Abstract
Ginkgo biloba seeds are wildly used in the food and medicine industry. It has been found that 4′-O-methylpyridoxine (MPN) is responsible for the poisoning caused by G. biloba seeds. The objective of this study was to explore and optimize the extraction method of MPN from G. biloba seeds, and investigate its toxic effect on human gastric epithelial cells (GES-1) and the potential related mechanisms. The results showed that the extraction amount of MPN was 1.933 μg/mg, when extracted at 40 °C for 100 min, with the solid–liquid ratio at 1:10. MPN inhibited the proliferation of GES-1 cells, for which the inhibition rate was 38.27% when the concentration of MPN was 100 μM, and the IC50 value was 127.80 μM; meanwhile, the cell cycle was arrested in G2 phase. High concentration of MPN (100 μM) had significant effects on the nucleus of GES-1 cells, and the proportion of apoptotic cells reached 43.80%. Furthermore, the Western blotting analysis showed that MPN could reduce mitochondrial membrane potential by increasing the expression levels of apoptotic proteins Caspase 8 and Bax in GES-1 cells. In conclusion, MPN may induce apoptosis in GES-1 cells, which leads to toxicity in the human body.
Highlights
College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, Jiangsu, China; College of Food Engineering, Xuzhou University of Technology, Xuzhou 221018, Jiangsu, China; Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, Nanjing 210037, Jiangsu, China
The results indicated that a high concentration of MPN can have an effect on the fluorescence intensity of GES-1 cells’ nuclei after staining
When the concentration of MPN continued to increase, the expression level of Bax increased gradually. These results revealed that the expression levels of Caspase 8 and Bax in GES-1 cells can be increased by MPN, which can activate the apoptosis pathway in the cell
Summary
College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, Jiangsu, China; College of Food Engineering, Xuzhou University of Technology, Xuzhou 221018, Jiangsu, China; Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, Nanjing 210037, Jiangsu, China. Ginkgo biloba seeds are wildly used in the food and medicine industry. It has been found that 40 -O-methylpyridoxine (MPN) is responsible for the poisoning caused by G. biloba seeds. The objective of this study was to explore and optimize the extraction method of MPN from G. biloba seeds, and investigate its toxic effect on human gastric epithelial cells (GES-1) and the potential related mechanisms. High concentration of MPN (100 μM) had significant effects on the nucleus of GES-1 cells, and the proportion of apoptotic cells reached 43.80%. The Western blotting analysis showed that MPN could reduce mitochondrial membrane potential by increasing the expression levels of apoptotic proteins Caspase. MPN may induce apoptosis in GES-1 cells, which leads to toxicity in the human body. Key Contribution: The extraction method of MPN from Ginkgo biloba seeds was explored and optimized in this study.
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