补肾益髓法治疗β-珠蛋白生成障碍性贫血患儿的效果及对抗氧化基因表达的影响

Effect of Busheng Yisui Method on children with β- Thalassemia and antioxidant gene expression variation

目的探讨补肾益髓法治疗β-珠蛋白生成障碍性贫血(β-地中海贫血)的疗效及作用机制。方法观察54例β-珠蛋白生成障碍性贫血患儿补肾益髓法治疗前后血红蛋白(Hb)、红细胞(RBC)、网织红细胞计数(Ret)、胎儿血红蛋白(Hb F)的改变;通过实时定量PCR方法研究患者骨髓有核细胞中珠蛋白编码基因表达量变化以及抗氧化基因的响应情况。结果补肾益髓法治疗后,重型和中间型患者各项血液学参数均有提高,与治疗前比较差异有显著性。重型β-地中海贫血患儿组的Hb、RBC、Ret水平均显著上升(P<0.05),而中间型β-地中海贫血患儿组仅有Hb、RBC水平显著上升(P<0.05)。珠蛋白mRNA变化比值,即γ/(β+γ)mRNA比值在两组患者中均上升,且在重型β-地中海贫血患儿组显著上升(P<0.05)。治疗后发现抗氧化基因的响应,重型β-地中海贫血患儿组骨髓有核细胞超氧化物歧化酶(SOD1),谷氨酰半胱氨酸连接酶催化亚单位(GCLC),硫氧还蛋白(TRX)基因表达均显著高于治疗前(P<0.05);中间型β-地中海贫血患儿组骨髓有核细胞GCLC、TRX基因表达也显著高于治疗前(P<0.05)。治疗前后BTB-CNC异体同源体1(BACH-1)基因表达无显著差异。结论补肾益髓法可以改善β-珠蛋白生成障碍性贫血患儿的临床症状,升高患儿γ珠蛋白基因的表达,有效激活患儿体内抗氧化基因的响应,这可能是补肾益髓法治疗本病的分子机制之一。

Objective To investigate the mechanism of Busheng Yisui Method（ BYM） on Children with β- Thalassemia. Methods The changes of hemoglobin（ Hb）,red blood cells（ RBC）,reticulocyte（ Ret） and fetal hemoglobin（ Hb F） of 54 children with β- Thalassemia were measured before and after Busheng Yisui treatment. The effect of BYM on Globin gene expression and antioxidant gene responses in bone marrow mononuclear cells was determined by real time PCR technique. Results After treatment,hematological parameters were improved both in the major β- Thalassemia group and intermediate β- Thalassemia group,with statistically significant when compared with pre- treatment situation.In the major β- Thalassemia group,levels of Hb,RBC,Ret were significantly increased（ P〈0. 05）,while levels of Hb,RBC were significantly increased in the intermediate β- Thalassemia group（ P〈0. 05）. Ratio of globin mRNA,which was the γmRNA /（ β ＋ γ） mRNA ratio,were rising in both groups of patients,and the change was significant in the major β- Thalassemia group（ P〈0. 05）. After treatment,antioxidant gene response was found. Levels of SOD1,GCLC,TRX gene expression in bone marrow mononuclear cells were significantly higher than pre-treatment in the major β- Thalassemia group（ P〈0. 05）. Levels of GCLC,TRX gene expression in bone marrow mononuclear cells were significantly higher than pre- treatment in the intermediate β- Thalassemia group（ P〈0. 05）. Gene expression level of BACH1 did not differ significantly before and after treatment. Conclusion Busheng Yisui method could improve the clinical symptoms of children with β- Thalassemia. One of the mechanisms may be its effect on increasing the γ- globin gene expression and stimulating the responses of antioxidant genes.