Acanthaster planci Inhibits PCSK9 and Lowers Cholesterol Levels in Rats

Abstract

Atherosclerosis is the main cause of cardiovascular diseases which in turn, lead to the highest number of mortalities globally. This pathophysiological condition is developed due to a constant elevated level of plasma cholesterols. Statin is currently the widely used treatment in reducing the level of cholesterols, however, it may cause adverse side effects. Therefore, there is an urgent need to search for new alternative treatment. PCSK9 is an enzyme responsible in directing LDL-receptor (LDL-R)/LDL-cholesterols (LDL-C) complex to lysosomal degradation, preventing the receptor from recycling back to the surface of liver cells. Therefore, PCSK9 offers a potential target to search for small molecule inhibitors which inhibit the function of this enzyme. In this study, a marine invertebrate Acanthaster planci, was used to investigate its potential in inhibiting PCSK9 and lowering the levels of cholesterols. Cytotoxicity activity of A. planci on human liver HepG2 cells was carried out using the MTS assay. It was found that methanolic extract and fractions did not exhibit cytotoxicity effect on HepG2 cell line with IC50 values of more than 30 µg/mL. A compound deoxythymidine also did not exert any cytotoxicity activity with IC50 value of more than 4 µg/mL. Transient transfection and luciferase assay were conducted to determine the effects of A. planci on the transcriptional activity of PCSK9 promoter. Methanolic extract and Fraction 2 (EF2) produced the lowest reduction in PCSK9 promoter activity to 70 and 20% of control at 12.5 and 6.25 μg/mL, respectively. In addition, deoxythymidine also decreased PCSK9 promoter activity to the lowest level of 60% control at 3.13 μM. An in vivo study using Sprague Dawley rats demonstrated that 50 and 100 mg/kg of A. planci methanolic extract reduced the total cholesterols and LDL-C levels to almost similar levels of untreated controls. The level of serum glutamate oxalate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) showed that the administration of the extract did not produce any toxicity effect and cause any damage to rat liver. The results strongly indicate that A. planci produced a significant inhibitory activity on PCSK9 gene expression in HepG2 cells which may be responsible for inducing the uptake of cholesterols by liver, thus, reducing the circulating levels of total cholesterols and LDL-C. Interestingly, A. planci also did show any adverse hepato-cytotoxicity and toxic effects on liver. Thus, this study strongly suggests that A. planci has a vast potential to be further developed as a new class of therapeutic agent in lowering the blood cholesterols and reducing the progression of atherosclerosis.

Acanthaster planci Inhibits PCSK9 Gene Expression via Peroxisome Proliferator Response Element (PPRE) and Activation of MEK and PKC Signaling Pathways in Human Liver Cells

“…During the preliminary screening, ten enhance fractions (Fraction 1–10) obtained through the process described in Section 4.1 were tested using gene reporter assay (Dual-Glo Luciferase Assay, Promega) to determine the effects of A. planci on the transcriptional activity of PCSK9 promoter. Fraction 2 produced the lowest reduction in PCSK9 promoter activity to 20% of control at 6.25 μg/mL among the ten fractions [ 35 ]. In addition, after preliminary screening, the further fractionation of Fraction 2 using column chromatography revealed the presence of a major compound that, upon nuclear magnetic resonance (NMR) analysis, was identified as deoxythymidine [ 35 ].…”

“…Fraction 2 produced the lowest reduction in PCSK9 promoter activity to 20% of control at 6.25 μg/mL among the ten fractions [ 35 ]. In addition, after preliminary screening, the further fractionation of Fraction 2 using column chromatography revealed the presence of a major compound that, upon nuclear magnetic resonance (NMR) analysis, was identified as deoxythymidine [ 35 ]. Therefore, Fraction 2 (selected fraction) was used for further studies presented in this study.…”