川芎嗪对支气管上皮细胞ORMDL3表达影响

EFFECT OF LIGUSTRAZINE ON THE EXPRESSION OF OROSOMUCOID 1-LIKE PROTEIN 3 IN BRONCHIAL EPITHELIAL CELLS

目的探讨川芎嗪对哮喘易感基因血清类黏蛋白1样蛋白3(ORMDL3)表达的影响。方法应用IL-4+IL-13刺激人支气管上皮(HBE)细胞ORMDL3过表达,以不同浓度的川芎嗪(200、300、400、500、600、700、800μg/L)干预72 h,选取干预作用最明显的川芎嗪浓度。以IL-4+IL-13培养的HBE细胞作为模型组,以干预作用最明显的川芎嗪浓度干预IL-4+IL-13培养HBE细胞作为治疗组,PBS培养HBE细胞作为空白组。应用RT-PCR方法分别检测3组ORMDL3 mRNA表达,Western blot方法、免疫组化法检测ORMDL3蛋白表达。结果400μg/L浓度的川芎嗪干预作用最明显。治疗组、空白组、模型组ORMDL3 mRNA表达比较差异有显著性(F=176.45,P<0.01),治疗组表达最低。模型组、治疗组ORMDL3蛋白表达较空白组升高,治疗组较模型组降低,差异有统计学意义(F=23.48、91.56,P<0.01)。结论川芎嗪可通过下调HBE细胞ORMDL3基因表达,改善哮喘病人气道重塑。

Objective To investigate the effect of ligustrazine on the expression of the asthma susceptibility gene orosomucoid 1-like protein 3(ORMDL3)in serum.Methods Human bronchial epithelial cells(HBECs)were stimulated by interleukin-4(IL-4)and interleukin-13(IL-13)to induce the overexpression of ORMDL3,and ligustrazine at different concentrations(200,300,400,500,600,700,and 800μg/L)was used for intervention for 72 h to screen out the concentration of ligustrazine with the most significant intervention effect.HBECs cultured with IL-4+IL-13 were selected as model group;HBECs cultured with ligustrazine at the concentration with the most significant intervention effect were selected as treatment group;HBECs cultured with phosphate-buffered saline were selected as blank group.RT-PCR was used to measure the mRNA expression of ORMDL3,and Western blot and immunohistochemistry were used to measure the protein expression of ORMDL3.Results Ligustrazine at the concentration of 400μg/L had the most significant intervention effect.There was a significant difference in the mRNA expression of ORMDL3 between the treatment group,the blank group,and the model group(F=176.45,P<0.01),and the treatment group had the lowest expression of ORMDL3.The model group and the treatment group had significantly higher protein expression of ORMDL3 than the blank group,and the treatment group had significantly lower expression than the model group(F=23.48,91.56;P<0.01).Conclusion Ligustrazine can improve airway remodeling in asthmatic patients by downregulating ORMDL3 gene expression in HBECs.