【摘要】 目的:观察降钙素基因相关肽(CGRP)对大鼠肺泡上皮细胞间质转分化(EMT)的作用并探讨其机制。方法:实验设Control组、TGF-β1(5 ng/ml)、CGRP(1、10、100 nmol/L)组、CGRP8-37(1μmol/L)+CGRP(100nmol/L)组。每组设9个复孔。细胞分别用CGRP或加CGRP8-37预处理1 h,再用转化生长因子β1(TGF-β1)处理48 h。MTT法检测大鼠肺泡上皮Ⅱ型细胞(RLE-6TN细胞)活性。分别采用Real-time PCR和Western blot检测细胞E-cadherin、α-SMA、e IF3a、Notch1和collagenⅢmRNA及蛋白表达。结果:与TGF-β1(5 ng/ml)相比,不同剂量CGRP(1、10、100 nmol/L)均可明显提高RLE-6TN细胞活性,显著抑制e IF3a、Notch1、α-SMA及collagenⅢ胶原的表达,上调E-cadherin的表达,而CGRP的这些作用可以被CGRP阻断剂CGRP8-37所取消。结论:CGRP对EMT具有一定的抑制作用,其机制可能与其抑制Notch1、e IF3a表达有关。更多还原

【Abstract】 Objective: To observe the effects of calcitonin gene-related peptide( CGRP) on epithelial-mesenchymal transition( EMT) of alveolar epithelial cells and its mechanism. Methods: The RLE-6 TN cell were divided into 6 groups as follows: control group,cells were incubated with double distilled water( TGF-β1 solvent) for 48 h; transforming growth factor-β1( TGF-β1) group,cells were incubated with TGF-β1( 5 ng/ml) for 48 h; + CGRP( 1,10,100 nmol/L) group,cells were pre-treated with CGRP( 1,10,100 nmol/L) for 1 h,and then subjected to TGF-β1 for 48 h; CGRP 100 nmol/L + CGRP8-37 1 μmol/L group,cells were pretreated with CGRP 100 nmol/L + CGRP8-37 1 μmol/L for 1 h,and then subjected to TGF-β1 for 48 h. RLE-6 TN cell vitality was determined by MTT. The mRNA or protein expression levels of Notch1,e IF3 a,α-SMA,E-cadherin and collagen Ⅲ were detected by real-time PCR or Western blot. Results: Compared with the TGF-β1 group,cell vitality and the expression of E-cadherin were significantly increased,and the expression of Notch 1,e IF3 a,α-SMA and collagen Ⅲ were markedly decreased in CGRP( 1,10,100 nmol/L) group. The effects of CGRP above mentioned could be abolished by CGRP 8-37. Conclusion: These results suggest that CGRP inhibits TGF-β1-induced EMT process,and down-regulation of the expressions of Notch1 and e IF3 a may be involved in its mechanisms.更多还原