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Published on: April 23, 2018
Establishment of a cell-based assay to screen insulin-like hypoglycemic drugs
1Institute of Materia Medica, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing, China.
Abstract:
This study sought to establish a cell based assay to screen insulin analogs. Previous studies have proposed that up-regulation of glucose consumption may have the same anti-diabetic effects as insulin. Here, the amount of glucose that disappeared in culture medium after incubation with insulin or drugs was determined and served as an indicator of the glucose consumption of the cells. In order to establish a cellular model to screen insulin analogs, the sensitivities of four cell lines - BALB/c 3T3, HepG2, NIH3T3, and Bel7402 - to insulin were evaluated by detecting glucose consumption after incubation with insulin (0-125 nM) for 24 h. BALB/c 3T3 was more sensitive to insulin than the other three cell lines. Insulin elevated glucose consumption of BALB/c 3T3 in a concentrationand time- manner. Glucose consumption of BALB/c 3T3 increased by 30% after incubation with insulin (30 nM) for 24 h. Insulin increased the proliferation of BALB/c 3T3 at 48 h. A model was established by detecting glucose consumption after treating BALB/c 3T3 with drugs for 24 h. Using the cell-based assay, we screened more than two thousand samples from Traditional Chinese Medicine (TCM). Five extracts exhibiting glucose absorbance in medium were identified, indicating a hit rate of 0.5%. Results suggested that a cell-based assay by detection of glucose consumption in BALB/c 3T3 was suitable for high-throughput screening and was feasible to identify insulin-like hypoglycemic drugs. Five hits were discovered from natural products. Further characterization of these active extracts could help to identify potential anti-diabetic drugs.
Insights
A new cell-based assay using BALB/c 3T3 cells effectively screens for insulin analogs by measuring glucose consumption. This high-throughput method identified five potential anti-diabetic compounds from Traditional Chinese Medicine extracts.
Area of Science:
- Biochemistry
- Pharmacology
- Cell Biology
Background:
- Up-regulation of glucose consumption is a proposed mechanism for anti-diabetic effects, similar to insulin.
- A cell-based assay is needed to screen for insulin analogs and identify novel hypoglycemic agents.
Purpose of the Study:
- To establish and validate a cell-based assay for screening insulin analogs and insulin-like hypoglycemic drugs.
- To evaluate the sensitivity of different cell lines to insulin and select the most responsive one for the assay.
Main Methods:
- Assessed insulin sensitivity of four cell lines (BALB/c 3T3, HepG2, NIH3T3, Bel7402) based on glucose consumption.
- Optimized the cell-based assay using BALB/c 3T3 cells by determining optimal insulin concentration and incubation time.
- Screened over two thousand Traditional Chinese Medicine (TCM) samples using the established cell-based assay.
Main Results:
- BALB/c 3T3 cells demonstrated higher sensitivity to insulin compared to other cell lines.
- Insulin significantly increased glucose consumption in BALB/c 3T3 cells in a dose- and time-dependent manner.
- The cell-based assay successfully identified five TCM extracts with significant glucose-lowering effects, yielding a hit rate of 0.5%.
Conclusions:
- A cell-based assay measuring glucose consumption in BALB/c 3T3 cells is suitable for high-throughput screening of insulin-like hypoglycemic drugs.
- The assay is feasible for identifying potential anti-diabetic agents from natural products.
- Further investigation of the five identified TCM extracts may lead to the discovery of new anti-diabetic drugs.
