EFEK AAPTAMINE TERHADAP EKSPRESI MRNA PPARG PADA DIFERENSIASI ADIPOSIT MENGGUNAKAN LINI SEL 3T3-L1
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Date
2023-07-14
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Abstract
Background and Objective: Regarding World Health Organization (WHO) data,
the number of obesity cases had tripled since 1975. The diversity of marine
environments, including aaptamine, has provided excellent prospects for human
living for a few years. Aaptamine was considered to have an adipogenic effect.
This study intends to investigate the inhibitory effect of aaptamine in inhibiting
mRNA expression of Pparg on adipocyte differentiation using 3T3-L1 cell line.
Materials and Methods: MTT Assay was performed to determine the non-toxic
concentration of aaptamine. Three sets of 3T3-L1 preadipocytes were prepared
with varying concentrations of aaptamine (0 µM, 25 µM, and 50 µM). On day 0,
each well received a cocktail for adipogenic differentiation containing
isobutylmethylxanthine (IBMX), dexamethasone, and insulin. Replacement of the
medium proceeded every two days. On day 13, the wells were harvested for
preparing qRT-PCR analysis. RNA was isolated using Quick-RNATM Miniprep
Kit and cDNA was synthesized from two microliters of total RNA using
SensiFASTTM cDNA Synthesis Kit. Afterward, qRT-PCR was performed using
the SensiFASTTM SYBR® No-ROX Kit based on the manufacturer’s instructions.
Results: The inhibition of Pparg mRNA expression by aaptamine at 25 M and 50
M (non-toxic range) concentrations in 3T3-L1 preadipocytes differed significantly
(p>0.05). Conclusion: Aaptamine inhibited the mRNA expression of Pparg
during the differentiation of 3T3-L1 cells.
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Keywords
Aaptamine, Lini Sel 3T3-L1, Ekspresi mRNA Pparg