Table 1.
Primer sequences used for qPCR.
Table 2.
RGR of cells after treatment of wedelolactone.
Fig 1.
Wedelolactone activated PPARα and phospho-AMPK.
(A): HepG2 cells were treated with or without wedelolactone at different concentrations for 24 h before collection of cell proteins and western blotting. (B) and (C): Protein levels were standardized against those of GAPDH. Histogram panels represent the ratios of PPARα/GAPDH (B) and phospho-AMPK/total AMPK (C). These values were obtained by quantification of the intensity of each band of the western blot. The values are expressed as the mean ± SD of triplicate assays. *p<0.05, **p<0.01 vs. control group.
Fig 2.
Up-regulation of PPARα expression was reversed by pre-treatment of MK886.
HepG2 cells were pretreated with MK886 (15 μM) for 0.5 h before the administration of fenofibrate (100 μM) or wedelolactone (25 μM) for 24 h. Quantification of PPARα was normalized by GAPDH. The values are expressed as the mean ± SD of triplicate assays. ***p<0.001 vs. control, ###p<0.001 vs. fenofibrate alone, ^^^p<0.001 vs. wedelolactone alone.
Fig 3.
Wedelolactone altered the expression of genes related to lipid metabolism.
(A), (B), (C) and (D): Effect of wedelolactone on mRNA expression in HepG2 cells treated with or without wedelolactone for 24 h. Vehicle and experimental groups were supplemented concurrently with DMSO, fenofibrate (100 μM) or wedelolactone (10 μM, 25 μM, 50 μM). (E) and (F): HepG2 cells were pretreated with MK886 (15 μM) for 0.5 h before administration of fenofibrate (100 μM) and wedelolactone (25 μM) for 24 h. The values are expressed as the mean ± SD of triplicate assays. *p<0.05, **p<0.01, ***p<0.001 vs. control, ##p<0.01, ###p<0.001 vs. fenofibrate alone, ^p<0.05, ^^p<0.01 vs. wedelolactone alone.
Fig 4.
Some gene expression was not affected by wedelolactone treatment.
Effect of wedelolactone on gene expressions in HepG2 cells with or without treatment of wedelolactone for 24 h. Vehicle and test groups were concurrently supplemented with DMSO, fenofibrate (100 μM) or wedelolactone (10 μM, 25 μM, 50 μM). The values are expressed as the mean ± SD of triplicate assays.
Fig 5.
Effect of wedelolactone on plasma levels of TC and TG in mice.
The normal group underwent intraperitoneal administration of physiologic (0.9%) saline and the other groups were injected with Triton WR-1339. The normal group and Triton group were treated with distilled water by intragastric administration. Test groups were treated with fenofibrate (100 mg/kg) or wedelolactone (100 mg/kg) by intragastric administration, respectively. The values are shown as the mean ± SD from eight animals in each group. ###p<0.001, Triton group vs. normal group, ***p<0.001, test group vs. Triton group.
Fig 6.
Effects of wedelolactone on plasma levels of TC (A), TG (B), LDL-C (C), and ALT (D) in Syrian hamsters.
The normal group was fed a normal diet whereas the other groups were fed a HFD. HFD + xue and HFD + WDL groups were supplemented concurrently with xue (xuezhikang) (250 mg/kg) or wedelolactone (10 mg/kg, 25 mg/kg, 40 mg/kg) for 4 weeks. The values are shown as the mean ± SD from ten animals in each group. ##p<0.01, ###p<0.001 vs. normal group, *p<0.05, **p<0.01, ***p<0.001 vs. HFD group.
Fig 7.
Hepatic levels of TC (A) and TG (B) in Syrian hamsters.
The normal group was fed a normal diet whereas the other groups were fed a HFD. The HFD + xue and HFD + WDL groups were supplemented concurrently with xuezhikang (250 mg/kg) or wedelolactone (10 mg/kg, 25 mg/kg, 40 mg/kg). After 4 weeks, livers were homogenized and levels of TC and TG were measured. The values are shown as the mean ± SD from ten animals in each group. ###p<0.001 vs. normal group, *p<0.05, **p<0.01, ***p<0.001 vs. HFD group.
Fig 8.
Wedelolactone activated AMPK and PPARα in Syrian hamsters.
Treatment with xuezhikang (250 mg/kg) or wedelolactone (10 mg/kg, 25 mg/kg, 40 mg/kg) for four weeks significantly increased the level of phospho-AMPK (A, C) and PPARα (D, E) in liver tissues. Protein levels were standardized against GAPDH levels. The histograms represent the ratios of AMPK/GAPDH (B), phospho-AMPK/total AMPK (C) and PPARα/GAPDH (E). These values were obtained by quantification of the intensity of each band of the western blot. The values are expressed as the mean ± SD from three animals in each group. ##p<0.01 vs. normal group, *p<0.05, **p<0.01 vs. HFD group.
Fig 9.
Wedelolactone alleviated liver damage and lipid accumulation.
(A) Histopathologic sections of liver from Syrian hamsters with different treatments were stained with H&E. (B) Histopathologic sections of liver from hamsters in each group were stained with Oil Red O. The normal group was fed a normal diet whereas the other groups were fed a HFD. HFD + xue and HFD + WDL groups were supplemented concurrently with xuezhikang (250 mg/kg) or wedelolactone (10 mg/kg, 25 mg/kg, 40 mg/kg).
Fig 10.
Wedelolactone increased the activities of SOD and up-regulated the expression of PPARα in OA-induced lipid peroxidized HepG2 cells.
HepG2 cells were stimulated with our without OA (25 μM) and treated with or without wedelolactone at different concentrations for 24 h before collection of cell lysates for SOD detection (A) and cell proteins for western blotting determination (B). OA, F100, WDL10, WDL25 and WDL50 groups were stimulated simultaneously with OA and treated with dimethyl sulfoxide (DMSO), fenofibrate (100 μM), or wedelolactone at 10, 25 or 50 μM, respectively. The values are expressed as the mean ± SD of triplicate assays. #p<0.05, ###p<0.001 vs. control group, *p<0.05, **p<0.01, ***p<0.001 vs. OA group.
Table 3.
The antioxidant activity of wedelolactone in vitro.
Fig 11.
Hepatic levels of MDA (A), SOD (B) and GSH-Px (C).
The normal group was fed a normal diet whereas the other groups were fed a HFD. HFD + xue and HFD + WDL groups were supplemented concurrently with xuezhikang (250 mg/kg) or wedelolactone (10 mg/kg, 25 mg/kg, 40 mg/kg). The values are shown as the mean ± SD from ten animals in each group. ###p<0.001 vs. normal group, **p<0.01, ***p<0.001 vs. HFD group.