Study on the Viability Inhibitory Effects and Mechanism of Dictamnine on Mice Spleen Lymphocyte in vitro / 中国药房

ABSTRACT

OBJECTIVE： To study the inhibitory effect of dictamnine on the viability of mice spleen lymphocyte in vitro and explore its potential mechanism. METHODS： The primary spleen lymphocytes of mice were isolated and cultured. The cells were treated with 0 （blank control）， 50， 100， 150 μmol/L dictamnine for 24 h. MTT assay was used to determine the cell viability； Lactic dehydrogenase （LDH） assay was used to determine release rate of LDH. Early apoptosis rate was detected by flow cytometry. Necrosis rate was detected by Hoechst 33342 and PI double staining； Western blot assay was used to detect the protein expressions of Caspase 3 and Cleaved-Caspase 3 in cells. Comet assay was used to detect DNA damage in cells （reflected in the proportion of DNA tail area）. RESULTS： Compared with blank control， 100， 150 μmol/L dictamnine could significantly inhibit the viability of lymphocytes （P＜0.01）. 150 μmol/L dictamnine could significantly increase the release of LDH （P＜0.05）， and release rate reached 79.37％. 50， 100， 150 μmol/L dictamnine could improve the early apoptotic rate of lymphocyte， but there was no statistical significance （P＞0.05）. 150 μmol/L dictamnine could significantly increase the necrosis rate （P＜0.05）， and necrosis rate reached 78.64％. 50， 100， 150 μmol/L dictamnine could increase the protein expression of Caspase 3， but there was no statistical significance （P＞0.05）， while 50， 100 μmol/L dictamnine could improve the protein expression of Cleaved-Caspase 3 significantly （P＜0.05）. DNA damage was induced in a dose-dependent manner by dictamnine， in which 100 and 150 μmol/L dictamnine could significantly increase DNA tail area （P＜0.01）. CONCLUSIONS： Dictamnine can inhibit spleen lymphocyte viability， and the mechanism may be related to inducing spleen lymphocyte necrosis and DNA damage.