Immunostimulant Activity of n-Butanol Fraction of Root Bark of Oroxylum indicum, vent.

Figures & data

FIG. 1 The structure of balcalein (aglucone of balcalin). Adapted from Lai et al. (2003).

TABLE 1 The percentage of baicalein in isolated fractions of O. indicum

Extraction solvent	Percent baicalein (w/v)
Petroleum ether	4.59%
Chloroform	0.48%
Ethyl Acetate	0.79%
n-Butanol	11.56%

TABLE 2 Effect of the n- butanol fraction (100 mg/kg/day per os) of root bark of O. indicum on antibody (Ab) formation against SRBC in sensitized rats (humoral immune response)

Group	Primary Ab titer on Day 13 (1° HA titer)	Secondary Ab titer on Day 20 (2° HA titer)
Control	3.58 ± 0.27	5.89 ± 0.29
n-Butanol	4.39 ± 0.22	7.51 ± 0.41
Dexamethasone	2.31 ± 0.29	3.12 ± 0.29

All values represent mean ± SEM, n = 6 in each group.

*p < 0.05, when compared with the control group (ANOVA, followed by Tukey's multiple range test), F_tab = 3.68; F_cal (2,15) = 15.22 (1°HA titer), 42.19 (2°HA titer). Dexamethasone treatment was at 0.27 mg/kg BW.

TABLE 3 Effect of n- butanol fraction (100 mg/kg/day per os) of root bark of O. indicum on SRBC-induced hind paw edema in pre-sensitized rats

Group	0 hr (on Day 20) Edema (mm)	48 hrs (on Day 22) Edema (mm)	% Edema
Control	2.50 ± 0.06	3.72 ± 0.11	49.04 ± 4.97
n-Butanol	2.50 ± 0.12	4.65 ± 0.25	87.01 ± 9.84
Dexamethasone	2.40 ± 0.06	3.16 ± 0.19	40.06 ± 4.20

All values represent mean ± SEM, n = 6 in each group.

*p < 0.05, when compared with the control group (ANOVA, followed by Tukey's multiple range test), F_tab = 3.68; F_cal(2,15) = 15.03 (% Edema). Dexamethasone treatment was at 0.27 mg/kg BW.

TABLE 4 Effect of the n- butanol fraction (100 mg/kg/day per os) of root bark of O. indicum on MDA content, anti-oxidant enzyme activities, and reduced glutathione levels

Treatment group	MDA content (μmole/mg protein)	SOD (mU/mg protein)	CAT (U/min/mg)	GSH (μmole/mg protein)
Control	0.682 ± 0.031	0.091 ± 0.004	0.655 ± 0.005	0.016 ± 0.000
n-Butanol	0.530 ± 0.029	0.220 ± 0.013	0.806± 0.028	0.095 ± 0.007
Dexamethasone	1.038 ± 0.040	0.064± 0.003	0.535 ± 0.016	0.025 ± 0.002

All values represent mean ± SEM, n = 6 in each group.

*p < 0.05, when compared with the control group (ANOVA, followed by Tukey's multiple range test), F_tab = 3.68; F_cal(2,15) = 59.35 (MDA), 111.30 (SOD), 51.79 (CAT), 96.81 (GSH). Dexamethasone treatment was at 0.27 mg/kg BW. Units for each enzyme are defined in the Methods section.

FIG. 2A Representative photomicrograph of splenic tissues recovered from rats on Day 22 of the respective indicated treatment regimens (WP = White pulp and RP = Red pulp)—Control rats. Left image at 10×, Right image at 40×.

FIG. 2B Representative photomicrograph of splenic tissues recovered from rats on Day 22 of the respective indicated treatment regimens (WP = White pulp and RP = Red pulp)—n-Butanol extract-treated rats. Left image at 10×, Right image at 40×.

FIG. 2C Representative photomicrograph of splenic tissues recovered from rats on Day 22 of the respective indicated treatment regimens (WP = White pulp and RP = Red pulp)—Dexamethasone-treated rats. Left image at 10×, Right image at 40×.

FIG. 3A Representative photomicrograph of thymus recovered from rats on Day 22 of the respective indicated treatment regimens (L = Lymphocytes, S = Sinusoid, HB = Hassel's Body. Left image at 10×, Right image at 40×.

FIG. 3B Representative photomicrograph of thymus recovered from rats on Day 22 of the respective indicated treatment regimens (L = Lymphocytes, S = Sinusoid, HB = Hassel's Body. (B) n-Butanol extract-treated rats; Left image at 10×, Right image at 40×.

FIG. 3C Representative photomicrograph of thymus recovered from rats on Day 22 of the respective indicated treatment regimens (L = Lymphocytes, S = Sinusoid, HB = Hassel's Body. (C) Dexamethasone-treated rats. Left image at 10×, Right image at 40×.

FIG. 4A Representative photomicrograph of axillary lymph node tissues recovered from rats on Day 22 of the respective indicated treatment regimens (C = Cortex, CP = Capsule, M = Medulla, and LF = Lymphoid follicles)—Control rats. Left image at 10×, Right image at 40×.

FIG. 4B Representative photomicrograph of axillary lymph node tissues recovered from rats on Day 22 of the respective indicated treatment regimens (C = Cortex, CP = Capsule, M = Medulla, and LF = Lymphoid follicles)—n-Butanol extract-treated rats. Left image at 10×, Right image at 40×.

FIG. 4C Representative photomicrograph of axillary lymph node tissues recovered from rats on Day 22 of the respective indicated treatment regimens (C = Cortex, CP = Capsule, M = Medulla, and LF = Lymphoid follicles)—Dexamethasone-treated rats. Left image at 10×, Right image at 40×.

TABLE 5 Effect of the n-butanol fraction (100 mg/kg/day per os) of root bark of O. indicum on alum adjuvant-induced hind paw edema in pre-sensitized rats

Treatment	0 hr (on Day 5) Edema (mm)	48 hrs (on Day 7) Edema (mm)	% Edema
Control	2.40 ± 0.07	3.37 ± 0.25	40.04 ± 8.82
n-Butanol	2.43 ± 0.08	4.37 ± 0.21	80.81 ± 9.97

All values represent mean ± SEM, n = 6 in each group.

*p < 0.05, when compared with the control group (Unpaired Student's t-test).

Lai M., Hsiu S., Tsai S., Hou Y., Chao P. Comparison of metabolic pharmacokinetics of baicalin and baicalein in rats. J. Pharmacy Pharmacol. 2003; 55: 205–209, [CSA]

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