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首页> 外文期刊>Journal of Pharmacological and Toxicological Methods >An optimized in vitro assay for screening compounds that stimulate liver cell glucose utilization with low cytotoxicity.
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An optimized in vitro assay for screening compounds that stimulate liver cell glucose utilization with low cytotoxicity.

机译:一种优化的体外测定方法,用于筛选可刺激肝细胞葡萄糖利用并具有低细胞毒性的化合物。

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摘要

INTRODUCTION: Screening compounds that stimulate liver cell glucose utilization with low cytotoxicity plays an important role in anti-diabetic drug discovery. Existing in vitro assay for the quantitative measurement of cell glucose utilization rate and cytotoxicity of compounds needs to be modified in order to improve the simplicity and reproducibility. An optimized assay was described to addresses these problems. METHODS: Compounds were directly mixed with cell suspension. After 20 h incubation, glucose decrement detection in culture medium was performed, followed by the assessment of cell viability using Cell Counting Kit-8 (CCK-8). RESULTS: A series of experiments were conducted to define optimal conditions such as cell inoculation density, incubation time and CCK-8 concentration in cell culture medium. Glucose utilization unit (GUU) was defined for the evaluation of both efficacy and cytotoxicity of a compound. It was found that Rosiglitazone significantly stimulated cell glucose utilization with negligible cytotoxicity at the concentration of 10(-5) M. CONCLUSION: The simplicity and efficiency of the optimized method has been proven in this study. Our findings show that this assay has potential application in anti-diabetic drug discovery such as new peroxisome proliferator-activated receptor gamma (PPARgamma) agonists.
机译:简介:筛选刺激肝细胞葡萄糖利用且细胞毒性低的化合物在发现抗糖尿病药物中起着重要作用。为了改进简单性和可重复性,需要修改用于定量测量细胞葡萄糖利用率和化合物细胞毒性的现有体外测定法。描述了优化的测定法以解决这些问题。方法:将化合物直接与细胞悬浮液混合。孵育20小时后,在培养基中进行葡萄糖减量检测,然后使用Cell Counting Kit-8(CCK-8)评估细胞活力。结果:进行了一系列实验以确定最佳条件,例如细胞接种密度,孵育时间和细胞培养基中CCK-8浓度。定义了葡萄糖利用单位(GUU)以评估化合物的功效和细胞毒性。发现罗格列酮在10(-5)M的浓度下可显着刺激细胞葡萄糖利用,而细胞毒性可忽略不计。结论:本研究证明了优化方法的简单性和效率。我们的发现表明,该测定法在抗糖尿病药物的发现中具有潜在的应用,例如新的过氧化物酶体增殖物激活的受体γ(PPARgamma)激动剂。

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