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目的观察FIZZ1在小鼠哮喘模型气道上皮间质转化(EMT)中的作用,探讨FIZZ1引起哮喘早期气道重塑的机制。方法随机将BALB/c小鼠分为哮喘模型组(OVA组)和正常对照组。采用免疫组织化学法测定小鼠气道上皮中FIZZ1蛋白的表达,采用实时定量-聚合酶链反应法检测小鼠肺组织中FIZZ1mRNA、Ⅰ型胶原、纤维连接蛋白1、E钙粘蛋白的表达,采用小鼠肺上皮细胞(MLE-12)体外原代培养,FIZZ1重组蛋白及FIZZ1-shRNA干预,免疫印迹法检测E钙粘蛋白、Ⅰ型胶原和纤维连接蛋白1的表达。结果与对照组相比,OVA组小鼠气道上皮中FIZZ1、FIZZ1mRNA表达显著增强,Ⅰ型胶原和纤维连接蛋白1显著增高,E钙粘蛋白明显降低,并且分别与FIZZ1呈正相关和负相关;体外原代培养MLE-12,FIZZ1重组蛋白干预,Ⅰ型胶原和纤维连接蛋白1增高,E钙粘蛋白显著降低;FIZZ1-shRNA干预,蛋白表达正常。结论FIZZ1作为EMT的一个潜在诱导剂,诱导E钙粘蛋白表达减少,而Ⅰ型胶原和纤维连接蛋白1表达增加,从而诱导哮喘早期气道重塑。
Objective To investigate the role of FIZZ1 in airway epithelial-to-mesenchymal transition (EMT) in mouse asthma model and to explore the mechanism of FIZZ1-induced early airway remodeling in asthma. Methods BALB / c mice were randomly divided into asthma model group (OVA group) and normal control group. The expression of FIZZ1 protein in airway epithelium of mice was detected by immunohistochemical method. The expression of FIZZ1 mRNA, type Ⅰ collagen, fibronectin 1 and E-cadherin in lung tissue were detected by real-time quantitative polymerase chain reaction. Primary in vitro culture of mouse lung epithelial cells (MLE-12), FIZZ1 recombinant protein and FIZZ1-shRNA interference were used to detect the expression of E-cadherin, type I collagen and fibronectin-1. Results Compared with the control group, the expression of FIZZ1 and FIZZ1 mRNA in the airway epithelium of OVA group was significantly increased, the type Ⅰ collagen and fibronectin 1 were significantly increased, and the expression of E-cadherin was significantly decreased, which was positively and negatively correlated with FIZZ1; In vitro primary culture of MLE-12, FIZZ1 recombinant protein intervention, type I collagen and fibronectin 1 increased, E-cadherin decreased significantly; FIZZ1-shRNA interference, protein expression was normal. Conclusion FIZZ1, as a potential inducer of EMT, induces the decrease of E-cadherin expression and the increase of type I collagen and fibronectin-1, thus inducing early airway remodeling in asthma.