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3,5-Dimethoxy-4-benzoic acid (syringic acid) a natural phenolic acid reduces reactive oxygen species in differentiated 3T3-L1 adipocytes

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Abstract

Preadipocytes under nutrient excess mature to lipid-laden adipocytes that are hotspots for generation of reactive oxygen species (ROS) imbalance and oxidative stress. Syringic acid (SA), a natural phenolic acid, was evaluated for its in vitro antioxidant and ROS modulation during in matured 3T3-L1 adipocytes. Following 10 d, the SA-treated adipocytes were evaluated for the levels of glutathione (GSH) and antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT). The levels of peroxides in mature adipocytes were estimated using dichlorofluorescein (DCF) cleavage fluorescence. The level of NADPH oxidase 4 (NOX4) expression was also investigated following 10-d differentiation period. SA significantly improved the levels of GSH, SOD, and CAT in matured adipocytes. Reduction in ROS production levels was also witnessed by decrease in DCF cleavage. SA showed concentration-dependent inhibition of NOX4 by day 7 of adipogenesis when compared with differentiated and undifferentiated cells. Moreover, SA exhibited effective antioxidant and anti-radical scavenging activity. These results suggest that SA in addition to inhibiting adipogenesis can strongly reduce ROS stress in mature adipocytes by upregulating levels of intracellular antioxidants and decreasing levels of NOX4 in 3T3-L1 adipocytes.

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Abbreviations

ABTS:

2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)

AT:

adipose tissue

BDE:

bond dissociation enthalpy

CAT:

catalase

Cu/Zn-SOD:

copper-zinc SOD

DC:

differentiated control

DCHF-DA:

2′,7′-dichlorofluorescin diacetate

DCF:

2′,7′-dichlorofluorescein

DPPH:

2,2-diphenyl-1-picrylhydrazyl

EC-SOD:

extracellular SOD

GSH:

glutathione

GLUT4:

glucose transporter type 4

IL:

interleukin

OS:

oxidative stress

ROS:

reactive oxygen species

SA:

syringic acid

SOD:

superoxide dismutase

Mn-SOD:

manganese SOD

MCE:

mitotic clonal expansion

NO:

nitric oxide

NOX4:

nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase

NADH:

nicotinamide adenine dinucleotide

TG:

triglyceride

UC:

undifferentiated control

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Acknowledgements

This research was conducted in the Department of Biomedical Sciences, Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, 600116.

Funding

This research was partially supported by Shri NPV Ramasamy Udayar PhD Research Scholarship.

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Authors and Affiliations

  1. Department of Biomedical Sciences, Sri Ramachandra Institute of Higher Education and Research, Chennai, Tamil Nadu, India

    Cordelia Mano John & Sumathy Arockiasamy

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  1. Cordelia Mano John
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  2. Sumathy Arockiasamy
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Correspondence to Sumathy Arockiasamy.

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Editor: Tetsuji Okamoto

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John, C.M., Arockiasamy, S. 3,5-Dimethoxy-4-benzoic acid (syringic acid) a natural phenolic acid reduces reactive oxygen species in differentiated 3T3-L1 adipocytes. In Vitro Cell.Dev.Biol.-Animal 57, 386–394 (2021). https://doi.org/10.1007/s11626-021-00549-7

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