Screening for autophagy modulators from an herbal extract library

• Main Authors: Shi-Zhen Liu, 劉士禎
• Other Authors: Chien-Yuan Pan
• Format: Others
• Language: en_US
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/53140686787057599236

碩士 === 國立臺灣大學 === 生命科學系 === 102 === Several herbal extracts (HEs), used in traditional Chinese medicine, have potential for the purpose of autophagy regulation. Autophagy is a lysosome-dependent degradation pathway. It plays important roles in starvation, stress response and apoptosis. To establish the platform to validate the modulators of autophagy, Irp62/HEK, which contain the constitutively expressed p62-luciferase for detection cells were used to detect the effects of herbal extracts on autophagy. Rapamycin was used as the activator control, and 3-methyladenine (3-MA) was used as the inhibitor control. Among the 497 HEs screened by the luciferase assay, 14 could serve as the activator of autophagy. Among 14 HEs, MTT assay was used to test the toxicity by measure the viability of the cells, and HE717 was chosen for the further study. The low toxicity of HE717 was carried out by trypan blue exclusion assay. The effect of activators was validated by Western blot of p62, flow cytometer measurement and fluorescence puncta of GFP-LC3. Taken together, HE717, isolated from whole plant of Lindernia sp., has the potential for drug development due to its activator effect of autophagy. This was the first platform to screen autophagy modulators from herbal library and the first report that Lindernia sp. could activate autophagy through p62 and LC3.