Label-free study of intracellular glycogen level in Metformin and Resveratrol-treated insulin-resistant HepG2 by live-cell FTIR spectroscopy

Anchisa Poonprasartporn; K. L. Andrew Chan

doi:https://doi.org/10.1016/j.bios.2022.114416

Label-free study of intracellular glycogen level in Metformin and Resveratrol-treated insulin-resistant HepG2 by live-cell FTIR spectroscopy

Anchisa Poonprasartporn, K. L. Andrew Chan^*

^*Corresponding author for this work

Institute of Pharmaceutical Science

Research output: Contribution to journal › Article › peer-review

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Abstract

Conventional in vitro study often involves the destruction of the cells followed by purification and dilution steps before applying enzymatic assay or metabolomic analysis. It is a costly and laborious process, and it cannot monitor changes as a function of time. Recently, we have developed a new label-free live-cell FTIR approach that can directly measure biochemical compositional changes within living cells in situ and the spectral changes are shown to be highly specific to the drug applied. In this work, we have demonstrated for the first time the effect of two anti-diabetic drugs, metformin and Resveratrol, on insulin-resistant liver cells (HepG2). Using live-cell FTIR with principal component analysis, we have shown the differences in the biochemical profiles between normal and insulin-resistant cells (p < 0.05), the lack of response/difference from the insulin-resistant cell to insulin (p > 0.05) and the restoration of the biochemical profile and sensitivity to insulin from the insulin-resistant cells after the drug treatment (p < 0.05). Particularly, a rise in the glycogen level, marked by three distinctive peaks at 1150, 1080 and 1020 cm ⁻¹, within the living cells after the anti-diabetic drug treatments is observed. The live-cell FTIR results are confirmed by a parallel gold-standard biochemical assay, demonstrating the restoration of insulin sensitivity of the insulin-resistance cells. Live-cell FTIR can be a complementary tool for drug efficacy screening, especially for insulin sensitizers.

Original language	English
Article number	114416
Journal	Biosensors and Bioelectronics
Volume	212
Early online date	27 May 2022
DOIs	https://doi.org/10.1016/j.bios.2022.114416
Publication status	Published - 15 Sept 2022

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title = "Label-free study of intracellular glycogen level in Metformin and Resveratrol-treated insulin-resistant HepG2 by live-cell FTIR spectroscopy",

abstract = "Conventional in vitro study often involves the destruction of the cells followed by purification and dilution steps before applying enzymatic assay or metabolomic analysis. It is a costly and laborious process, and it cannot monitor changes as a function of time. Recently, we have developed a new label-free live-cell FTIR approach that can directly measure biochemical compositional changes within living cells in situ and the spectral changes are shown to be highly specific to the drug applied. In this work, we have demonstrated for the first time the effect of two anti-diabetic drugs, metformin and Resveratrol, on insulin-resistant liver cells (HepG2). Using live-cell FTIR with principal component analysis, we have shown the differences in the biochemical profiles between normal and insulin-resistant cells (p < 0.05), the lack of response/difference from the insulin-resistant cell to insulin (p > 0.05) and the restoration of the biochemical profile and sensitivity to insulin from the insulin-resistant cells after the drug treatment (p < 0.05). Particularly, a rise in the glycogen level, marked by three distinctive peaks at 1150, 1080 and 1020 cm −1, within the living cells after the anti-diabetic drug treatments is observed. The live-cell FTIR results are confirmed by a parallel gold-standard biochemical assay, demonstrating the restoration of insulin sensitivity of the insulin-resistance cells. Live-cell FTIR can be a complementary tool for drug efficacy screening, especially for insulin sensitizers. ",

author = "Anchisa Poonprasartporn and Chan, {K. L. Andrew}",

note = "Funding Information: The increase of intracellular glycogen is an indicator of an increased glucose uptake when an insulin sensitizer was added. The mean integrated absorbance of the 1020 cm−1 peak between 1060 and 1000 cm−1 of the treatment and control has been plotted to semi-quantitatively approximate and compare the change in the glycogen level in cells over 24 h of treatment. The results have clearly shown that both metformin and Resveratrol treated cells have more glycogen compared to the control as presented in Fig. 4A. This integrated glycogen absorbance supports the PCA results that glycogen is being generated after the insulin sensitizer treatment (Fig. 3A3). Publisher Copyright: {\textcopyright} 2022 The Authors",

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AU - Chan, K. L. Andrew

N1 - Funding Information: The increase of intracellular glycogen is an indicator of an increased glucose uptake when an insulin sensitizer was added. The mean integrated absorbance of the 1020 cm−1 peak between 1060 and 1000 cm−1 of the treatment and control has been plotted to semi-quantitatively approximate and compare the change in the glycogen level in cells over 24 h of treatment. The results have clearly shown that both metformin and Resveratrol treated cells have more glycogen compared to the control as presented in Fig. 4A. This integrated glycogen absorbance supports the PCA results that glycogen is being generated after the insulin sensitizer treatment (Fig. 3A3). Publisher Copyright: © 2022 The Authors

PY - 2022/9/15

Y1 - 2022/9/15

N2 - Conventional in vitro study often involves the destruction of the cells followed by purification and dilution steps before applying enzymatic assay or metabolomic analysis. It is a costly and laborious process, and it cannot monitor changes as a function of time. Recently, we have developed a new label-free live-cell FTIR approach that can directly measure biochemical compositional changes within living cells in situ and the spectral changes are shown to be highly specific to the drug applied. In this work, we have demonstrated for the first time the effect of two anti-diabetic drugs, metformin and Resveratrol, on insulin-resistant liver cells (HepG2). Using live-cell FTIR with principal component analysis, we have shown the differences in the biochemical profiles between normal and insulin-resistant cells (p < 0.05), the lack of response/difference from the insulin-resistant cell to insulin (p > 0.05) and the restoration of the biochemical profile and sensitivity to insulin from the insulin-resistant cells after the drug treatment (p < 0.05). Particularly, a rise in the glycogen level, marked by three distinctive peaks at 1150, 1080 and 1020 cm −1, within the living cells after the anti-diabetic drug treatments is observed. The live-cell FTIR results are confirmed by a parallel gold-standard biochemical assay, demonstrating the restoration of insulin sensitivity of the insulin-resistance cells. Live-cell FTIR can be a complementary tool for drug efficacy screening, especially for insulin sensitizers.

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Label-free study of intracellular glycogen level in Metformin and Resveratrol-treated insulin-resistant HepG2 by live-cell FTIR spectroscopy

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