Biological effects of Byrsocarpus coccineus in vitro

Figures & data

short-legendFigure 1 

short-legendFigure 2 

Table 1.  Primers used for quantitative PCR.

IFNβ	5′-CAGCCGATTCATCGAGCACTCGC[FAM]G-3′
	5′-TTCCAGGACTGTCTTCAGATGG-3′
IFNα2A	5′-GACTCCCTCACAGCCAGAAATGGAG[FAM]C-3′
	5′-ATGACCTGGAAGCCTGTGTGat-3′

Table 2.  Effects of B. coccineus organic solvent extract fractions on cytochrome P450 activity in the BROD and BFCOD assays.

Treatment	Concentration (ng/mL)	BROD assay Specific activity (pmol/min/mg)	Concentration (ng/mL)	BFCOD assay Specific activity (pmol/min/mg)
Control	-	12.12 ± 0.51	-	160.4 ± 3.36
F6(2)	10^−4	12.64 ± 0.75	10^−3	257.3 ± 15.29^b
	10^−3	16.51 ± 1.05	10^−2	269.6 ± 7.78^b
	10^−2	23.6 ± 3.65^b	10^−1	287.7 ± 5.62^b
	10^−1	30.51 ± 5.06^b	1	283.0 ± 6.7^b
	1	31.84 ± 1.98^b	10	299.2 ± 5.72^b
F6(3)	10^−4	9.7 ± 0.9	10^−3	233.5 ± 2.4^b
	10^−3	11.71 ± 0.86	10^−2	262.6 ± 5.14^b
	10^−2	20.32 ± 3.87	10^−1	257.2 ± 5.37^b
	10^−1	24.07 ± 2.17^b	1	301.4 ± 3.96^b
	1	26.41 ± 4.28^b	10	303.2 ± 15.0^b
F(6)4	10^−4	10.48 ± 1.22	10^−3	317.9 ± 18.89^b
	10^−3	12.97 ± 1.32	10^−2	304.2 ± 3.45^b
	10^−2	21.02 ± 1.66	10^−1	265.9 ± 3.34^b
	10^−1	24.58 ± 1.6^b	1	257.6 ± 5.48^b
	1	26.91 ± 0.73^b	10	249.3 ± 4.4^b
Ketoconazole	(0.01 mM)	2.69 ± 0.31^a	(0.01 mM)	148.5 ± 1.29
	(0.1 mM)	-	(0.1 mM)	61.14 ± 6.34^b

Fractions F6(2), F6(3) and F6(4) were derived from organic solvent extraction of B. coccineus after the processes of maceration, liquid/ liquid extraction, silica gel and C-18 column chromatography as shown in Figure 1. Data are presented as mean ± SEM (n = 4).

^ap <0.05, ^bp <0.01 versus control (one way ANOVA followed by Dunnett’s post hoc test).

Table 3.  Effects of B. coccineus aqueous extract fractions on IFN expression.

PBMC	IFNβ	SEM	IFNα2a	SEM
control	1.0	0.2406	1.0	0.0787
BW 1	0.4961	0.0909	0.3807	0.0266
BW 2	1.3031	0.1923	0.2714	0.023
BW 3	0.4955	0.082	0.6425	0.0544
BW 4	0.6935	0.1396	0.6567	0.0593
BW 5	6.8148	0.9612	4.1267	0.3731
BW 6	4.5326	0.7374	1.6003	0.1515
BW 7	3.8798	0.4375	0.3639	0.0353
BW 8	4.8635	1.1508	0.2517	0.0194

Fractions BW1–BW8 were derived from the aqueous extract of B. coccineus after the processes of maceration and HP-20 column chromatography as shown in Figure 2. Data are presented as mean ± SEM. Bold type denotes fraction with the greatest effect selected for continued studies.

Table 4.  Effects of B. coccineus aqueous extract BW(5) fractions on IFNα expression.

PBMC	IFNα2a	SEM
control	1.0	0.0676
70% ETOH	0.3772	0.0404
BW CRUDE	2.8663	0.1572
BW(5)1	0.6225	0.0284
BW(5)2	4.9115	0.2681
BW(5)3	0.6206	0.0152
BW(5)4	26.4311	5.194
BW(5)5	2.1949	0.084
BW(5)6	1.2476	0.0862
BW(5)7	6.4382	0.4357
BW(5)8	0.8822	0.0508
BW(5)9	0.0715	0.0058
BW(5)10	0.0074	0.0013
BW(5)11	0.2907	0.0121
BW(5)12	0.2614	0.0113
BW(5)13	0.3507	0.0171
BW(5)14	0.4706	0.0229
BW(5)15	0.0201	0.0025
BW(5)16	3.6701	0.273
BW(5)17	0.0188	0.0025
BW(5)18	0.1127	0.0122
BW(5)19	0.0411	0.0072

Fractions BW(5)1-BW(5)19 were derived from the aqueous extract of B. coccineus after the processes of maceration, HP-20 column chromatography and HPLC at 300 nm using C-18 column gradient 20–100% MeOH as shown in Figure 2. Data are presented as mean ± SEM. Bold type denotes fraction with the greatest effect.

Table 5.  Effects of B. coccineus extract fractions on HT29 cell viability.

Concentra-tion (µg/mL)	% Cell viability
	F1		F2		F3		F4		F5		F6		F7		F8		F9		F10		F6(1)		F6(2)
1	112.2 ± 2.58		114.0 ± 1.0		113.3 ± 1.05		103.5 ± 3.13		102.4 ± 2.59		105.0 ± 4.09		106.8 ± 4.12		100.7 ± 4.59		95.82 ± 4.17		98.87 ± 1.28		107.5 ± 7.7		108.3 ± 6.04
10	67.38 ± 4.49		86.44 ± 3.05		70.36 ± 1.01		90.32 ± 5.97		93.39 ± 6.7		71.11 ± 12.38		85.55 ± 8.54		102.4 ± 2.41		102.4 ± 2.92		114.1 ± 14.98		101.5 ± 5.58		108.2 ± 7.76
100	34.7 ± 10.51		34.67 ± 5.53		36.69 ± 5.77		34.71 ± 14.03		20.56 ± 5.49		17.74 ± 5.27		22.24 ± 6.54		63.64 ± 8.63		109.5 ± 3.91		99.97 ± 6.52		103.6 ± 6.74		106.4 ± 4.7
Concentra-tion (µg/mL)	% Cell viability
	F6(3)	F6(4)		F6(5)		F6(6)		BW		BW1		BW2		BW3		BW4		BW5		BW6		BW7	BW8
1	105.0 ± 5.15	106.6 ± 6.84		102.7 ± 5.52		104.9 ± 3.38		108.9 ± 1.24		129.1 ± 1.11		136.7 ± 12.52		133.6 ± 2.06		131.8 ± 0.25		126.7 ± 0.55		110.6 ± 1.46		123.0 ± 5.92	120.1 ± 0.33
10	108.0 ± 5.09	104.3 ± 6.45		113.2 ± 5.46		102.7 ± 5.16		108.4 ± 2.54		128.9 ± 5.43		136.0 ± 3.7		138.8 ± 1.39		121.0 ± 0.16		119.7 ± 13.66		126.3 ± 0.53		126.5 ± 6.87	127.9 ± 2.04
100	103.0 ± 5.68	112.1 ± 2.28		56.94 ± 12.7		53.92 ± 5.37		102.1 ± 1.32		125.2 ± 1.86		121.5 ± 4.86		120.3 ± 0.83		106.2 ± 2.56		93.47 ± 2.11		98.14 ± 1.85		82.9 ± 5.32	125.4 ± 1.55

Fractions F1–F10 and F6(16) were derived from organic solvent extraction of B. coccineus after the processes of maceration, liquid-liquid extraction, silica gel and C-18 column chromatography as shown in Figure 1; Fractions BW(18) were derived from HP-20 column chromatography of the aqueous extract of B. coccineus (BW) as shown in Figure 2. Data are presented as mean ± SEM (n = 3).