Cuban Sugar Cane Wax Alcohol Exhibited Enhanced Antioxidant, Anti-Glycation and Anti-Inflammatory Activity in Reconstituted High-Density Lipoprotein (rHDL) with Improved Structural and Functional Correlations: Comparison of Various Policosanols

Abstract

Policosanols from various sources, such as sugar cane, rice bran, and insects, have been marketed to prevent dyslipidemia, diabetes, and hypertension by increasing the blood high-density lipoproteins cholesterol (HDL-C) levels. On the other hand, there has been no study on how each policosanol influences the quality of HDL particles and their functionality. Reconstituted high-density lipoproteins (rHDLs) with apolipoprotein (apo) A-I and each policosanol were synthesized using the sodium cholate dialysis method to compare the policosanols in lipoprotein metabolism. Each rHDL was compared regarding the particle size and shape, antioxidant activity, and anti-inflammatory activity in vitro and in zebrafish embryos. This study compared four policosanols including one policosanol from Cuba (Raydel® policosanol) and three policosanols from China (Xi’an Natural sugar cane, Xi’an Realin sugar cane, and Shaanxi rice bran). The synthesis of rHDLs with various policosanols (PCO) from Cuba or China using a molar ratio of 95:5:1:1 with palmitoyloleoyl phosphatidylcholine (POPC): free cholesterol (FC): apoA-I:PCO (wt:wt) showed that rHDL containing Cuban policosanol (rHDL-1) showed the largest particle size and the most distinct particle shape. The rHDL-1 showed a 23% larger particle diameter and increased apoA-I molecular weight with a 1.9 nm blue shift of the maximum wavelength fluorescence than rHDL alone (rHDL-0). Other rHDLs containing Chinese policosanols (rHDL-2, rHDL-3, and rHDL-4) showed similar particle sizes with an rHDL-0 and 1.1–1.3 nm blue shift of wavelength maximum fluorescence (WMF). Among all rHDLs, the rHDL-1 showed the strongest antioxidant ability to inhibit cupric ion-mediated LDL oxidation. The rHDL-1-treated LDL showed the most distinct band intensity and particle morphology compared with the other rHDLs. The rHDL-1 also exerted the highest anti-glycation activity to inhibit the fructose-mediated glycation of human HDL2 with the protection of apoA-I from proteolytic degradation. At the same time, other rHDLs showed a loss of anti-glycation activity with severe degradation. A microinjection of each rHDL alone showed that rHDL-1 had the highest survivability of approximately 85 ± 3%, with the fastest developmental speed and morphology. In contrast, rHDL-3 showed the lowest survivability, around 71 ± 5%, with the slowest developmental speed. A microinjection of carboxymethyllysine (CML), a pro-inflammatory advanced glycated end product, into zebrafish embryos resulted in severe embryo death of approximately 30 ± 3% and developmental defects with the slowest developmental speed. On the other hand, the phosphate buffered saline (PBS)-injected embryo showed 83 ± 3% survivability. A co-injection of CML and each rHDL into adult zebrafish showed that rHDL-1 (Cuban policosanol) induced the highest survivability, around 85 ± 3%, while rHDL-0 showed 67 ± 7% survivability. In addition, rHDL-2, rHDL-3, and rHDL-4 showed 67 ± 5%, 62 ± 37, and 71 ± 6% survivability, respectively, with a slower developmental speed and morphology. In conclusion, Cuban policosanol showed the strongest ability to form rHDLs with the most distinct morphology and the largest size. The rHDL-containing Cuban policosanol (rHDL-1) showed the strongest antioxidant ability against LDL oxidation, anti-glycation activity to protect apoA-I from degradation, and the highest anti-inflammatory activity to protect embryo death under the presence of CML.

Beneficial Effect of Cuban Policosanol on Blood Pressure and Serum Lipoproteins Accompanied with Lowered Glycated Hemoglobin and Enhanced High-Density Lipoprotein Functionalities in a Randomized, Placebo-Controlled, and Double-Blinded Trial with Healthy Japanese

“…In the current study, the significantly reduced extent of glycation in hemoglobin and VLDL/LDL agreed with previous reports [ 16 , 43 ] showing that Cuban policosanol-encapsulated rHDL exhibited potent anti-glycation activity against fructose-mediated glycation of HDL and apoA-I. Recently, an rHDL containing Cuban policosanol exerted anti-glycation activity to prevent proteolytic degradation of HDL and apoA-I, while rHDL containing Chinese policosanol did not show anti-glycation activity [ 48 ].…”

“…An rHDL containing Cuban policosanol (Raydel ® ) showed a larger particle size and more particle numbers than other rHDLs containing Chinese policosanol. The rHDL-containing Cuban policosanol displayed potent anti-glycation activity to protect apoA-I and antioxidant activity to protect LDL, as reported recently [ 48 ]. The in vitro potentials of policosanol to enhance HDL functionality are linked with the in vivo efficacy in a human clinical study to improve blood pressure and dyslipidemia [ 12 ].…”

“…The CML also exhibited neurotoxicity and embryotoxicity in adult zebrafish and its embryo [ 47 , 48 ]. A microinjection of CML (final 500 ng) into zebrafish embryos caused acute embryo death, severe developmental defects, and the slowest developmental speed at 24 h post-injection [ 47 , 48 ].…”

“…The CML also exhibited neurotoxicity and embryotoxicity in adult zebrafish and its embryo [ 47 , 48 ]. A microinjection of CML (final 500 ng) into zebrafish embryos caused acute embryo death, severe developmental defects, and the slowest developmental speed at 24 h post-injection [ 47 , 48 ]. In the current study, a co-injection of HDL 2 from the policosanol group at week 12 showed the highest protection ability against embryonic death by neutralizing CML toxicity ( Figure 8 ).…”

“…Cuban policosanol consumption (10–20 mg/day) resulted in a remarkable decrease in LDL oxidation and HDL glycation in healthy subjects with pre-hypertension [ 18 ]. In vitro studies showed that rHDL containing Cuban policosanol exerted potent antioxidant, anti-glycation, and anti-inflammatory activities [ 19 , 43 , 48 ] with cholesterol efflux ability [ 19 ]. The binding ability of policosanol with apoA-I for discoidal rHDL formation is crucial for exerting physiological activities by maximizing the pluripotent functionality of HDL to prevent atherosclerosis, dyslipidemia, hypertension, and dementia [ 49 ].…”