Ethanol extract of Gynura bicolour reduces atherosclerosis risk by enhancing antioxidant capacity and reducing adhesion molecule levels

Figures & data

Table 1. Effects of GBEE on the growth characteristics and organ-related body weights of SD rats fed a HFD for 12 weeks.

	BW gain	Food intake	Heart	Liver	Kidney	Spleen
	g	g/day	% of BW
ND	331 ± 33^b	25.67	0.30 ± 0.03	3.17 ± 0.61	0.72 ± 0.05	0.17 ± 0.03
HFD	436 ± 71^a	20.43	0.28 ± 0.05	2.93 ± 0.23	0.65 ± 0.07	0.15 ± 0.02
LOVA	448 ± 56^a	21.38	0.26 ± 0.03	3.14 ± 0.38	0.61 ± 0.07	0.13 ± 0.01
GBEE-L	427 ± 66^a,b	21.81	0.27 ± 0.01	3.12 ± 0.28	0.66 ± 0.04	0.14 ± 0.02
GBEE-M	440 ± 20^a	21.93	0.28 ± 0.03	2.97 ± 0.39	0.65 ± 0.07	0.15 ± 0.04
GBEE-H	468 ± 83^a	21.55	0.27 ± 0.02	2.90 ± 0.17	0.63 ± 0.07	0.13 ± 0.01

SD rats fed a HFD were subdivided into the GBEE-L, GBEE-M, GBEE-H, and control groups and orally administered 4, 8 or 16 mg/kg BW GBEE or 40 mg/kg BW LOVA as a positive control, respectively. The data are expressed as the mean ± SD of three independent experiments.

^a,bValues of groups in the same cell phase marked with different letters differed significantly, as determined by Duncan’s test (p < 0.05).

Figure 1. Pathologic examination of the livers and thoracic aortas of SD rats treated with GBEE for 12 weeks. SD rats consuming a HFD were subdivided into GBEE-L, GBEE-M, and GBEE-H, and control groups and orally administered 4, 8, or 16 mg/kg BW GBEE or 40 mg/kg BW LOVA as a positive control, respectively. After H&E staining, pathologic examinations of the hepatic tissues (A) and the thoracic aorta (B) were performed. Fatty liver changes, steatosis, and endothelial inflammation are indicated by arrows.

Table 2. Effects of GBEE on TBARS and GSH levels in EA.hy926 cells.

	TBARS	GSH
	nmol/mg protein
Control	0.20 ± 0.01^b	51.6 ± 2.6^a
TNF-α (10 ng/mL)	0.48 ± 0.02^a	45.2 ± 3.6^b
TNF-α + GBEE (µg/mL)
10	0.36 ± 0.02^b	46.3 ± 2.7^a,b
50	0.29 ± 0.02^b	48.2 ± 2.4^a,b
100	0.27 ± 0.01^b	53.4 ± 5.1^a
GBEE (100 μg/mL)	0.19 ± 0.01^b	58.0 ± 4.2^a

EA.hy926 cells were pre-treated with 10, 50 or 100 μg/mL GBEE for 8 h and then stimulated with or without 10 ng/mL TNF-α for 3 h. EA.hy926 cells treated with alcohol served as a control group. The data are expressed as the mean ± SD of three independent experiments.

^a,bValues of groups in the same cell phase marked with different letters differed significantly, as determined by Duncan’s test (p < 0.05).

Table 3. Effects of GBEE on TBARS and GSH levels in the red blood cells of SD rats fed a HFD for 12 weeks.

	TBARS	GSH
	nmol/mg protein
ND	0.07 ± 0.01^b	0.36 ± 002^a
HFD	0.18 ± 0.02^a	0.25 ± 0.02^b
LOVA	0.11 ± 0.02^b	0.30 ± 0.05^a,b
GBEE-L	0.09 ± 0.02^b	0.32 ± 0.04^a,b
GBEE-M	0.07 ± 0.01^b	0.35 ± 0.01^a
GBEE-H	0.09 ± 0.01^b	0.38 ± 0.02^a

SD rats fed a HFD were subdivided into the GBEE-L, GBEE-M, GBEE-H, and control groups and orally administered 4, 8 or 16 mg/kg BW GBEE or 40 mg/kg BW LOVA as a positive control, respectively. The data are expressed as the mean ± SD of three independent experiments.

^a,bValues of groups in the same cell phase marked with different letters differed significantly, as determined by Duncan’s test (p < 0.05).

Figure 2. Effect of GBEE on the protein levels of E-selectin, ICAM-1, and VCAM-1 in EA.hy926 cells and SD rats. EA.hy926 cells were pre-treated with 10, 50 or 100 μg/mL GBEE for 8 h and then stimulated with or without 10 ng/mL TNF-α for 3 h. EA.hy926 cells treated with alcohol served as the control group. SD rats receiving a HFD were subdivided into the GBEE-L, GBEE-M, and GBEE-H, and control groups and orally administered 4, 8 or 16 mg/kg BW GBEE or 40 mg/kg BW LOVA as a positive control, respectively. Immunoblot assays were performed to analyze the expression levels of E-selectin, ICAM-1, and VCAM-1 in the EA.hy926 cells (A) and the thoracic aorta (C). Quantifications of the data are shown in (B) and (D). The data are expressed as the means ± SDs of three independent experiments. ^abThe values are significantly different those of the other groups, as determined by Duncan’s test (p < 0.05).

Figure 3. Analysis of the antioxidant activity of GBEE in vitro. (A) DPPH radical scavenging activity. (B) Reducing power. (C) Ferrous ion-chelating ability. (D) Superoxide radical-scavenging activity. The data are expressed as the means ± SDs of three individual experiments. ^abcdValues are significantly different from those of the other groups, as determined by Duncan’s test (p < 0.05). p-Values are <0.05 compared to the vitamin C, Na₂EDTA, or SOD standard groups.

Data availability statement

The datasets used and/or analysed during the present study are available from the corresponding author on reasonable request.