降钙素基因相关肽对大鼠肺泡上皮Ⅱ型细胞间质转分化的作用及机制  

Effects of calcitonin gene-related peptide on epithelial-mesenchymal transition of alveolar epithelial cells in rat and its mechanism

作　　者：李先伟[1] 蒋莉莉[1] 左东泽 郝伟[1] 吴海龙[2] LI Xian-wei;JIANG Li-li;ZUO Dong-ze;HAO Wei;WU Hai-long(Department of Pharmacology,Wannan Medical College,Anhui Provincial Engineering Technology Research Center for Polysaccharide Drugs,Wuhu 241002;College of Life Sciences,Anhui Normal University,Wuhu 241000,China)

机构地区：[1]皖南医学院药理学教研室安徽省多糖药物工程技术研究中心,芜湖241002 [2]安徽师范大学生命科学学院,芜湖241000

出　　处：《中国应用生理学杂志》2018年第3期249-253,共5页Chinese Journal of Applied Physiology

基　　金：安徽省高校优秀青年人才支持计划重点项目(gxyq ZD2016170)

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。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.

关 键 词：降钙素基因相关肽 上皮细胞间质转分化 eIF3a NOTCH1 大鼠 

分 类 号：R966[医药卫生—药理学]