Thymol Reduces Hepatorenal Oxidative Stress, Inflammation and Caspase-3#xd; Activation in Rats Exposed to Indomethacin

Figures & data

Table 1. Body weight gain and relative organ weights of Control, IND alone, IND + THY and THY alone rats for 21 days

	Control	IND alone	IND + THY	THY alone
Body weight gain (g)	20.15 ± 2.04	10.64 ± 1.28^a	15.74 ± 1.38^abc	19.94 ± 2.35^b
Kidney weight (g)	2.03 ± 0.16	2.92 ± 0.04^a	2.68 ± 0.24^abc	2.08 ± 0.12^b
Liver weight (g)	4. 65 ± 0.21	6.45 ± 0.21^a	5.83 ± 0.37^abc	4.58 ± 0.64^b

IND: 5 mg/kg indomethacin, THY: 250 mg/kg thymol. n = 8. Values are expressed as mean ± SEM of 8 rats. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus THY alone.

Table 2. Hepatic function biomarkers in rats treated with IND alone, IND with THY and THY alone for 21 days

Group	Control	IND alone	IND + THY	THY alone
ALB (g/dL)	22.35 ± 2.28	14.47 ± 1.68^a	19.34 ± 2.79^abc	21.78 ± 3.34^b
ALT (U/L)	12.15 ± 1.04	76.16 ± 4.08^a	56.24 ± 2.66^abc	11.67 ± 1.66^b
ALP (U/L)	64.28 ± 3.14	98.27 ± 5.15^a	74.32 ± 4.18^abc	63.45 ± 3.54^b
AST (U/L)	22.46 ± 2.08	58.37 ± 4.02^a	42.18 ± 3.45^abc	21.67 ± 3.07^b
GGT (U/L)	43.28 ± 4.25	84.26 ± 6.36^a	64.12 ± 4.02^abc	42.62 ± 5.12^b
G6PDH (U/L)	0.94 ± 0.02	3.14 ± 0.16^a	2.36 ± 0.12^abc	0.86 ± 0.05^b
GLDH (U/L)	36. 20 ± 4.12	86.12 ± 5.64^a	63.74 ± 4.16^abc	37.04 ± 4.08^b
LDH (U/L)	8.36 ± 1.24	26.46 ± 2.38^a	19.28 ± 1.34^abc	7.88 ± 1.64^b
MDH (U/L)	48.26 ± 3.12	92.22 ± 9.34^a	84.14 ± 7.18^abc	47.34 ± 3.02^b
SDH (U/L)	13.48 ± 2.16	56.24 ± 4.33^a	41.23 ± 4.14^abc	11.89 ± 3.65^b
dBIL (mg/dL)	0.33 ± 0.02	0.69 ± 0.02^a	0.36 ± 0.02^abc	0.32 ± 0.01^b
tBIL (mg/dL)	0.98 ± 0.01	1.04 ± 0.02^a	0.90 ± 0.02^abc	0.96 ± 0.01^b
PNP (U/mg protein)	5.88 ± 0.47	10.32 ± 1.05^a	7.45 ± 0.78^abc	6.01 ± 0.02^b
PON 1 (U/L)	95.21 ± 2.24	53.32 ± 1.14^a	90.23 ± 2.27^abc	94.76 ± 2.16^b
ARG (µmol/mg protein)	12.31 ± 1.47	32.14 ± 2.56^a	16.45 ± 1.34^abc	11.78 ± 1.65^b
CIT (µmol/mg protein)	1.76 ± 0.24	3.14 ± 0.67^a	2.38 ± 0.14^abc	1.65 ± 0.34^b
HYP (µmol/mg protein)	0.98 ± 0.03	2.67 ± 0.21^a	1.67 ± 0.32^abc	1.02 ± 0.04^b

IND: 5 mg/kg indomethacin, THY: 250 mg/kg thymol. n = 8. Values are expressed as mean ± SEM of 8 rats. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus THY alone.

Table 3. Renal function biomarkers in rats treated with IND alone, IND with THY and THY alone for 21 days

Group	Control	IND alone	IND + THY	THY alone
Urea (mg/dL)	25.34 ± 2.57	65.34 ± 4.96^a	53.46 ± 4.12^abc	24.93 ± 2.16^b
Creatinine (mg/dL)	2.08 ± 0.06	4.78 ± 0.35^a	3.02 ± 0.04^abc	1.98 ± 0.47^b
Uric acid (mg/dL)	6.15 ± 0.78	17.02 ± 3.15^a	9.24 ± 0.87^abc	6.50 ± 0.02^b
Cystatin-C (mg/dL)	0.32 ± 0.01	0.74 ± 0.04^a	0.36 ± 0.02^abc	0.30 ± 0.01^b
Na^+ (mmo//L)	84.06 ± 3.12	138.08 ± 12.06^a	96.14 ± 9.84^abc	83.78 ± 3.64^b
K^+ (mmo//L)	7.34 ± 0.13	23.16 ± 5.24^a	14.24 ± 2.18^abc	5.69 ± 1.78^b
HCO3^−(mmo//L)	14.18 ± 2.04	38.67 ± 3.29^a	24.35 ± 2.17^abc	14.26 ± 1.98^b
Cl^− (mmo//L)	33.45 ± 2.42	63.74 ± 4.27^a	46.21 ± 3.18^abc	31.94 ± 3.68^b

IND: 5 mg/kg indomethacin, THY: 250 mg/kg thymol. n = 8. Values are expressed as mean ± SEM of 8 rats. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus THY alone.

Table 4. Serum lipid profile of rats treated with IND alone, IND with THY and THY alone for 21 days

Group	Control	IND alone	IND + THY	THY alone
CHOL (mg/dL)	1.15 ± 0.04	2.76 ± 0.16^a	1.94 ± 0.06^abc	1.12 ± 0.04^b
TRIGS (mg/dL)	1.24 ± 0.02	3.20 ± 0.12^a	1.76 ± 0.12^abc	1.26 ± 0.04^b
HDL (mg/dL)	1.34 ± 0.10	0.65 ± 0.04^a	0.94 ± 0.04^abc	1.30 ± 0.12^b
LDL (mg/dL)	1.16 ± 0.01	2.22 ± 0.18^a	2.14 ± 0.10^abc	1.12 ± 0.02^b

IND: 5 mg/kg indomethacin, THY: 250 mg/kg thymol. n = 8. Values are expressed as mean ± SEM of 8 rats. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus THY alone.

Table 5. Hepatic and renal antioxidant status in rats treated with IND alone, IND with THY and THY alone for 21 days

Group		Control	IND alone	IND + THY	THY alone
SOD	Liver Kidney	23.25 ± 1.21 12.03 ± 0.16	12.10 ± 0.98^a 5.08 ± 0.09^a	16.98 ± 0.67^abc 8.67 ± 0.24^abc	22.95 ± 2.14^b 11.92 ± 0.17^b
CAT	Liver Kidney	15.05 ± 0.91 14.05 ± 0.24	4.13 ± 0.08^a 6.08 ± 0.49^a	9.68 ± 0.10^ab 10.37 ± 0.09^abc	9.43 ± 0.12^ab 14.21 ± 067^b
GPx	Liver Kidney	8.02 ± 0.24 5.02 ± 0.06	2.48 ± 0.09^a 2.01 ± 0.05^a	6.78 ± 0.08^abc 3.47 ± 0.04^abc	6.34 ± 0.07^ab 4.87 ± 0.17^b
GST	Liver Kidney	7.04 ± 0.22 9.17 ± 0.32	3.08 ± 0.06^a 4.28 ± 0.15^a	5.38 ± 0.05^abc 7.21 ± 0.13^abc	6.89 ± 0.04^b 8.95 ± 0.58^b
GSH	Liver Kidney	4.03 ± 0.18 4.16 ± 0.05	1.98 ± 0.02^a 1.88 ± 0.04^a	2.97 ± 0.05^abc 2.67 ± 0.05^abc	3.94 ± 0.27^b 3.91 ± 0.14^b
MDA	Liver Kidney	3.04 ± 0.02 2.48 ± 0.05	8.29 ± 0.16^a 5.35 ± 0.14^a	5.21 ± 0.07^abc 3.56 ± 0.06^abc	3.21 ± 0.04^b 2.35 ± 0.09^b

SOD activity (units/mg protein); CAT activity (μmole H₂O₂ consumed/min/mg protein). GPx activity (μmole GSH/min/mg protein), GST activity (μmole CDNB–GSH complex formed/minute/mg protein). GSH (μg/mg protein), MDA (μmole/mg protein), IND: 5 mg/kg indomethacin; THY: 250 mg/kg thymol. n = 8. Values are mean ± SEM of 8 rats. ^ap < 0.05 versus Control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus Mn alone.

Figure 1. Effect of thymol on hepatic and renal TAC and TOS levels in indomethacin-treated rats. n = 8. Each bar represents mean ± SEM of 8 rats. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone. IND: 5 mg/kg indomethacin; THY: 250 mg/kg thymol.

Figure 2. Influence of thymol on hepatic and renal MPO activity and NO level in indomethacin-treated rats. n = 8.Each bar represents mean ± SEM of 8 rats. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus THY alone. IND: 5 mg/kg indomethacin; THY: 250 mg/kg thymol.

Figure 3. Effect of thymol on hepatic and renal levels of TNF-α, IL-1β and caspase-3 activity in indomethacin-treated rats. n = 8. Each bar represents mean ± SEM of 8 rats in triplicates. ^ap < 0.05 versus control; ^bp < 0.05 versus IND alone; ^cp < 0.05 versus THY alone. IND: 5 mg/kg indomethacin; THY: 250 mg/kg thymol.

Figure 4. Photomicrograph of representatives liver and kidney of (A) Control, (B) IND alone, (C) IND + THY and (D) THY alone. Control liver and kidney showed intact architectural arrangement. Liver of IND alone rats showed infiltration of inflammatory cells, dispution of hepatocytes arrangement and enlargement of the sinusoids; Liver of rat co-treated with IND + THY showing few inflammatory cells while THY alone treated group showing near to normal architecture. The kidney of IND alone showed vacuolation and renal tubular necrosis, disintegration of medullary tubules with edema (arrow) at interstitium and inflammatory cellular infiltration. The kidney of rats co-treated with IND + THY showed mild tubular degeneration and infiltration of inflammatory cells whereas THY alone treated group showing near to normal architecture (H & E X 400).