Sahar Namoodn, Bibi Shakila, Masood Nosheen, Faryal Rani
Department of Biosciences, COMSATS Institute of Information Technology, Islamabad, Pakistan.
Department of Environmental Sciences, Fatima Jinnah Women University, Rawalpindi, Pakistan.
Mol Biol Res Commun. 2019 Jun;8(2):69-77. doi: 10.22099/mbrc.2019.33040.1394.
Asthma is a disease marked by inflammation of airways with an increasing incidence rate worldwide especially among Asian population. Spleen tyrosine kinase (Syk) is known to be involved in regulation of such inflammation response and thereby rendering its inevitable importance among asthma patients. DNA extraction followed by PCR and sequencing was performed for genomic analysis, mRNA analysis was done by RT PCR whereas Western blot and ELISA was used for protein study. Image J and UNAFOLD were also used for Bioinformatics analysis.The mean age of patients and controls were 31.1±9.3 and 30.4±6.1 years respectively. Results of sequencing showed nonsense exonic mutations in exon 3 at g.25710G>A and g.25722GA positions. Substitution mutations in introns were also found at g.25827G>A (intron 3), g.63425C>T (intron 8) and g.63445T>G (intron 8). Significantly increased levels of IgE and significantly decreased expression of Syk at transcriptional level was found in patients compared to controls. The western blot results of asthmatic samples and healthy controls revealed that Syk has comparatively low expression in diseased individual's PBMCs. has been found to be altered in DNA, mRNA and protein expression in asthma patients among Pakistani population therefore patients should be treated according to their Syk status for more effective recovery.
哮喘是一种以气道炎症为特征的疾病，在全球范围内发病率不断上升，尤其是在亚洲人群中。已知脾酪氨酸激酶（Syk）参与这种炎症反应的调节，因此在哮喘患者中具有不可忽视的重要性。进行DNA提取，随后进行PCR和测序以进行基因组分析，通过RT PCR进行mRNA分析，而蛋白质研究则使用蛋白质免疫印迹法和酶联免疫吸附测定法。Image J和UNAFOLD也用于生物信息学分析。患者和对照组的平均年龄分别为31.1±9.3岁和30.4±6.1岁。测序结果显示，外显子3在g.25710G>A和g.25722GA位置存在无义外显子突变。在内含子中也发现了替代突变，分别位于g.25827G>A（内含子3）、g.63425C>T（内含子8）和g.63445T>G（内含子8）。与对照组相比，患者的IgE水平显著升高，Syk在转录水平的表达显著降低。哮喘样本和健康对照的蛋白质免疫印迹结果显示，Syk在患病个体的外周血单核细胞中的表达相对较低。在巴基斯坦人群的哮喘患者中，已发现DNA、mRNA和蛋白质表达发生改变，因此应根据患者的Syk状态进行治疗，以实现更有效的康复。
