Institute of Pharmaceutical Science, School of Cancer and Pharmaceutical Sciences, King's College London, SE1 9NH, United Kingdom.
Institute of Pharmaceutical Science, School of Cancer and Pharmaceutical Sciences, King's College London, SE1 9NH, United Kingdom.
Biosens Bioelectron. 2022 Sep 15;212:114416. doi: 10.1016/j.bios.2022.114416. Epub 2022 May 27.
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, 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.
传统的体外研究通常涉及细胞的破坏，然后进行纯化和稀释步骤，再进行酶测定或代谢组学分析。这是一个昂贵且繁琐的过程，而且无法实时监测变化。最近，我们开发了一种新的无标记活细胞傅里叶变换红外（FTIR）方法，可以直接原位测量活细胞内的生化组成变化，并且所观察到的光谱变化高度特异于所应用的药物。在这项工作中，我们首次证明了两种抗糖尿病药物二甲双胍和白藜芦醇对胰岛素抵抗的肝细胞（HepG2）的作用。使用具有主成分分析的活细胞 FTIR，我们已经显示了正常和胰岛素抵抗细胞之间生化特征的差异（p<0.05），胰岛素抵抗细胞对胰岛素没有反应/差异（p>0.05），以及药物治疗后胰岛素抵抗细胞的生化特征和对胰岛素的敏感性恢复（p<0.05）。特别是，在用抗糖尿病药物治疗后，活细胞内的糖原水平升高，表现为 1150、1080 和 1020 cm 处三个特征峰。活细胞 FTIR 结果通过平行的黄金标准生化测定得到证实，证明了胰岛素抵抗细胞的胰岛素敏感性恢复。活细胞 FTIR 可以成为药物疗效筛选的补充工具，特别是对于胰岛素增敏剂。
