Iftikhar Fizza, Ali Hamad, Musharraf Syed Ghulam
Dr Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi Karachi-75270 Pakistan
H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi Karachi-75270 Pakistan.
RSC Adv. 2019 Jun 4;9(31):17551-17559. doi: 10.1039/c9ra01744e.
Pharmacologically mediated reactivation of γ-globin gene with an increase in fetal hemoglobin production, is a cost effective experimental therapeutic intervention for the management of β-hemoglobinopathies. Investigation of new pharmacological agents as HbF inducers from natural resources is desirable to develop safe and effective HbF inducers. We evaluated selected cinchona alkaloids (cinchonidine and quinidine) for their potential of erythroid differentiation and augmentation of fetal hemoglobin production. K562 cells were used as experimental model. Erythroid differentiation of K562 cells was studied using a benzidine assay, and total hemoglobin was estimated through a calorimetric method. Whereas, quantitative real-time PCR (qRT-PCR) was used to analyse γ-globin gene expression, and flow cytometry and immunofluorescence microscopy for evaluating HbF production. Cinchona alkaloids showed dose dependent erythroid differentiation, time driven cellular proliferation, with kinetics of hemoglobin accumulation in K562 cells. The findings of qRT-PCR showed an increase in expression of γ-globin mRNA content (3.17-fold in cinchonidine and 2.03-fold increase in quinidine treated K562 cells), accompanied by an increase in fetal hemoglobin production. Altogether, this study demonstrates that cinchona alkaloids can be used as therapeutic agents in treating β-thalassemia after further biological investigation.
通过药理作用介导γ-珠蛋白基因重新激活并增加胎儿血红蛋白的产生，是一种治疗β-地中海贫血的经济有效的实验性治疗干预措施。研究从自然资源中筛选新型作为胎儿血红蛋白诱导剂的药理活性剂，对于开发安全有效的胎儿血红蛋白诱导剂具有重要意义。我们评估了选定的金鸡纳生物碱（辛可尼定和奎尼丁）在诱导红细胞分化和增加胎儿血红蛋白产生方面的潜力。以K562细胞作为实验模型。使用联苯胺试验研究K562细胞的红细胞分化，并通过比色法估算总血红蛋白含量。同时，采用定量实时聚合酶链反应（qRT-PCR）分析γ-珠蛋白基因表达，并通过流式细胞术和免疫荧光显微镜评估胎儿血红蛋白的产生。金鸡纳生物碱在K562细胞中呈现出剂量依赖性的红细胞分化、时间驱动的细胞增殖以及血红蛋白积累的动力学特征。qRT-PCR结果显示，γ-珠蛋白mRNA含量表达增加（辛可尼定处理的K562细胞增加3.17倍，奎尼丁处理的K562细胞增加2.03倍），同时胎儿血红蛋白的产生也增加。总之，本研究表明，经过进一步生物学研究后，金鸡纳生物碱可作为治疗β-地中海贫血的治疗药物。
