Title:Assessment of the Anti-adipogenic Effect of Crateva religiosa Bark Extract for Molecular Regulation of Adipogenesis: In Silico and In vitro Approaches for Management of Hyperlipidemia Through the 3T3-L1 Cell Line
Volume: 26
Issue: 5
Author(s): Monika Singh*, Monika Sachdeva and Nitin Kumar
Affiliation:
- Department of Pharmacology, I.T.S. College of Pharmacy, Ghaziabad, U.P., Affiliated with Dr. A.P.J. Abdul Kalam Technical University, Lucknow, India
Keywords:
3T3-L1 cell lines, in vitro, in silico, antihyperlipidemic, adipocyte, GC-MS analysis.
Abstract:
Aims: This study aimed to determine the phytoconstituents of Crateva religiosa bark
(CRB) and evaluate the hypolipidemic effect of bioactive CRB extract by preventing adipocyte
differentiation and lipogenesis.
Background: After performing the preliminary phytochemicals screening, the antioxidant activity
of CRB extracts was determined through a DPPH (2, 2-diphenyl-1-picrylhydrazyl) assay.
Ethyl acetate extract (CREAE) and ethanol extract (CRETE) of CRB were selected for chromatographic
evaluation.
Methods: The antihyperlipidemic potential was analyzed by molecular docking through the
PKCMS software platform. Further, a 3T3-L1 cell line study via in vitro sulforhodamine B assay
and western blotting was performed to confirm the prevention of adipocyte differentiation and
lipogenesis
Results: The total phenolic contents in CREAE and CRETE were estimated as 29.47 and 81.19
μg/mg equivalent to gallic acid, respectively. The total flavonoid content was found to be 8.78
and 49.08 μg/mg, equivalent to quercetin in CREAE and CRETE, respectively. CRETE exhibited
greater scavenging activity with the IC50 value of 61.05 μg/ mL. GC-MS analysis confirmed
the presence of three bioactive molecules, stigmasterol, gamma sitosterol, and lupeol, in CRETE.
Molecular docking studies predicted that the bioactive molecules interact with HMG-CoA reductase,
PPARγ, and CCAAT/EBP, which are responsible for lipid metabolism. In vitro, Sulforhodamine
B assays revealed that CRETE dose-dependently reduced cell differentiation and viability.
Cellular staining using ‘Oil Red O’ revealed a decreased lipid content in the CRETE-treated
cell lines. CRETE significantly inhibited the induction of PPARγ and CCAAT/EBP expression,
as determined through protein expression via western blotting.
Conclusion: The influence of CRETE on lipid metabolism in 3T3-L1 cells is potentially suggesting
a new approach to managing hyperlipidemia.
