Department of Biological Sciences
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Item Anti-obesity and anti-steatotic effects of bixin (apocarotenoid from bixa orellana l. Seeds): dose-dependent correlations with bioactivity(MDPI, 2024-12) Sharma, Pankaj Kumar; Deepa, P.R.Annatto, a tropical shrub from Central and South America and parts of India, contains Bixin, an apocarotenoid pigment. Bixin is conventionally used as a natural food colorant and is now receiving attention for its health-promoting nutraceutical properties, particularly in chronic diseases (metabolic syndrome—MetS—and cancers). This study investigates the dose-dependent anti-obesity and anti-steatotic effects of Bixin in in vitro cell culture models. The anti-adipogenic and anti-steatotic effects of Bixin were examined in well-established in vitro models of obesity and non-alcoholic fatty liver disease (NAFLD/steatosis) using 3T3-L1 preadipocytes (by a differentiation protocol) and HepG2cells (steatosis-induced with oleic acid), respectively. Bixin was administered in the following concentration range: 1 μg mL−1–20 μg mL−1 (obesity model) and 2.5 μg mL−1–10 μg mL−1 (NAFLD/steatosis model). The neutral lipid content was estimated by Oil Red O staining; ROS/RNS were quantified by 2′,7′-dichlorofluorescein diacetate (DCFDA) and nitrite assays; and malondialdehyde (MDA), the biochemical marker of lipid peroxidation, was assessed by TBARS assay. At lower concentrations, 5 μg mL−1 in steatotic cells and 10 μg mL−1 in matured adipocytes, Bixin significantly reduced lipid accumulation in both hepatocytes and adipocytes (p < 0.05), demonstrating its potential as an anti-steatotic and anti-obesity agent. This beneficial effect was correlated with a reduction in oxidative stress levels (decreased MDA and ROS/RNS levels). Strikingly, at higher concentrations (>10 μg mL−1), Bixin showed increased lipid accumulation and oxidative stress. Bixin exhibits anti-obesity and anti-steatotic effects at lower doses, which correlates with its antioxidant properties. However, its bioactivity is dose-dependent, meaning that at higher concentrations, it ceases to inhibit adipogenesis. This opposing response is accompanied by elevated oxidative stress levels, indicating a pro-oxidant effect at higher doses, which suggests its anti-cancer potential. The present study highlights the significance of dosage optimization of nutraceuticals and dietary ingredients with respect to their intended biological applications, such as MetS and cancer treatment.Item Anti-adipogenic and anti-steatotic potential of edible pigment bixin and annatto seed extracts: LC-MS based bioactive profiling and in vitro biochemical validation(Elsevier, 2025-02) Sharma, Pankaj Kumar; Deepa, P.R.The rising prevalence of metabolic syndrome (MetS), such as obesity and its hepatic complication, and non-alcoholic fatty liver disease (NAFLD), necessitates safe, effective, and protective interventions. Natural products, such as carotenoids, including bixin derived from annatto seeds, have emerged as promising candidates due to their multifaceted pharmacological properties. This study aimed to characterize the edible food pigment bixin and other co-existing bioactives in acetone- and ethyl lactate-extracts of Bixa Orellana L. seeds, followed by their anti-adipogenic and anti-steatotic assessments using in-vitro models of obesity and NAFLD. LC-MS analysis revealed the presence of various phytochemicals in the bixin rich solvent extracts. In vitro studies demonstrated differential and significant anti-adipogenic and anti-steatotic effects of bixin (pure pigment) and solvent extracts of annatto seeds (P < 0.05). The LC-MS profiling of annatto seed extracts revealed the presence of bixin and several bixinoids. Other key phytochemicals that were identified were eicosatrienoic acid, geranylgeraniol, hypolatein, δ-tocotrienol, caffeoyl acid derivative, and zeaxanthin, which were differentially abundant in each solvent extract. Bixin and coexisting bioactives in the annatto seed extracts demonstrated significant anti-oxidant, anti-inflammatory, and anti-lipidemic effects (P < 0.05) in the present in vitro MetS models. Further studies may be directed toward evaluating the nutraceutical potential of bixin in combination with the implicated phytochemicals in the extracts for treating metabolic disorders.