摘要
目的研究Stomatin样蛋白2(STOML2)基因表达在儿童哮喘发病中的作用。方法选择2013年9月-2014年3月在西安市儿童医院(以下简称"我院")呼吸科住院的14例哮喘患儿为研究对象,选择同期在我院进行健康体检的9例正常儿童作为对照。采用RT-q PCR和Western blot方法检测两组儿童外周血单个核细胞(PBMC)中STOML2和磷脂酰肌醇3激酶(PI3K)m RNA和蛋白的表达水平;构建STOML2过表达载体并转染气道平滑肌细胞(ASMC),MTT法检测细胞的生长,观察STOML2过表达对PI3K表达的影响。结果与正常儿童比较,哮喘患儿PBMC中STOML2 m RNA及蛋白水平显著降低,而PI3K水平显著升高,差异均有统计学意义(P〈0.05)。体外实验结果显示,过表达STOML2会抑制气道平滑肌细胞生长,并抑制PI3K的表达。结论 STOML2在哮喘患儿中低表达,过表达STOML2抑制ASMC的生长,其可能是通过抑制PI3K活性实现的。
Objective To investigate the effect of Stomatin-like Protein 2(STOML2) expression on pathogenesis of childhood asthma. Methods Totally 14 children with asthma who were hospitalized in the Department of Respiration,Xi'an Children's Hospital("our hospital" for short) were recruited from September 2013 to March 2014, and 9 healthy children who received physical check-ups in our hospital at the same period were taken as controls. The m RNA and protein levels of STOML2 and phosphatidylinositol 3 kinase(PI3K) in peripheral blood mononuclear cell(PBMC) of children in two groups were detected by RT-q PCR and Western blot. In addition, STOML2 over-expressed vector was synthesized and transfected to airway smooth muscle cells(ASMC). The cell viability was detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, and the effect of STOML2 over-expression on the level of PI3 K was observed. Results The m RNA and protein levels of STOML2 in PBMC of asthmatic children were significantly decreased when compared with those in normal children, while the PI3 K level was markedly increased, the differences were statistically significant(P〈0.05). In vitro assay showed that over-expression of STOML2 inhibited the growth of ASMC and the expression of PI3 K. Conclusion STOML2 is decreased in asthmatic children and over-expression of STOML2 inhibited the growth of ASMC, which is accompanied by inhibiting PI3 K pathway.
出处
《中国医药导报》
CAS
2016年第17期20-24,共5页
China Medical Herald