Astaxanthin Counteracts Vascular Calcification In Vitro Through an Early Up-Regulation of SOD2 Based on a Transcriptomic Approach

Abstract

Vascular calcification (VC) is a critical contributor to the rising cardiovascular risk among at-risk populations such as those with diabetes or renal failure. The pathogenesis of VC involves an uprising of oxidative stress, for which antioxidants can be theoretically effective. However, astaxanthin, a potent antioxidant, has not been tested before for the purpose of managing VC. To answer this question, we tested the efficacy of astaxanthin against VC using the high phosphate (HP)-induced vascular smooth muscle cell (VSMC) calcification model. RNAs from treated groups underwent Affymetrix microarray screening, with intra-group consistency and inter-group differential expressions identified. Candidate hub genes were selected, followed by validation in experimental models and functional characterization. We showed that HP induced progressive calcification among treated VSMCs, while astaxanthin dose-responsively and time-dependently ameliorated calcification severities. Transcriptomic profiling revealed that 3491 genes exhibited significant early changes during VC progression, among which 26 potential hub genes were selected based on closeness ranking and biologic plausibility. SOD2 was validated in the VSMC model, shown to drive the deactivation of cellular senescence and enhance antioxidative defenses. Astaxanthin did not alter intracellular reactive oxygen species (ROS) levels without HP, but significantly lowered ROS production in HP-treated VSMCs. SOD2 knockdown prominently abolished the anti-calcification effect of astaxanthin on HP-treated VSMCs, lending support to our findings. In conclusion, we demonstrated for the first time that astaxanthin could be a potential candidate treatment for VC, through inducing the up-regulation of SOD2 early during calcification progression and potentially suppressing vascular senescence.

Role of Uremic Toxins in Early Vascular Ageing and Calcification

“…Astaxanthin, is a carotenoid derivative found in the red pigments of crustacean shells and salmons with known potent antioxidative effect. High phosphate concentration treated VSMCs showed reduced pro-calcifying properties when exposed to astaxanthin [ 199 ]. Mitoquinone (a mitochondria-targeting antioxidant formed by covalently binding ubiquinone or coenzyme Q) attenuated VC by suppressing oxidative stress and apoptosis of VSMCs through the Keap1/Nrf2 pathway [ 200 ].…”

Deep Learning-Assisted Repurposing of Plant Compounds for Treating Vascular Calcification: An In Silico Study with Experimental Validation

“…We harnessed an experimental model of VC established previously [ 10 , 11 , 15 ] for testing identified compounds. Briefly, a VSMC cell line, A7r5, between 3 and 6 passages was obtained from American Type Culture Collection, plated onto 24- and 6-well plates, and cultured with Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum (FBS), streptomycin/penicillin in 5% ambient CO 2 under 37°C.…”

“…Druggable targets inherent to the pathogenic landscape of VC can also be discovered through similar strategies. Using a transcriptomic approach followed by experimental validation, we previously showed that astaxanthin might beneficially influence VC severity through its action toward superoxide dismutase 2 (SOD2) [ 15 ]. Machine learning, a new strategy and expanding research discipline that trains computers to learn from massive data, emerges as a promising tool to help mine patterns or linkages [ 16 ].…”

Superoxide Dismutase 2 (SOD2) in Vascular Calcification: A Focus on Vascular Smooth Muscle Cells, Calcification Pathogenesis, and Therapeutic Strategies

“…Magnesium has also been shown to ameliorate oxidative stress and decrease vascular calcification severity through its effect on multiple pathways, including SM22 regulation [ 68 ]. Finally, using an in vitro model of VSMC calcification in combination with transcriptomic profiling, our research group recently discovered that astaxanthin, a carotenoid derivative with strong antioxidative property, exhibited a prominent anticalcific effect through specifically targeting SOD2 during treatment of calcified VSMCs [ 69 ]. Indeed, there are a handful of natural products or extracts exhibiting in vitro or in vivo effects of augmenting Sod2 expression and activities [ 70 , 71 ], and testing these SOD2-targeted candidates in VSMCs regarding their anticalcific potential may be a fruitful approach for identifying new pharmaceuticals for treating vascular calcification.…”