TGF-β1/Smad信号通路相关因子在牦牛肺脏低氧适应中的表达分析  

Expression Analysis of TGF-β1/Smad Signaling Pathway Related Factors During Lung Hypoxia Adaptation in Yak(Bos grunniens)

作　　者：王可金 文德全 崔燕[1] 张倩[1] 何俊峰[1] WANG Ke-Jin;WEN De-Quan;CUI Yan;ZHANG Qian;HE Jun-Feng(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《农业生物技术学报》2023年第1期105-114,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31960687)。

摘　　要：低氧应激下,转化生长因子-β1(transforming growth factors-β1,TGF-β1)/Smad信号通路能被激活并通过促进肺动脉平滑肌细胞的增殖引起低氧性肺动脉高压的发生。为探究TGF-β1/Smad信号通路在牦牛(Bos grunniens)肺脏低氧适应过程中能否被激活,以及其对牦牛肺脏低氧适应性结构形成的可能作用,本研究利用qPCR检测TGF-β1、Smad2和Smad3基因在初生、幼年、成年和老年牦牛肺脏组织中的相对表达量,同时采用Western blot和免疫组织化学方法对初生、幼年、成年和老年牦牛肺脏组织中TGF-β1、Smad2和Smad3的蛋白表达水平,Smad2/Smad3磷酸化蛋白水平(P-Smad2/Smad3)及其分布特征进行研究。qPCR和Western blot结果显示,TGF-β1、Smad2、Smad3和P-Smad2/Smad3的表达趋势基本一致,且在初生、幼年、成年和老年组牦牛肺脏组织中存在表达差异,均在初生和成年组牦牛肺脏组织中表达量高,显著高于幼年和老年组牦牛肺脏组织(P<0.05)。免疫组化结果显示,TGF-β1、Smad2、Smad3和PSmad2/Smad3的分布位置基本一致,主要分布于初生、幼年、成年和老年组牦牛肺脏气管上皮细胞、肺动脉内皮细胞和肺泡Ⅱ型上皮细胞,初生、成年和老年组牦牛肺脏气管壁平滑肌细胞和肺动脉平滑肌细胞也有其阳性表达。本研究发现TGF-β1/Smad信号通路在牦牛肺脏低氧适应过程中被激活,且在初生、幼年、成年和老年组牦牛肺脏组织中的表达活性存在显著差异,初生组表达活性较高,幼年组显著下调,成年组表达活性达最高,老年组又降低,这提示TGF-β1/Smad信号通路活性的动态调控可能参与牦牛肺脏低氧适应性结构的形成和肺血管功能稳定的维持。本研究为进一步研究牦牛肺脏低氧适应机制提供了资料。Under hypoxic stress,transforming growth factors-β1(TGF-β1)/Smad signaling pathway can be activated and caused the occurrence of hypoxic pulmonary hypertension by promoting the proliferation of pulmonary smooth muscle cells.In this study,qPCR was used to detect the relative expression levels of TGF-β1,Smad2 and Smad3 genes in the lung tissues of newborn,juvenile,adult and aged yak(Bos grunniens).The protein expression levels of TGF-β1,Smad2 and Smad3,the phosphorylated protein levels of Smad2/Smad3(P-Smad2/Smad3)and their distribution characteristics in the lung tissues of newborn,juvenile,adult and aged yak were analyzed by Western blot and immunohistochemistry method.The results of q PCR and Western blot showed that the expression trends of TGF-β1,Smad2,Smad3 and P-Smad2/Smad3 were basically the same,and there were differences in expression in the lung tissue of yak in the newborn,juvenile,adult and aged groups,and the expression levels were higher in the lung tissue of the newborn and adult yak groups,which was significantly higher than that in the yak lung tissue of the juvenile and aged groups(P<0.05).The results of immunohistochemistry showed that the distribution of TGF-β1,Smad2,Smad3 and P-Smad2/Smad3 was basically the same,mainly distributed in tracheal epithelial cells,pulmonary artery endothelial cells and alveolar typeⅡepithelial cells in the lungs of newborn,juvenile,adult and aged yaks,the positive expression was also found in lung trachea wall smooth muscle cells and pulmonary artery smooth muscle cells in newborn,adult and aged yaks.This study found that TGF-β1/Smad signal pathway was activated during hypoxic adaptation in yak lungs,and there were significant differences in expression activity among newborn,juvenile,adult and aged yak lungs,the expression activity was higher in the newborn group,significantly down-regulated in the young group,the highest in the adult group,and decreased in the old group.These results showed that the dynamic regulation of TGF-β1/Smad signaling pathway cou

关 键 词：牦牛 肺脏 低氧性肺动脉高压 TGF-Β1/SMAD信号通路 

分 类 号：S852.1[农业科学—基础兽医学]