Efficacy of Yisui granule(益髓颗粒)on myelodysplastic syndromes in SKM-1 mouse xenograft model through suppressing Wnt/β-catenin signaling pathway

OBJECTIVE:To unmask the underlying mechanisms of Yisui granule(益髓颗粒,YSG)for the treatment of Myelodysplastic syndromes(MDS).METHODS:Our study used an SKM-1 mouse xenograft model of MDS to explore the anti-tumor potential of YSG and its safety,assess its effect on overall survival(OS),and evaluate whether its mechanism is associated with the demethylation of the secreted frizzled related protein 5(s FRP5)gene and suppressing Wnt/β-catenin pathway.Bisulfite amplicon sequencing was applied to detect the level of methylation of the s FRP5 gene;western blotting,immunofluorescence staining,and real-time Polymerase Chain Reaction were performed to detect DNA methyltransferase 1(DNMT1),s FRP5,and other Wnt/β-catenin pathway-related m RNA and protein expression.RESULTS:The results showed that high-dosage YSG exerted an anti-tumor effect similar to that of decitabine,improved OS,and reduced long-term adverse effects in the long term.Mechanically,YSG reduced the expression of DNMT1 methyltransferase,decreased the methylation,and increased the expression of the Wnt/β-catenin pathway antagonist-s FRP5.Furthermore,components of the Wnt/β-catenin pathway,including Wnt3a,β-catenin,c-Myc,and cyclin D1,were down-regulated in response to YSG,suggesting that YSG could treat MDS by demethylating the s FRP5 gene and suppressing the Wnt/β-catenin pathway.CONCLUSIONS:Our findings demonstrated that YSG could be used alone or in combination with decitabine to improve outcomes in the MDS animal model,providing an alternative solution for treating MDS.

Effect of Yisui Shengxue Granule (益髓生血颗粒) on the Oxidative Damage of Erythrocytes from Patients with Hemoglobin H Disease

Objective： To investigate the effect of Yisui Shengxue Granule （益髓生血颗粒, YSSXG）, a complex Chinese medicine, on the oxidative damage of erythrocytes from patients with hemoglobin H （HbH） disease. Methods： Twenty-two patients with HbH disease and 22 healthy volunteers were observed. YSSXG was given to patients with HbH disease for 3 months. Before and after the 3-month treatment, blood parameters [hemoglobin （Hb）, red blood cells （RBCs）, and reticulocyte percent （Ret）] were examined; inclusion bodies in erythrocytes were observed by transmission electron microscopy （TEM）; activities of antioxidant defense enzymes [superoxide dismutase （SOD）, glutathione peroxidase （GSH-Px）, and catalase （Cat）] and erythrocyte membrane malondialdehyde （MDA） concentrations were determined. Results： In patients with HbH disease, measured values of RBC and Hb obtained from the first to the third months after treatment with YSSXG were significantly higher than before treatment （P〈0.01）. Measured values of Ret from the second to the third months aftertreatment were significantly lower than before treatment （P〈0.05 and P〈0.01, respectively）. Prior to treatment with YSSXG, TEM images of RBCs showed the presence of numerous inclusion bodies. After treatment with YSSXG, the amount and volume of inclusion bodies decreased. Treatment with YSSXG also led to a significant increase in SOD activity （P〈0.01）, a decrease in Cat activity （P〈0.01）, and no significant differences in GSH-Px activity （P〉0.05） or MDA concentration （P〉0.05）. However, compared with the healthy counterparts, SOD, GSH-Px, and Cat activities presented at high levels （P〈0.01） both before and after treatment. Conclusions： YSSXG could improve the degree of hemolysis and anemia in patients with HbH disease. The mechanism may be related to its antioxidative effects, which could elevate the activity of total SOD in erythrocytes and efficiently inhibit the oxidative precipitation of β-globin chains.