±0.3528 **

All the values are expressed in mean ± S.E.M. (n = 06). *p ˂ 0.05, **p ˂ 0.01 and ***p ˂ 0.001 was considered as statistically significant value when compared with the thioacetamide treated group and #p ˂ 0.05, ## p ˂ 0.05 and ###p ˂ 0.001 was considered as statistically significant value when compared with the normal control group. ns- Non significant value where p ˃ 0.05.

Table 3. Effects of ethanolic extract of Garuga pinnata on blood biochemical parameters by Ethanol induced hepatotoxicity (SGOT, SGPT, ACP, ALP and creatinine)

Treatment	SGOT (IU/L)	SGPT (IU/L)	ACP (KA)	ALP (KA)	Creatinine (mg/dl)
Normal control	48.00±9.238	18.3±18.559	1.83±0.3005	3.13±0.1933	1.7±0.3528
Ethanol (5 g/kg, 20% w/v p.o.)	298.7±50.877 ###	280.00±17.321 ###	7.17±0.5833 ####	14.50±0.5203 ####	5.8±0.5812 ####
Standard (Silymarin 100 mg/kg)	85.3±29.695 **	136.7±8.819 **	2.25±0.3819 ***	5.680±1.093 ****	2.80±0.6110 **
Garuga pinnata 200 mg/kg	176.00±24.440 Ns	190.00±15.275 *	4.50±1.010 *	9.603±1.415 **	3.2±0.2309 **
Garuga pinnata 400 mg/kg	117.3±32.441 **	163.3±14.530 **	3.91±0.3005 **	7.837±0.5203 ***	2.9±0.3528 **

All the values are expressed in mean ± S.E.M. (n = 06). *p ˂ 0.05, **p ˂ 0.01 and ***p ˂ 0.001 was considered as statistically significant value when compared with the ethanol treated group and #p ˂ 0.05, ## p ˂ 0.05 and ###p ˂ 0.001 was considered as statistically significant value when compared with the normal control group. ns- Non significant value where p ˃ 0.05.

Table 4. Effects of ethanol extract of Garuga pinnata on tissue biochemical estimations by Paracetamol induced hepatotoxicity (SOD, CAT, GSH, LPO and Total proteins)

Treatment	SOD U/min/ g protein	CAT U/mg of protein	GSH nmol/ min/mg protein	LPO moles MDA/ mg proteins/ ml	Total proteins mcg/ml
Normal control	285.71±7.140	4.372±0.0496	0.4632±0.0347	30.96±3.491	188.74±7.140
Paracetamol (3 g/kg p.o.)	64.28±12.373 ###	1.319±0.3284 ####	0.1298±0.0160 ####	178.05±9.820 ####	32.76±8.152 ###
Standard (Silymarin 100 mg/kg)	242.85±7.143 **	3.586±0.0797 ****	0.3357±0.0301 ***	81.28 23.041 **	167.15±9.350 ***
Garuga pinnata 200 mg/kg	142.85±18.896 Ns	2.105±0.0904 *	0.1568±0.0282 Ns	130.64±23.044 Ns	116.14±38.735 *
Garuga pinnata 400 mg/kg	228.56±31.137 **	2.732±0.0821 ***	0.2499±0.0265 *	107.41±21.002 *	153.65±7.140 **

All the values are expressed in mean ± S.E.M. (n = 06). *p ˂ 0.05, **p ˂ 0.01 and ***p ˂ 0.001 was considered as statistically significant value when compared with the paracetamol treated group and #p ˂ 0.05, ## p ˂ 0.05 and ###p ˂ 0.001 was considered as statistically significant value when compared with the normal control group. ns- Non significant value where p ˃ 0.05.

Table 5. Effects of ethanol extract of Garuga pinnata on tissue biochemical estimations by Thioacetamide induced hepatotoxicity (SOD, CAT, GSH, LPO and total proteins)

Treatment	SOD U/min/ g protein	CAT U/mg of protein	GSH nmol/ min/mg protein	LPO moles MDA/ mg proteins/ ml	Total proteins mcg/ml
Normal control	287.5±6.250	4.3384±0.0304	0.4191±0.0224	46.44±7.305	161.75±15.024
Thioacetamide 50 mg/kg, s.c.	87.5±6.250 ####	1.502 ± 0.0975 ####	0.0661±0.0042 ####	174.19±19.761 ###	27.10±2.143 ####
Standard (Silymarin 100 mg/kg)	243.8±10.825 ***	2.3333±0.0692 ****	0.2499±0.0112 ***	81.28±15.989 **	81.34±6.741 **
Garuga pinnata 200 mg/kg	181.3±40.984 *	1.8323±0.0788 Ns	0.1666±0.0191 *	116.12±29.081 Ns	63.26±10.703 *
Garuga pinnata 400 mg/kg	224.9±10.825 **	2.0942±0.1123 **	0.1936±0.0429 **	93.86±9.233 *	74.06±6.288 **

All the values are expressed in mean ± S.E.M. (n = 06). *p ˂ 0.05, **p ˂ 0.01 and ***p ˂ 0.001 was considered as statistically significant value when compared with the thioacetamide treated group and #p ˂ 0.05, ## p ˂ 0.05 and ###p ˂ 0.001 was considered as statistically significant value when compared with the normal control group. ns- Non significant value where p ˃ 0.05.

Table 6. Effects of ethanol extract of Garuga pinnata on tissue biochemical estimations by Ethanol induced hepatotoxicity (SOD, CAT, GSH, LPO and total proteins)

Treatment	SOD U/min/ g protein	CAT U/mg of protein	GSH nmol/ min/mg protein	LPO moles MDA/ mg proteins/ ml	Total proteins mcg/ml
Normal control	278.57±12.370	4.247±0.0746	0.3136±0.0289	59.99±5.886	150.96±9.350
Ethanol (5 g/kg, 20% w/v p.o.)	85.71±12.373 ####	1.4220±0.2276 ####	0.0759±0.0088 ####	155.80±23.784 ##	46.53±1.686 ####
Standard (Silymarin 100 mg/kg)	192.85±12.373 ***	2.436±0.0692 ***	0.1960±0.0200 **	66.76±8.380 **	86.19±4.674 ***
Garuga pinnata 200 mg/kg	135.71±7.140 Ns	1.764±0.0594 Ns	0.1347±0.0171 Ns	93.86±18.840 Ns	60.02±3.447 Ns
Garuga pinnata 400 mg/kg	149.99±12.370 *	2.1967±0.1385 **	0.1642±0.0129 *	86.12±11.890 *	77.83±4.345 **

All the values are expressed in mean ± S.E.M. (n = 06). *p ˂ 0.05, **p ˂ 0.01 and ***p ˂ 0.001 was considered as statistically significant value when compared with the Ethanol treated group and #p ˂ 0.05, ## p ˂ 0.05 and ###p ˂ 0.001 was considered as statistically significant value when compared with the normal control group. ns- Non significant value where p ˃ 0.05.

Figure 1. Histopathology of PCM induced hepatotoxicity model; (1A) Normal control group- Showing well defined liver architecture with normal hepatic cells without any damage, (1B) PCM Treated group- showed the degenerative hepatocytes with necrosis causing severe degree of liver damage, (1 C) Standard silymarin (100 mg/kg) treatment group- showed the mild degree of liver damage and restoring hepatocytes, (1D and 1E) Group treated with Garuga pinnata (200 mg/kg and 400 mg/kg) showed the regeneration of liver cells with mild degree of liver damage

Figure 2. Histopathology of TAA induced hepatotoxicity model; (2A) Normal control group- Showing well defined liver architecture with normal hepatic cells with conserved cytoplasm, (2B) TAA Treated group- showed the degenerative hepatocytes with necrosis causing severe degree of liver damage, (2 C) Standard silymarin (100 mg/kg) treatment group- showed the mild degree of liver damage and restoring hepatocytes, (2D and 2E) group treated with Garuga pinnata (200 mg/kg and 400 mg/kg) showed the regeneration of liver cells with mild degree of liver damage

Figure 3. Histopathology of Ethanol induced hepatotoxicity model; (3A) Normal control group- showing well defined liver architecture with normal hepatic cells with conserved cytoplasm, (3B) ethanol treated group- showed the degenerative hepatocytes with necrosis causing severe degree of liver damage, (3 C) standard silymarin (100 mg/kg) treatment group- showed the mild degree of liver damage and restoring hepatocytes, (3D and 3E) group treated with Garuga pinnata (200 mg/kg and 400 mg/kg) showed the regeneration of liver cells with mild degree of liver damage

Figure 4. Characteristic UV peak of isolated compound at 251 nm indicating the compound belonging to flavonoids group

Figure 5. FTIR spectra of isolated compound showing characteristic peak of flavonoids

Table 7. Functional group characterization using δ ppm of ethyl acetate fraction of Garuga pinnata.

Sr. No	δ ppm values	Description of functional group
1	3.24–3.64	(m, 12 H, Sugar moeity)
2	3.79, 3.81	(d, 1 H, 1 H- Rhamnose)
3	1.11, 1.12	(d, 3 H, CH3- Rhamnose)
4	4.52, 4.63	(d, 4 H, H-1 Glu)
5	5.09, 5.11	(d, 1 H, H-6)
6	6.20, 6.21	(d, 1 H, H-8)
7	6.39, 6.40	(d, 1 H, H-5ʹ)
8	7.61–7.67	(m, 1 H, H-2ʹ, H-6ʹ)

Figure 6. NMR spectra of isolated compound obtained from ethyl acetate fraction of Garuga pinnata.

Figure 7. GC-MS spectra of isolated compound obtained from Garuga pinnata.