Study on Preparation of Avenanthramides and Mechanism of Avenanthramides-Induced Apoptosis of Lung Cancer Cells(燕麦蒽酰胺的制备及其促肺癌细胞凋亡机制研究)
摘要
Avenanthramides (Avns), the unique active ingredients in oat, have multiple physiological functions. However, the natural content of Avns is very low. In this work, a germination procedure was used to promote the biotransformation of Avns in oat grains in order to elevate its natural content, optimize the extraction conditions and analyze its benefit to health. The influence of different experimental parameters on extraction of Avns was carried out by a single factor experiment, and the extraction process was optimized using the response surface methodology. Finally, the potential mechanism of Avns-induced apoptosis of lung cancer cells was explored. Avns were extracted after oat germination using an ultrasonic-assisted traditional organic solvent extraction. High performance liquid chromatography (HPLC) was employed for qualitative and quantitative analysis. With Avn C as standard positive control, the Western blot was used to explore the regulating effect on expression of nuclear factor kappa B (NF-κB) related pathway proteins and X-linked inhibitor of apoptosis (XIAP). Flow cytometry analysis was performed to detect the apoptosis of non-small cell lung cancer (NSCLC) H1299 and A549 after Avns treatment. The results showed that the optimal extraction conditions for Avns yield were as follows:the solid-liquid ratio 1∶22.2g/mL, the ultrasonic power 87.6W, the ethanol volume fraction 84.2%. In this condition, the yield of Avns was 849.251μg/g. The data of Western blot and flow cytometry analysis showed that both Avn C and Avns down regulated NF-κB/p65 phosphorylation and XIAP expression, and exerted promoting effect on apoptosis of H1299 and A549 cells in a dose-dependent manner. The results indicated that Avns might induce apoptosis in lung cancer cells by inhibiting the activity of NF-κB/XIAP signal pathway. This study aimed to provide theoretical basis of the application of Avns for lung cancer prevention.(燕麦蒽酰胺(avenanthramides,Avns)作为燕麦中特有的活性成分,具有广泛的生理功能,但其天然含量较低