Objective: To examine the clinical effects of Yisui Shengxue Granules(益髓生血颗粒) in the treatment of β-thalassemia and explore its mechanism on DNA methylation levels. Methods: A randomized placebo-controlled double-blinded trial was conducted. Forty patients with β-thalassemia were recruited and distributed randomly by envelope method into an experimental group and a control group, 20 patients in each group. The patients were given Yisui Shengxue Granules in the experimental group and placebo in the control group(12 g/bag, 3 times a day) during a 3-month intervention. Before and after 1, 2, and 3 months of treatment, peripheral intravenous blood was sampled, and blood parameters such as hemoglobin(Hb), red blood cells(RBCs), reticulocytes(Ret), and fetal hemoglobin(HbF) were analyzed. Mononuclear cells from 5 patients, who showed an obvious treatment effect, were isolated by density gradient centrifugation. DNA methylation was analyzed using an Affymetrix USA GeneChip Human Promoter 1.0 Array and Input-promoter 1.0. Results: Compared with pre-treatment, there was an obvious increase in Hb and RBCs counts after 1, 2, and 3 months in the experiment group(P<0.01 or P<0.05). Meanwhile, HbF increased from the 2 nd to the 3 rd month(P<0.05). In the control group, Hb and RBCs showed no obvioas change. After 3-month treatment, DNA methylation results from 5 patients revealed that there were 24 hypomethylated genes and 3,685 hypermethylated genes compared with pre-treatment. Genes of insulin-like growth factor 1 receptor(IGF1 R) and Janus kinase 3(JAK3) revealed the most relations with other genes(degree: 21) and genes of 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2(PLCG2) and mitogen-activated protein kinase 10(MAPK10) showed a stronger intermediary role(betweenness centrality=0.04). Conclusions: JAK3 and MAPK10 are two key genes in bone marrow and the lymphatic system, and JAK3 is likely to be related to hematopoietic cytokines in the process of early hematopoiesis.(Registration No. NCT
目的:观察益髓生血颗粒对再生障碍性贫血(Aplastic Anemia,AA)大鼠CD4^+CD25^+调节性T细胞(Regulatory T Cell,Treg细胞)的影响及其治疗AA的作用机制。方法:取雄性SD大鼠按体重随机分组。其中模型组连续7周隔天皮下注射苯(1 m L?kg^(-1))、取材10天前开始腹腔注射环磷酰胺(25 m L?kg^(-1)),连续3天,第4周将模型组大鼠按体重随机分为模型组、司坦唑醇组、益髓生血颗粒组,灌胃给药。正常组及模型对照组给予等体积生理盐水。实验结束时,检测外周血WBC、RBC、HGB、PLT;制备血涂片、骨髓涂片;免疫组织化学法检测脾组织Treg细胞Foxp3蛋白表达;RT-PCR法检测骨髓组织Foxp3 mRNA的表达。结果:与正常组比较,模型组WBC、RBC、HGB、PLT均显著减少(P<0.01),血涂片示血细胞通透性差、白细胞减少、退化细胞增多,骨髓涂片示脂肪滴明显增加、造血细胞均明显降低、非造血细胞增多;经益髓生血颗粒组治疗后WBC、RBC、HGB、PLT均显著增加(P<0.01),血涂片及骨髓涂片显示细胞通透性好转、形态趋于正常、脂肪滴明显减少,造血细胞均明显增加,脾组织Foxp3蛋白及骨髓组织Foxp3 mRNA表达明显升高(P<0.01)。结论:益髓生血颗粒可上调Foxp3的基因及蛋白表达,调控AA免疫功能,进而改善AA免疫环境,促进骨髓造血功能的恢复,发挥治疗AA的重要作用。
