基于骨髓微环境介导PI3K/Akt通路研究复方君子汤治疗急性髓系白血病机制OA

Objective:To explore the mechanism of action of the compound Fufang Junzi Decoction(FFJZ)in treating acute myeloid leukemia(AML)through the bone marrow microenvironment mediated PI3K/Akt signaling pathway.Methods:A co-culture model of AML cells(K562/VCR)and human bone marrow stromal cells was established to simulate the survival state of leukemia cells in the bone marrow microenvironment.The control group,FFJZ group,and combination group(FFJZ+cytarabine)were set up.Cell morphology changes were observed under a microscope,cell survival rate and proliferation inhibition rate were detected by CCK-8 assay,apoptosis rate was analyzed by flow cytom-etry,and the phosphorylation levels of PI3K,Akt,eIF4B,and the expression of Bcl-2 and Bax proteins were verified by Western blot.The experiments were conducted under two conditions:separate culture and adherent co-culture.Results:①Cell morphology:Compared with the control group,the number of cells in the FFJZ group was significantly reduced(50%),with some cells showing volume shrinkage and irregular morphology;the cell number in the combination group was the most severely reduced(75%),with most cells shrinking in volume and many cell fragments floating in the cell culture medium.②Survival and proliferation inhibition:both the FFJZ group and the combination group significantly reduced the survival rate of K562/VCR cells,and the effect of the combination group was better than that of the single drug group(F=6.808,P<0.001).The survival rate of cells in the co-culture system was higher than that in the sep-arate culture group(F=106.968,P<0.001),and the proliferation inhibition rate was lower than that in the separate culture group(F=103.228,P<0.001).③Apoptosis regulation:in group A,compared with the control group(8.15%),the apoptosis rates in the FFJZ group(20.72%)and the combination group(38.95%)were significantly increased,with the combination group being particularly obvious(P<0.05).In group B,the apoptosis results of each group were consistent with those in group A,and the apoptosis rate of each group was lower than that in group A(F=32.67,P<0.05).4.Signal pathway regulation:in co-cultured cells,the phosphorylation levels of PI3K,Akt,eIF4B,and the expression of Bcl-2 protein were downregulated in the FFJZ group and the combination group(P<0.05),while the expression of Bax protein was upregulated(P<0.05).Conclusion:The combination of FFJZ with cytarabine can enhance the pro-apoptotic effect on AML cells.The mechanism may be related to the bone marrow microenviron-ment-mediated PI3K/Akt pathway,where FFJZ inhibits the activation of the PI3K/Akt pathway,downregulates the phosphorylation of eIF4B,upregulates the pro-apoptotic gene BAX,and downregulates the expression of the anti-apop-totic gene BCL-2,thereby accelerating the apoptosis of leukemia cells and achieving the therapeutic effect on AML.This may provide a reference for AML targeted therapy.