Positive And Negative Control Roles Of Agaricus Blazei Murrill-Qbsg Aqueous Extract On Human Bone Marrow Cells

Agaricus blazei Murrill-Qbsg is a kind of selenium-rich edible fungus successfully cultivated in China. In order to study its water-soluble components'negative and positive control roles to hematological system cells, we extracted Agaricus blazei Murrill-Qbsg's aqueous extract under 4 different temperature, respectively 4℃, 25℃, 60℃, 100℃, which we used to effect on Promyelocytic leukemia cells(K562)to obtain the best one. In the process of study of Agaricus blazei Murrill-Qbsg aqueous extract inducing apoptosis of K562 cells, MTT, Colony Formation, FACS, AGE, Fluorescence microscope, TEM and Single-cell RT-PCR were used to do the activity detection, biochemical detection, morphology observation and Apoptosis-related gene expression. We used our own lab-made Immunologic Nano-Fluorescence Beads(INFB)and FACS to survey in the couse of study of Agaricus blazei Murrill-Qbsg aqueous extract effecting on human cord blood cells differentiation antigen. In this study we use MTT to identify if our own lab-made Nano-Fluorescence Beads have any toxic effect on Mouse bone marrow cells, K562 cells and Hela cells. And we also try to use our own lab-made Nano-Magnetism Beads to extract mRNA directly from cell extraction buffer.In the study of Agaricus blazei Murrill-Qbsg aqueous extract inducing apoptosis of K562 cells, the results indicated that Agaricus blazei Murrill-Qbsg aqueous extract under 60℃has the best inhibitory action on K562 cells among all. And we also found out that it could strongly inhibit the growth of K562 cells, P<0.01, compared to the control group. And the findings of this part of study showed that there was a positive correlated between treatment hour and inhibition ratio during the treatment hours from 24h to 72h. The results examined by FACS indicated that it appeared a significant subdiploid when K562 cells had been treated for 72h. Further studies involving fluorescence and TEM revealed characteristic apoptotic features like membrane blebbing, karyopyknosis, chromatin condensation, forming crescent-shaped and apoptotic body. Agarose gel electrophoresis of DNA from K562 cells treated with Agaricus blazei Murrill-Qbsg aqueous extract revealed"ladder"pattern. Through single cell RT-PCR, we found out that Agaricus blazei Murrill-Qbsg aqueous extract could increase the expression of p53, Fas, Caspase-3. In the study of Agaricus blazei Murrill-Qbsg aqueous extract effecting on hemopoietic progenitor cells differentiation antigen, FACS analytic results of human umbilical cord blood hemopoietic stem cells treated with Agaricus blazei Murrill-Qbsg aqueous extract indicated that it could promote efficiently the differentiation of mononuclear cells to CFU-E,CFU-GM,B cell,T cell. According to the results of MTT, we found that our own lab-made Nano-Fluorescence Beads had not toxic effect on Mouse bone marrow cells, K562 cells and Hela cells; theβ-actin RT-PCR results showed that our own lab-made Nano-Magnetism Beads could separate mRNA directly from cell extraction buffer, which could be used to gene amplification.Therefore, the active ingredients of Agaricus blazei Murrill-Qbsg aqueous extract under 60℃have positive and negative two different kinds of control roles, inducing apoptosis of K562 cells and promoting the differentiation of mononuclear cells. Our own lab-made Nano-Fluorescence Beads have no significant toxic effect on the growth of cells, which provides the research basis in vivo. Meanwhile, Nano-Magnetism Beads we made could separate mRNA effectively and directly from cell extraction buffer, which provides the conditions for the rapid separation mRNA.