固本防哮饮对哮喘易感基因ORMDL3以及信号转导及转录激活因子1的影响

发布时间：2018-05-29 18:37

本文选题：固本防哮饮 + 哮喘　；参考：《南京中医药大学》2016年硕士论文

【摘要】：目的：探讨中药复方固本防哮饮对哮喘易感基因血清类粘蛋白1样蛋白3(orosomucoid 1-like protein 3,ORMDL3)以及信号转导及转录激活因子1(signal transducer and activator of transcription 1, STAT1)的调控作用,进一步揭示固本防哮饮防治哮喘的机制。方法：体外培养人支气管上皮细胞(human bronchial epithelial cells,16HBE),分为正常组、模型组、固本防哮饮组及孟鲁司特钠组,利用白介素-4(interleukin-4, IL-4)诱导16HBE构建哮喘细胞模型,并分别予空白血清及含药血清进行干预；运用实时荧光定量-PCR (Real-time PCR)、免疫印迹法(Western blotting, WB)、免疫荧光法(immunofluorescence, IF)从基因、蛋白水平检测16HBE细胞ORMDL3、STAT1的表达及蛋白亚细胞定位。结果：ORMDL3蛋白主要表达于16HBE细胞胞质中表达,STAT1蛋白可表达于16HBE细胞胞质及胞核内；WB结果显示,与正常组比较,模型组ORMDL3、STAT1蛋白表达显著增加(P0.05,P0.01)；与模型组比较,固本防哮饮组ORMDL3及STAT1蛋白表达均显著下降(P0.05)；PCR结果显示,与正常组比较,模型组ORMDL3、STAT1 mRNA含量升高,但二者无显著性差异(P0.05),固本防哮饮组较模型组细胞中ORMDL3、STAT1 mRNA表达水平下降,无显著性差异(P0.05)；相关性分析显示,ORMDL3 mRNA及STAT1 mRNA表达存在正相关(Pearson=0.858, P0.01)。结论：体外实验表明,固本防哮饮含药血清对哮喘易感基因ORMDL3及转录因子STAT1的表达有一定的调控作用,STAT1参与固本防哮饮对哮喘易感基因ORMDL3的调控可能是固本防哮饮防治哮喘作用的内在机制。
[Abstract]:Objective: to investigate the regulatory effects of Gubenfangxiao Yin (GBFY) on serum mucin-like protein (3(orosomucoid 1-like protein _ 3) and signal transduction and transcription activator (1(signal transducer and activator of transcription _ 1, STAT _ 1) of susceptibility gene to asthma. Further reveal the mechanism of Guben Fangxiao Yin in prevention and treatment of asthma. Methods: human bronchial epithelial cells were cultured in vitro and divided into normal group, model group, Gubenfangxiao decoction group and montelukast sodium group. The asthmatic cell model was induced by interleukin-4 (IL-4). The expression and subcellular localization of ORMDL3T1-STAT1 in 16HBE cells were detected by real-time fluorescence quantification-PCR real-time PCRG, Western blotting, WBX, and immunofluorescence fluorescence assay (IFFI) respectively in blank serum and drug-containing serum, and the expression of ORMDL3T1-STAT1 and protein subcellular localization in 16HBE cells were detected by Western blotting, Western blotting, immunoblorescence (WBX) and immunofluorescence assay. Results the expression of statin T1 protein in the cytoplasm of 16HBE cells was mainly expressed in the 16HBE cells. The results showed that the expression of STAT1 protein in the model group was significantly higher than that in the normal group, and that in the model group was significantly higher than that in the model group, while in the model group, the expression of STAT1 protein in the cytoplasm and nucleus of the 16HBE cells was significantly higher than that in the control group. Compared with the normal group, the expression of ORMDL3 and STAT1 protein in the model group increased, but there was no significant difference between the two groups (P 0.05). The expression of ORMDL3T1STAT1 mRNA in the Guben Fangxiao group was lower than that in the model group. There was no significant difference in the expression of ORMDL3 mRNA and STAT1 mRNA, but there was a positive correlation between the expression of ORMDL3 mRNA and STAT1 mRNA. Conclusion: in vitro experiments show that, The expression of susceptibility gene ORMDL3 and transcription factor STAT1 in the serum of Guben Fangxiao Yin was regulated to some extent. STAT1 involved in the regulation of Guben Fangxiao Yin on the asthma susceptibility gene ORMDL3 may be the internal mechanism of Guben Fangxiao Yin to prevent and cure asthma.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285

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