Separation and purification and in vitro anti-proliferative activity of leukemia cell K562 of Galium aparine L. petroleum ether phase

Abstract

To explore material basis of in vitro anti-proliferative activity of leukemia cell K562 of petroleum ether phase of product resulting from Galium aparine L. 60% ethanol extraction, the experiment adopts column chromatography combined with thin layer preparation, isolates and purifies petroleum ether, conducts structural identification of obtained single compound and applies MTT method for viability assay of in vitro anti-proliferative activity of leukemia cell K562. Experimental results show that G. aparine L. petroleum ether contains mainly β-sitosterol, daucosterol and dibutyl phthalate and other substances. Under experimental conditions, the three could inhibit the proliferation of leukemia cell K562 with dose-effect and time-effect relationship, of which dibutyl phthalate has strongest activity. Dibutyl phthalate with excellent activity, β-sitosterol with rich content and moderate effect should be the main contributor to its biological activity.

Keywords: Galium aparine L.; K562; MTT; Purification.

Figure 1

Cytotoxicity of dibutyl phthalate, β-sitosterol and daucosterol on K562 cells (K562 cells were treated with dibutyl phthalate, β-sitosterol and daucosterol at various concentrations for 24 h (A), 48 h (B), 72 h (C) and cell viability was determined by MTT assay. Data were shown as mean ± SD (n = 6). #p < 0.05, ##p < 0.01 compared with the control).