High content screening for promoting effect of Dioscorea opposita on macrophage phagocytosis / 中草药

ABSTRACT

Objective: To establish a high content screening (HCS) method by testing the phagocytic function of RAW 264.7, and to evaluate the effect of Dioscorea opposita on cell proliferation and phagocytosis in vitro. Methods: CCK-8 was used to measure the proliferation of RAW 264.7, and HCS was helpful to determine the ability of RAW 264.7 to engulf GFP-E.coli. Results: The best method was established by examining the time of administration, the amount of bacteria added, and the time of bacterial stimulation administration time of 12 h, 50 times of bacteria, and stimulation time of 1.5 h. Then, the repeatability of this method was tested, and the RSD value was 2.31%. Compared with the group without drugs, at the concentration of 0.156-1.25 mg/mL, the water extract of D. opposita could promote the proliferation, and improve the average phagocytosis rate of every single RAW264.7 cell. And the samples had the strongest effect on proliferation and phagocytosis when the concentration was 1.25 mg/mL. Conclusion: An HCS method is established and firstly used in D. opposita. HCS has the advantage of accurate, intuitive, and high throughput. D. opposita can not only promote cell proliferation, but also improve the phagocytic ability of each single RAW264.7 in vitro. A new method is provided for further study on immune activity and on the mechanism of enhancing the immune activity that some tonic herbs may have, such as D. opposita.