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Sci Rep
2018 Jun 22;81:9506. doi: 10.1038/s41598-018-27932-x.
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Function of Thelenota ananas saponin desulfated holothurin A in modulating cholesterol metabolism.
Han QA
,
Li K
,
Dong X
,
Luo Y
,
Zhu B
.
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This work was designed to separate and purify the saponin from Thelenota ananas with the highest anti-cholesterol ability using multiple chromatography and mass spectrometry analyses, and to systematically investigate the effect of the Thelenota ananas saponin on cholesterol metabolism in oxidized low-density lipoprotein (ox-LDL) induced macrophage foam cells. Desulfated holothurin A (desHA), which was finally identified as the targeted saponin with the highest activity in decreasing low-density lipoprotein-cholesterol (LDL-C), markedly inhibited the formation of foam cells derived from macrophages based on Oil Red O staining. In addition, desHA significantly blocked the synthesis of fatty acid synthetase while promoted intracellular cholesterol efflux. Furthermore, desHA inhibited the effects of ox-LDL on macrophage mRNA expression, which enhanced the level of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoAR) and suppressed the expression of SR-BI, ABCA1 and ABCG1, which further increased the levels of extracellular cholesterol and triglyceride. Blocking AKT and AMPK pathway and LXR synthesis revealed that desHA also regulated the contents of HMG-CoAR and eNOS via LXR/AKT/AMPK pathway. Thus, desHA played an essential role in cholesterol efflux and synthesis, which indicated desHA and Thelenota ananas are valuable resources to exploit new functional food and nutraceuticals.
Figure 1. The collision-induced fragmentation patterns of compound 3 separated by semi-preparative HPLC. Blue and Black arrows are two possible fragmentation patterns. Note: Agl is short for Aglycone, MeGlc is short for 3-O-methylglucose, Glc is short for Glucose, Qui is short for Quinovose, Xyl is short for Xylose.
Figure 3. The effect of desHA on macrophage-derived foam cells formation with a magnification of ×400. (A) control group, (B) ox-LDL group, (C) ox-LDL + 1 µg/mL desHA group, (D) ox-LDL + 3 µg/mL desHA group, (E) ox-LDL + 5 µg/mL desHA group.
Figure 4. The effect of desHA on the productions of FAS (A), TG (B) and TC (C) in RAW264.7 cells.
Figure 5. The effect of desHA on the expressions of cholesterol metabolism related factors HMG-CoAR (A), SR-BI (B), ABCA1 (C) and ABCG1 (D).
Figure 6. The cell signal pathways mediated by desHA in cholesterol metabolism. (A) The changes of AKT, P-AKT, AMPK and P-AMPK levels treated by AKT inhibitor with or without desHA. (B) The changes of P-AMPK and its downstream protein factors treated by AMPK inhibitor with or without desHA. (C) The changes of P-AKT expression treated by LXR siRNA with or without desHA.
Figure 7. The molecular mechanism of desHA on regulating cholesterol metabolism in macrophage foam cells.
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