Effect of resveratrol on mouse model with blood deficiency syndrom and mechanism in inducing erythroid differentiation of K562 cells
Objective To determine the effect of resveratrol(RES)on the hematopoietic function in mice with blood deficiency syndrome induced by cyclophosphamide,and the mechanism of RES in inducing erythroid differentiation of K562 cells.Methods Cyclophosphamide was used to induce the mouse model of blood deficiency syndrome.After the modeling,BALB/c mice were randomly divided into a control group,a model group,and low,medium,and high dose RES groups(50,100 and 200 mg·kg-1).The mice were given the drug continuously for 10 days.On the 11th day,the peripheral blood cells of mice were analyzed with a blood cell analyzer.The concentration of erythropoietin(EPO)and interleukin-6(IL-6)in the plasma of mice were determined by enzyme linked immunosorbent assay.The apoptosis,cell cycle distribution and the expression of erythroid differentiation marker proteins CD71 and CD235a of K562 cells induced by different concentrations of RES were measured by flow cytometry.The mRNA expressions of CD235a,CD71,GATA1,γ-globin,HBA1,HBB and ZFPM1 were detected by RT-qPCR,and the expressions of JNK1/2 and ERK1/2 were detected by Western blot.Results Compared with the model group,the white blood cells,red blood cells,platelets,reticulocyte and hemoglobin in the blood of mice in RES groups were all increased(P＜0.05).The expressions of EPO and IL-6 in the RES groups were significantly increased(P＜0.05).Flow flow cytometry showed that compared with the control group,RES inhibited the proliferation via S phase block and induced the apoptosis of K562 cells,and greatly increased the positive expression rate of erythroid surface marker proteins CD235a and CD71 in a dose-dependent manner(P＜0.05).RT-qPCR showed that the expressions of erythroid differentiation related genes such as CD235a,CD71,GATA1,γ-globin,HBA1,and HBB in K562 cells were significantly increased after the treatment with different concentrations of RES,also in a dose-dependent manner(P＜0.05).Western blot showed that RES up-regulated the phosphorylation levels of ERK1/2 and JNK1/2/3 proteins.Conclusion By regulating JNK pathway and ERK pathway,RES can increase the positive expression of CD71 and CD235a cell surface antigens,increase the expression of key genes of erythroid differentiation,induce K562 cells to differentiate into erythroid cells,and ameliorate the symptoms of blood deficiency syndrome caused by cyclophosphamide in mice.The study lays a foundation for the mechanism of RES promoting hematopoiesis.
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