Effects Of Herba Saginae Japonicae On Proliferate Inhibition And Apoptosis Induction In HL-60Cell Strain

Objective To investigate the effects of Herba Saginae Japonicae（HSJ）on proliferate inhibition and apoptosis induction in HL-60cells, and researchthe mechanism of the induction. Methods HL-60cells were treated by variousdosages of HSJ. Through detecting the proliferate-inhibition ratio by methyl thiazolyltetrazolium(MTT) colorimetry, effect on growth curve, and the morphology change ofHL-60after affected by HSJ, to evaluate the proliferate-inhibition capability; Theapoptosis induction of HSJ on HL-60cells were detected by Annexin V-FITC/PIdouble staining and Hoechst33258fluorescent dye; the effect of HSJ onmitochondrial membrane potential of HL-60cells was examined by JC-1fluorescentdye; the expression level of Bcl-2, Bax, caspase-3and caspase-9were calculated bywestern blot. Results (1) The MTT results showed that HSJ could remarkably inhibitthe HL-60cells proliferation, and the IC50value at48h of treatment, as about20.17μg/ml.(2) HL-60cells after treatment with HSJ4μg/ml,20μg/ml,100μg/ml for48h, the prophase apoptotic ratios by Annexin V-FITC/PI double staining wereseparately （6.42±0.04）%,（9.76±0.06）%,（11.23±0.02）%, significantly differentcomparing with the control (P﹤0.05).(3) HL-60cells after treatment with HSJ4μg/ml,20μg/ml,100μg/ml for48h, the apoptotic ratios by Hoechst33258fluorescent dye were separately（10.06±0.13）%,（29.34±0.72）%,（33.09±0.75）%, significantly different comparing with the control (P﹤0.05); HL-60cells treatedwith HSJ, having condensed cell nucleus, assembled chromatin, disintegrated nucleusbody and the majority of the cells appeared to be apoptotic conformation.(4) HL-60cells after treatment with HSJ4μg/ml,20μg/ml,100μg/ml for48h，mitochondrialmembrane potential by JC-1fluorescent dye were separately（15.6±4.45）%、（30.2±6.49）%、（57.8±9.68）%.(5) Western blot results indicatd that HSJ couldinhibit the expression levels of Bcl-2and up-regulate the expression of Bax, caspase-3 and caspase-9, significantly different comparing with the control (P﹤0.05).Conclusion HSJ may inhibit proliferation of HL-60cells and induce HL-60cells toundergo apoptosis; HSJ induces apoptosis of HL-60cells by down regulating theexpression levels of Bcl-2and up regulating the expression levels of Bax, caspase-3and caspase-9.