Table 1.
Fatty acid composition of cocoa butter and refined and virgin pumpkin seed oil.
Table 2.
Phytochemical content of cocoa butter and refined and virgin pumpkin seed oil
Fig 1.
Refined and virgin pumpkin seed oils have beneficial effects on plasma lipids in cholesterol-fed ApoE*3Leiden mice.
Mice were fed a Western type diet (CON) containing cocoa butter (15% w/w of diet) for 20 weeks. The cocoa butter was in part replaced by refined pumpkin seed oil (REF) or virgin pumpkin seed oil (VIR) (each 9% w/w of diet). A: Plasma cholesterol levels over the course of the study, showing lower levels in REF and VIR-fed animals. B: Area under the curve analysis (AUC, expressed in arbitrary units; AU) of plasma cholesterol levels (t = 0 until t = 20 weeks) shows additional cholesterol-lowering effect of VIR compared with REF. C: Lipoprotein profile for cholesterol distribution in VLDL, LDL and HDL-sized particles shows cholesterol-lowering effect mainly confined to VLDL-sized particles. D: Plasma triglycerides over the course of the study were lowered by both REF and VIR. E: Area under the curve analysis of plasma triglyceride levels (t = 0 until t = 20 weeks) shows a reduction by VIR and REF. Data are mean±SEM. * p≤0.05, ** p≤0.01, *** p≤0.001 compared with CON. # p≤0.05, ## p≤0.01 for VIR compared with REF.
Fig 2.
Virgin pumpkin seed oil reduces circulating markers of inflammation in cholesterol-fed ApoE*3Leiden mice.
Mice were fed a Western type control diet (CON) or CON diet containing 9% refined pumpkin seed oil (REF) or 9% virgin pumpkin seed oil (VIR) for 20 weeks. A: Plasma SAA levels were reduced by VIR. Liver damage marker B: ASAT and C: ALAT were reduced by VIR but not by REF. D: Plasma sVCAM-1 levels in VIR animals were lower throughout the duration of the study. Data are mean±SEM. * p≤0.05, ** p≤0.01 compared with CON. # p≤0.05 for VIR compared with REF.
Fig 3.
Virgin pumpkin seed oil attenuates development of NAFLD in cholesterol-fed ApoE*3Leiden mice.
Mice were fed a Western type control diet (CON) or CON diet containing 9% refined pumpkin seed oil (REF) or 9% virgin pumpkin seed oil (VIR) for 20 weeks. A: Liver weight (expressed as percentage of terminal body weight) was reduced by REF and VIR. B: representative photomicrographs of HE-stained liver sections show presence of micro- (grey arrows) and macro- (white arrows)vesicular steatosis and inflammatory cell clusters (black arrows) in CON-fed animals, which was less pronounced in REF and more strongly reduced in VIR. C: Histological quantitative scoring of macrovesicular steatosis showed significant reduction in VIR. D: Hepatic triglyceride levels (biochemically determined) were reduced in both REF and VIR while only VIR significantly reduced E: hepatic cholesteryl ester content and F: free (unesterified) cholesterol levels. G: Histological quantification of number of inflammatory cell aggregates revealed a significant attenuation of hepatic inflammation by VIR. Data are mean±SEM. * p≤0.05, ** p≤0.01, *** p≤0.001 compared with CON. # p≤0.05, ### p≤0.001 for VIR compared with REF.
Fig 4.
Atherosclerosis development is reduced with virgin pumpkin seed oil.
Mice were fed a Western type control diet (CON) or CON diet containing 9% refined pumpkin seed oil (REF) or 9% virgin pumpkin seed oil (VIR) for 20 weeks. A: representative photomicrographs of HPS-stained cross sections of the aortic root show pronounced development of atherosclerosis in CON animals, which was less pronounced in REF and strongly reduced by VIR. B: Morphometric analysis of lesion area revealed a significant decrease in atherosclerotic lesion area by REF and VIR. C: Anti-atherogenic effects of pumpkin seed oils are specific on severe lesion types. D: Average lesion size was reduced in REF and VIR. E: Immunohistochemical staining for MAC-3 (CD107b) followed by quantification of positively stained area showed that both REF and VIR reduced the macrophage content of type V lesions. Data are mean±SEM. * p≤0.05, ** p≤0.01 compared with CON.
Fig 5.
Refined and virgin pumpkin seed oils modulate lipid metabolism and inflammatory gene expression.
Mice were fed a Western type control diet (CON) or CON diet containing 9% refined pumpkin seed oil (REF) or 9% virgin pumpkin seed oil (VIR) for 20 weeks. A: Hepatic lipogenic gene expression (Srebf1, Fasn, Dgat1) was reduced in both REF and VIR. B: Hepatic expression of genes involved in fatty acid catabolism (Ppara, Cpt1a, Acox1) was upregulated in VIR and to a lesser extent in REF. C. Only VIR reduced hepatic expression of inflammatory genes (Ccl2, Tnf, IL1b). All gene expression data are expressed as fold-change relative to CON. Data are mean±SEM. * p≤0.05, ** p≤0.01, *** p≤0.001 compared with CON, # p≤0.05, ### p≤0.001 for VIR compared with REF.