ORMDL3剪接异构体的鉴定及在反复喘息患儿中的表达水平分析

Identification of ORMDL3 splice variant and analysis of its expression level in recurrent wheeze children

目的对反复喘息儿童外周血中血清类粘蛋白1样蛋白3(ORMDL3)的剪接异构体ORMDL3-v1进行鉴定并对其表达水平进行分析。方法反复喘息儿童外周血提取RNA,根据ORMDL3-v1的mRNA序列,设计引物行5′-RACE实验,鉴定其是否存在。随后选择48名患有反复喘息的5岁以下儿童,分为喘息A组28例和喘息B组20例;正常对照组选取的是32例正常体检的儿童,检测外周血ORMDL3-v1及其野生型ORMDL3基因的表达水平。同时计算ORMDL3-v1/ORMDL3的比值,并分析各组病例的临床特点。结果5′-RACE实验证实反复喘息儿童外周血中存在ORMDL3-v1的表达。正常对照组ORMDL3-v1及ORMDL3基因表达水平都显著低于喘息A组和B组(P<0.01),喘息B组ORMDL3的表达水平显著低于A组(P<0.01),喘息A组和B组ORMDL3-v1的表达水平在统计学上差异不明显(P>0.05)。喘息A组ORMDL3-v1/ORMDL3的比值与喘息B组无差异(P>0.05),二者均低于正常对照组(P<0.01)。结论ORMDL3-v1及ORMDL3的表达水平与5岁以下儿童的反复喘息密切相关。

Objective To identify the splice variant of orosomucoid 1-like protein 3(ORMDL3)and analyze the expressions of ORMDL3-v1 in recurrent wheeze children.Methods RNA was extracted from the peripheral blood of children with recurrent wheeze.Based on the mRNA sequence of ORMDL3-v1,primers were designed for 5'-RACE experiments to identify its presence.48 children under 5 years old were divided into the atopic wheezing group A(28 cases)and the non-atopic wheezing group B(20 cases).32 cases of healthy children were selected in the normal control group.The expression level of ORMDL3 and its splice variant ORMDL3-v1 mRNA were measured by RT-PCR.The ratio of ORMDL3-v1/ORMDL3 was calculated.The clinical characteristics of each group were analyzed.Results 5′-RACE experiment confirmed the expression of ORMDL3-v1 in the peripheral blood of children with recurrent wheeze.The expression levels of ORMDL3-v1 and ORMDL3 genes in the normal control group were lower than those in the wheezing group A and B(P<0.01).The expression level of ORMDL3 in the wheezing group B was lower than those in the group A(P<0.01).The expression level of ORMDL3-v1 in the wheezing group A and B were not statistically significant(P>0.05).The ratio of ORMDL3-v1/ORMDL3 in the wheezing group A was not different from that in the wheezing group B(P>0.05).The ratio of ORMDL3-v1/ORMDL3 in the normal control group were higher than those in the wheezing group A and the group B(P<0.01).Conclusion The expression level of ORMDL3-v1 and ORMDL3 is closely related to recurrent wheeze children under 5 years old.