Pharmacological evaluation for anti-asthmatic and anti-inflammatory potential of Woodfordia fruticosa flower extracts

Figures & data

Table 1. Percentage yield, phytochemical screening, and standardization of WFF extracts.

	Extracts obtained
Phytochemical tests	WFF-EA	WFF-AC	WFF-ME	WFF-HA
Weight of extracts (g) and % yield	17.24 (3.27)	12.58 (2.34)	20.26 (3.88)	8.46 (1.66)
Flavonoids	+	−	+	−
Alkaloids	−	−	−	−
Saponins	−	−	+	+
Tannins	−	+	+	+
Cardiac glycosides	+	−	−	−
Steroids	−	−	+	−
Triterpenoids	+	+	+	−
Carbohydrates	+	+	+	+
Standardization for content of phytochemicals
Total polyphenols	13.48 ± 1.02	47.66 ± 0.82	62.66 ± 1.89	56.00 ± 1.33
Total flavonoids	5.02 ± 0.29	ND	6.32 ± 0.76	ND
Total saponin	−	−	−	0.96 ± 0.17 ^a

For phytochemical screening: (+) presence of phytoconstituents and (–) absence of phytoconstituents; values of standardization for the content of phytochemical represent the mean ± SD of three independent replicates; ND, not determined due to absence of phytoconstituents in extracts.

^aDetermined for crude WFF powder.

Table 2. Effect of different WFF extracts on percentage DPPH inhibition, percentage metal chelation, percentage RRI of DPPH, reducing power, and percent histamine release.

			% RRI of DPPH				Reducing power (absorbances at 700 nm)					% Histamine release
	IC50 (µg/ml)		At time (h) for 100 µg/ml of treatment				Concentration of extracts (µg/ml)					Concentration of extracts (µg/ml)
Treatment	DPPH scavenging	Metal chelation	0.5	1	3	7	25	50	75	100	125	10	25	50	100
Standard	31.69	43.85	98.73 ± 1.44	83.45 ± 1.73	71.19 ± 0.78	59.16 ± 1.29	0.294 ± 0.076	0.413 ± 0.098	0.682 ± 0.128	0.841 ± 0.109	1.095 ± 0.153	97.56 ± 2.24	75.64 ± 2.86*	34.22 ± 2.10*	01.68 ± 1.07**
WFF-EA	44.44	38.12	78.24 ± 1.24	54.33 ± 0.86	38.66 ± 1.18	19.88 ± 1.45	0.291 ± 0.086	0.328 ± 0.092	0.419 ± 0.070	0.531 ± 1.10	0.765 ± 0.092	89.33 ± 1.09	46.67 ± 1.12*	13.57 ± 0.97*	07.12 ± 1.48**
WFF-ME	40.42	31.50	72.18 ± 1.18	57.43 ± 1.37	45.31 ± 1.73	25.37 ± 1.98	0.183 ± 0.067	0.349 ± 0.081	0.632 ± 1.08	0.747 ± 0.122	0.991 ± 0.093	77.39 ± 0.98*	33.42 ± 0.96*	12.78 ± 1.24*	06.52 ± 1.72**
WFF-AC	68.27	45.28	57.23 ± 2.03	25.41 ± 1.42	18.34 ± 1.12	06.73 ± 1.38	0.129 ± 0.034	0.273 ± 0.062	0.487 ± 0.09	0.591 ± 0.06	0.704 ± 0.10	96.85 ± 1.11	72.49 ± 0.93	45.39 ± 1.02	17.32 ± 1.93
WFF-HA	56.77	41.15	56.27 ± 1.65	42.71 ± 1.09	33.56 ± 1.39	21.92 ± 2.71	0.234 ± 0.17	0.395 ± 0.97	0.526 ± 0.12	0.796 ± 0.18	0.913 ± 1.02	80.33 ± 0.89*	58.67 ± 1.72*	19.86 ± 1.07*	10.13 ± 2.07*

All values represent the mean ± SD of three independent replicates. For percent histamine release determination, statistical significance was assessed using a one-way ANOVA with Bonferoni multiple comparison test. Ascorbic acid was used as a standard for percent DPPH inhibition, percentage RRI of DPPH, and reducing power; EDTA for percentage metal chelation and disodium cromoglycate (DSCG) for percent histamine release determination.

*p < 0.05, **p < 0.001.

Table 3. In vivo anti-inflammatory activity of different WFF extracts in egg albumin and carrageenan-induced inflammation model and in vitro heat and hypotonic solution induced hemolysis of erythrocyte membrane.

		% Inhibition of hemolysis		Measurement of paw volume and % inhibition of inflammation
				Egg albumin-induced inflammation (min)							Carrageenan-induced inflammation (h)
Treatment	Dose (mg/kg) & µg/ml	Heat-induced	Hypotonic solution-induced	0	20	40	60	80	100	120	0	01	03	05
Control	–	–	–	4.48 ± 0.10	5.45 ± 0.12	5.96 ± 0.23	6.62 ± 0.17	7.67 ± 0.21	7.30 ± 0.33	7.14 ± 0.19	4.20 ± 0.14	6.23 ± 0.13	7.45 ± 0.14	6.50 ± 0.19
WFF-EA	100	6.71 ± 1.52^ns	5.38 ± 1.25^ns	4.42 ± 0.21	5.40 ± 0.81 [0.91]	5.36 ± 0.77 [10.06]*	6.01 ± 0.61 [9.21]	5.98 ± 0.32 [22.03]*	5.67 ± 0.78 [22.32]*	6.37 ± 1.03 [10.78]*	4.43 ± 0.52	5.53 ± 0.87 [11.23]*	5.24 ± 0.65 [29.66]**	5.22 ± 0.32 [19.69]*
	200	18.92 ± 1.33*	16.31 ± 1.8*	4.30 ± 0.15	4.98 ± 0.72 [8.62]	5.72 ± 0.93 [4.02]	5.94 ± 0.82 [10.27]*	6.13 ± 0.50 [20.07]*	5.56 ± 0.97 [23.83]*	5.01 ± 0.63 [29.83]**	4.36 ± 0.36	5.86 ± 0.48 [5.93]	5.22 ± 0.81 [29.93]**	5.89 ± 0.45 [9.38]
WFF-ME	100	11.82 ± 1.77*	17.59 ± 1.07*	4.82 ± 0.13	5.33 ± 0.75 [2.20]	5.83 ± 0.56 [2.18]	5.14 ± 0.39 [22.35]*	6.27 ± 0.94 [18.25]*	5.86 ± 0.32 [19.72]*	6.43 ± 0.51 [10.08]*	4.68 ± 0.72	6.07 ± 0.24 [2.56]	5.88 ± 0.52 [21.07]*	5.16 ± 0.46 [20.61]*
	200	49.16 ± 2.48**	45.22 ± 1.46**	4.21 ± 0.17	4.84 ± 0.69 [11.19]*	5.51 ± 0.91 [7.55]	5.41 ± 0.43 [18.27]*	5.43 ± 0.38 [29.20]**	5.32 ± 0.61 [27.04]**	4.80 ± 0.75 [32.73]**	4.18 ± 0.26	5.06 ± 0.36 [18.78]*	4.58 ± 0.23 [38.52]**	4.39 ± 0.46 [32.46]**
WFF-HA	100	8.73 ± 1.32^ns	5.27 ± 2.15^ns	5.10 ± 0.12	5.30 ± 0.45 [2.75]	5.61 ± 0.66 [5.87]	5.57 ± 0.75 [15.86]*	6.78 ± 0.71 [11.60]	6.81 ± 1.12 [6.71]	6.92 ± 0.99 [3.08]	4.74 ± 0.64	6.17 ± 0.42 [0.96]	6.81 ± 0.86 [8.59]	5.66 ± 0.63 [12.92]*
	200	23.89 ± 1.96**	19.26 ± 1.83*	4.60 ± 0.18	5.13 ± 0.88 [5.87]	5.45 ± 0.67 [8.55]	5.62 ± 0.86 [15.10]*	5.76 ± 0.79 [26.07]**	5.94 ± 1.09 [18.63]*	5.23 ± 0.71 [26.75]**	4.38 ± 0.41	5.64 ± 0.23 [9.47]	5.85 ± 0.27 [21.47]*	5.20 ± 0.58 [20]*
Standard	10/50	19.46 ± 1.45*	18.13 ± 2.54*	4.12 ± 0.17	4.99 ± 0.13 [8.41]**	5.27 ± 0.12 [11.58]**	5.53 ± 0.23 [16.43]**	5.76 ± 0.16 [24.96]**	3.93 ± 0.18 [46.15]**	3.25 ± 0.14 [54.48]**	3.10 ± 0.12	5.05 ± 0.17 [18.97]*	3.15 ± 0.15 [57.72]**	3.67 ± 1.02 [43.53]**

For inhibition of hemolysis, values represent the mean ± SD of three replicates; standard used for percent inhibition of hemolysis was acetyl salicylic acid (50 µg/kg); treatment dose concentration used was µg/ml. For percent inhibition of inflammation, values represent the mean ± SD of six animals for each group; values in parenthesis indicate the percentage inhibition rate of inflammation, standard used was indomethacin (10 mg/kg) for egg albumin, and carrageenan-induced inflammation model, treatment dose concentration used was mg/kg; ns, non-significant.

*p < 0.01 and **p < 0.001 indicate different levels of statistically significant values against control.

Table 4. Evaluation of different WFF extracts and aminophylline for maximal bronchorelaxation in acetylcholine and histamine precontracted guinea pig tracheal chain and bronchoprotective effect on acetylcholine and histamine aerosol-induced bronchospasm in guinea pigs.

Bronchorelaxation in acetylcholine and histamine precontracted guinea pig tracheal chain													Bronchoprotective effect on acetylcholine and histamine aerosol-induced bronchospasm in guinea pigs
																Latency (s)
Treatment	Acetylcholine-induced contraction versus treatment in ml						Histamine-induced contraction versus treatment in ml						Treatment groups	Dose (mg/kg)	Tumble no.	Before treatment	After treatment	% Protection
groups	0.1	0.2	0.3	0.4	0.5	0.6	0.1	0.2	0.3	0.4	0.5	0.6	Control	–	14	88 ± 13	89 ± 14	–
Standard	18.17 ± 3.42	41.98 ± 2.75	68.25 ± 3.56	97.83 ± 3.09	97.83 ± 3.09	97.83 ± 3.09	18.82 ± 2.31	44.76 ± 3.78	78.25 ± 3.56	100 ± 3.85	100 ± 3.85	100 ± 3.85	Standard	10	08	90 ± 18	162 ± 27**	45.44
WFF-EA	11.32 ± 3.12	26.55 ± 4.06	44.87 ± 2.33	63.52 ± 2.65	80.95 ± 2.83	96.27 ± 2.88	18.7 ± 2.74	33.48 ± 3.57	58.93 ± 2.77	70.56 ± 3.02	90.23 ± 4.18	90.23 ± 4.18	WFF-EA	100	12	87 ± 14	113 ± 19	23.00
														200	08	81 ± 17	144 ± 16	43.75
WFF-ME	4.76 ± 1.34	19.04 ± 2.16	42.81 ± 3.02	66.66 ± 3.61	82.03 ± 3.34	100 ± 3.13	11.36 ± 3.12	36.73 ± 2.95	53.81 ± 2.83	81.68 ± 2.14	100 ± 1.94	100 ± 1.94	WFF-ME	100	09	79 ± 13	124 ± 09*	29.03
														200	05	85 ± 16	172 ± 22**	48.83
WFF-HA	9.38 ± 1.21	19.3 ± 1.48	44.89 ± 2.46	75.51 ± 4.13	91.81 ± 4.72	91.81 ± 3.96	7.35 ± 2.06	22.75 ± 1.31	42.05 ± 3.27	69.35 ± 2.43	84.56 ± 3.6	93.58 ± 4.12	WFF-HA	100	08	86 ± 21	119 ± 12	27.73
														200	06	92 ± 20	167 ± 21	44.91

For bronchorelaxation study, values represent the mean ± SD of four replicates for each group. *p < 0.01 versus acetylcholine- (at 0.4 ml of 1 µg/ml, 32 mm was considered 100%) or versus histamine (at 0.4 ml of 1 µg/ml, 29 mm was considered 100%)-induced contractions (a one-way ANOVA followed by post hoc Dunnette’s multiple comparison test. For bronchoprotective test, values represent the mean ± SD of six independent replicates.

*p < 0.05 and **p < 0.001 indicate the different levels of significance when compared against control group; standard – aminophylline.

Figure 1. Qualitative HPLC chromatogram for the determination of gallic acid in WFF-ME, depicting comparable retention and relative retention time.