Pharmacokinetic Interactions of Antihepatotoxic Wedelolactone with Paracetamol in Wistar Albino Rats

Figures & data

Figure 1 Aseptically germinated seedlings of Eclipta alba. (Linn.) Hassk.

Table 1. Effects of plant growth regulators on initiation of callus in MS medium.

Growth regulators	Days of callus initiation	Extent of callus initiation^a.
IAA (5.7 mg/L)	−	−
2,4-D (13.58 mg/L)	10–12	+++
6-BA (4.43 mg/L)	10–12	++
2,4-D (4.52 mg/L) + IAA (5.7 mg/L)	12–15	+
Kin (4.64 mg/L)	12–15	+
2,4-D (13.58 mg/L) + Kin (4.64 mg/L)	12–15	++
6-BA (4.43 mg/L) + Kin (4.64 mg/L)	10–12	+
IAA (5.7 mg/L) + 6-BA (4.43 mg/L)	12–15	+
Kin (4.64 mg/L) + IAA (5.7 mg/L)	−	−
2,4-D (4.52 mg/L) + 6-BA (4.43 mg/L)	5–7	+++
2,4-D (4.52 mg/L) + 6-BA (4.43 mg/L) + Kin (4.64 mg/L) + IAA (5.7 mg/L)	5–7	+++
IAA (28.54 mg/L)	−	−
2,4-D (22.62 mg/L)	5–7	+++
6-BA (22.19 mg/L)	10–12	++
Kin (23.23 mg/L)	10–12	+

IAA, indole-3-acetic acid; 2,4-D = 2,4-dicholorophenoxyacetic acid; 6-BA = 6-benzyladenine; Kin = kinetin.

^a.+++, prominent; ++, moderate; +, feeble; −, no inititation.

Figure 2 Initiation of callus from leaf explant of germinated seedling after 15 days in MS medium supplemented with 2,4-D (4.52 mg/L) + 6-BA (4.43 mg/L).

Table 2. Effects of plant growth regulators on development of callus in MS medium.

S. no.	Growth regulators (µM)	Extent of callus development^a.	Nature of calli
1	2,4-D (13.58 mg/L)	++	Brown hard callus
2	2,4-D (13.58 mg/L) + Kin (4.64 mg/L)	++	Yellowish soft callus
3	2,4-D (4.52 mg/L) + 6-BA (4.43 mg/L)	+++	Yellowish soft and friable callus
4	2,4-D (4.52 mg/L) + 6-BA (4.43 mg/L) + Kin (4.64 mg/L) + IAA (5.7 mg/L)	+++	Yellowish soft and friable callus
5	2,4-D (22.62 mg/L)	+++	Brown hard callus
6	6-BA (4.43 mg/L) + Kin (4.64 mg/L)	++	Yellowish friable callus
7	2,4-D (4.52 mg/L) + IAA (5.7 mg/L)	+	Yellowish friable callus

IAA, indole-3-acetic acid; 2,4-D = 2,4-dicholorophenoxyacetic acid; 6-BA = 6-benzyladenine; Kin = kinetin.

^a.+++, prominent; ++, moderate; +, poor.

Figure 3 Developed callus from leaf explant of germinated seedling after 45 days in MS medium supplemented with 2,4-D (4.52 mg/L) + 6-BA (4.43 mg/L) + ascorbic acid (1 g/L).

Figure 4 Transverse section of liver showing normal architecture of hepatocytes.

Figure 5 Transverse section of liver showing necrosis with fatty changes due to PCM toxicity.

Figure 6 Leaf callus extract–treated liver section showing recovery from PCM toxicity.

Figure 7 Wedelolactone-treated liver section showing recovery from PCM toxicity.

Figure 8 Silymarin-treated liver section showing recovery from PCM toxicity.

Table 3. Standard plot data for estimation of paracetamol by spectrophotometric method.

Concentration of paracetamol (µg/ml)	Absorbance (at 510 nm)
0.0	0.0
1.0	0.013
2.0	0.027
4.0	0.053
6.0	0.079
8.0	0.106
10.0	0.132
12.0	0.159
14.0	0.185

Table 4. Antihepatotoxic effects of ethanol extract of plant leaves, leaf callus, wedelolactone, demethylwedelolactone, and silymarin on biochemical parameters in paracetamol-induced hepatic injury.

Group	SGPT (units/ml)	SGOT (units/ml)	Serum albumin	Serum total protein	Serum alkaline phosphatase	T. Bil. (mg/dl)
Group I (normal)	35.5 ± 2.29	57.6 ± 4.06	3.85 ± 0.42	4.72 ± 0.31	18.17 ± 1.48	1.26 ± 0.06
Group II (Paracetamol control)	188.4 ± 6.18	156.6 ± 4.2	3.95 ± 0.48	4.94 ± 0.45	33.8 ± 2.6	1.86 ± 0.20
Group III (E. alba. plant leaves)	124.8 ± 4.64*	110.8 ± 3.18*	3.88 ± 0.54*	4.84 ± 0.62*	26.6 ± 2.2*	1.64 ± 0.26*
Group IV (E. alba. plant leaf callus)	114.2 ± 4.28*	108.8 ± 3.8*	3.89 ± 0.42*	4.83 ± 0.38*	25.8 ± 3.4*	1.63 ± 0.26*
Group V (Wedelolactone)	78.8 ± 2.26*	72.6 ± 2.48*	3.86 ± 0.38*	4.75 ± 0.56*	22.2 ± 2.0*	1.42 ± 0.08*
Group VI (Demethylwedelolactone)	93.8 ± 4.29*	84.6 ± 5.44*	3.87 ± 0.52*	4.78 ± 0.34*	24.2 ± 1.8*	1.54 ± 0.12*
Group VII (Silymarin)	84.8 ± 2.46*	76.6 ± 2.14*	3.86 ± 0.26*	4.76 ± 0.44*	22.4 ± 1.8*	1.46 ± 0.12*
Group II (After self-recovery)	146.2 ± 4.84	132.4 ± 4.32	3.90 ± 0.66	4.88 ± 0.58	27.8 ± 3.64	1.78 ± 0.24

Note.: Students t.-test was performed. Each value represents the mean SE for six rats.

SGPT, serum glutamyl pyruvate transaminese; SGOT, serum glutamyl oxaloacetate transaminese; T. Bil, total bilirubin.

*Significant difference from carbon-tetrachloride group (group II), p < 0.05.

Table 5. Peak plasma concentration (C_max), time of peak concentration (T_max), and area under the curve (AUC) of paracetamol.

S. no.	Time (h)	Peak plasma concentration (µg/ml) (normal control)	Peak plasma concentration (µg/ml) (paracetamol + wedelolactone)	Peak plasma concentration (µg/ml) (paracetamol + E. alba. leaf callus extract)	Peak plasma concentration (µg/ml) (paracetamol + E. alba. leaves extract)
1	0.5	11.25 ± 0.42	8.25 ± 0.35	7.9 ± 0.45	7.70 ± 0.80
2	1.0	20.30 ± 1.6	13.0 ± 1.24	14.5 ± 1.25	13.20 ± 1.45
3	2.0	31.20 ± 2.4	21.20 ± 1.42	23.45 ± 1.6	22.0 ± 1.85
4	3.0	16.10 ± 1.8	29.50 ± 1.46	32.20 ± 2.25	31.4 ± 2.0
5	4.0	8.25 ± 0.65	12.64 ± 1.16	12.65 ± 1.36	12.75 ± 0.80
6	5.0	4.30 ± 0.12	6.64 ± 0.44	7.15 ± 0.82	6.85 ± 0.62
7	6.0	2.30 ± 0.12	2.82 ± 0.22	3.10 ± 0.32	3.0 ± 0.38
AUC	—	77.0	87.2	86.0	84.6
Cmax	—	31.2	31.0	30.0	29.5
Tmax	—	2.0 h	3.0 h	3.0 h	3.0 h

Note.: values are mean of six animals.