Study Of Loranthaceae Effective Parts Of Anti-leukemia In Vitro Screening And Induction Leukemia Cell Apoptosis

Objective To observe the influence of leukemic K562, HL-60 cell proliferation and cell morphology arising from the different extracts of Loranthaceae in vitro. The initial screening effective part of Loranthaceae anti-leukemia;Study of apoptosis of leukemia cells induced by the different aspects of Loranthaceae extracts and its mechanism, from apoptosis and mitochondrial membrane potential; Then using the method of serum pharmacology to observe Loranthaceae containing serum on effect of cell proliferation and apoptosis of leukemia K562;To provide the basis of preliminary laboratory for further study of the pharmacological mechanism of Loranthaceae and its components anti-leukemia and clinical applications.Methods Using MTT assay to detect the cell proliferation inhibiting action on K562, HL-60 cells induced by the different extracts of Loranthaceae after 24h,48h,72h, to obtain inhibition rates and calculate the IC50, meanwhile cell morphology was observed by inverted microscope to initially select effective parts of Loranthaceae anti-leukemia, then choose the effective parts of the Loranthaceae:water extract, ethyl acetate parts, butanol parts, chloroform parts and ether parts as the experimental group (according to MTT results, to set up high, middle, low concentrations), Ara-C as a positive control,1% DMSO in serum-free culture medium as blank control, respectively act on K562, HL-60 cells in 48h, Then the changes of apoptosis and mitochondrial membrane potential are measured and analyzed by flow cytometry(Apoptosis is tested by Annexin V/PI double staining and mitochondrial membrane potential changes were detected with Rhodamine 123 staining). Combining with the method of serum pharmacology,MTT method and flow cytometry are adopted to detect the effect of cell proliferation and apoptosis of leukemia K562 produced by Loranthaceae containing serum. Finally, the test data is inputted SPSS17.0 to establish a database for statistical analysis. Result By MTT anti-tumor experiments in vitro, results showed that the Loranthaceae has a certain anti-leukemia activity, namely Loranthaceae is more sensitive to the K562. Loranthaceae anti-leukemia cell proliferation, the role of the longer, the higher the concentration, the stronger the inhibition of cell, showing obvious time-dose dependent manner;the same time, the state of cell growth also have significant impact, Cell density and cell morphology show the changes of apoptotic cells. MTT results showed that the differences between the role of different parts of Loranthaceae, including ethyl acetate parts, chloroform parts and butanol parts showed stronger activity area;aqueous extract, ether parts is less important, petroleum ether parts minimal impact on the cells. Flow cytometry showed that Loranthaceae effective parts act on the K562, HL-60 cells, induced early apoptosis and decreased cell mitochondrial membrane potential, experimental group and control group P<0.05, significant difference. The study of serum containing showed that Loranthaceae containing serum can inhibit the proliferation of leukemia K562 cells and induce early apoptosis, and show a dose-dependent manner, compared with the blank rabbit serum group P<0.05, significant difference.Conclusion Research suggests that the Loranthaceae have anti-leukemia activity, by inhibiting proliferation of the leukemia cells, induce apoptosis, The apoptosis mechanism may include reducing the mitochondrial membrane potential, and thus the changes of intracellular environmental may cause leukemia cells death.