摘要
目的 探讨甲异靛体外对外周血培养红系细胞的珠蛋白mRNA相对含量的影响 ,为临床治疗 β地中海贫血提供理论依据。方法 采用二步法液体培养外周血红系细胞和逆转录 聚合酶链反应 (RT PCR)等方法 ,研究了 6例正常人和 11例 β地中海贫血患儿外周血体外培养红系细胞在不同浓度甲异靛作用下 ,以α珠蛋白mRNA为内对照 ,测定γ和 β珠蛋白mRNA的相对含量。 结果( 1)甲异靛对所有外周血体外培养红系细胞的β/αmRNA比值变化均无影响。 ( 2 )经不同浓度甲异靛诱导后正常人外周血体外培养的红系细胞的γ/αmRNA升高 0 31～ 0 4 5倍 ,非α/αmRNA升高0 2 1～ 0 32倍 ,三种不同浓度之间差异无显著性 ;β地贫患儿γ/αmRNA升高 0 33～ 1 17倍 ,非α/αmRNA升高 0 2 5～ 0 89倍 ,2 5 μmol/L和 5 μmol/L两种浓度之间差异无显著性 ,10 μmol/L浓度甲异靛对γ/αmRNA和非α/αmRNA的比值变化比上述两种浓度显著增高。( 3) 10 μmol/L甲异靛对 β地贫患儿外周血体外培养红系细胞的γ/αmRNA和非α/αmRNA的比值的提高显著高于正常人 ,其他两种浓度下差异无显著性。结论 甲异靛在体外培养的外周血红系细胞中提高γ/αmRNA和非α/αmRNA的比率 ,对 β地中海贫血的治疗具有潜在应用价值。
Objective The pathophysiology of β thalassemia is the imbalance of the α and non α globin chain which leads to a series of clinical symptoms of hemolytic anemia Scientists continuously try to explore gene activated drugs to increase the level of non α globin chain or decrease the level of α globin chain in the treatment of β thalassemia To probe into the effects on globin gene expression of meisoindigo(Me) in cultured erythroid cells derived from peripheral blood, so as to provide the theoretical basis for applying Me in the treatment of β thalassemia Methods By using the two step liquid culture of erythroid progenitor cells and reverse transcription polymerase chain reaction (RT PCR), and by using α mRNA as an inner control, the level of γ mRNA and β mRNA in cultured erythroid cells derived from peripheral blood of 11 patients with severe β thalassemia and 6 normal volunteers were measured under the effect of different concentration (2 5 μmol/L, 5 μmol/L and 10 μmol/L) of Me Results (1) No statistic significance was found in the ratio of β/α mRNA by Me in cultured cells from both normal individuals and β thalassemia (2) Me can significantly increase the ratio of γ/α mRNA and (β+γ)/α mRNA(that is non α/α mRNA) in cultured cells from normal individuals and β thalassemia The ratio of γ/α mRNA was increased 0 31～0 45 times and the ratio of non α mRNA/α mRNA increased 0 21～0 32 times in Me induced cells from normal individuals No significant result was observed among the different concentrations of Me (2 5 μmol/L, 5 μmol/L and 10 μmol/L) in normal individuals With the increasing of Me concentrations, the ratios of γ/α mRNA and α/α mRNA were increased in cultured cells from β thalassemia The ratio of γ/α mRNA was increased 0 33～1 17 times and the ratio of non α/α mRNA increased 0 25～0 89 times in Me induced cells from β thalassemia There was no significant difference between the concentrations of 2 5 μmol/L and 5 μmol/L concentration in β thalassemia However, there was significant difference between the concentrations of 10 μmol/L and the concentrations of 2 5 μmol/L and 5 μmol/L in β thalassemia (3) The increase of the ratio of γ/α mRNA and non α/α mRNA in β thalassemia was higher than that in normal individual with induction by Me with a higher concentration (10 μmol/L) Conclusion Me can raise the ratio of γ/α mRNA and non α/α mRNA in cultured erythroid cells derived from peripheral blood of both normal individual and β thalassemia in the level of transcription, which can improve the imbalance of the α and non α globin chain So Me has a latent value in the therapy of β thalassemia
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2004年第3期184-187,共4页
Chinese Journal of Pediatrics
