|
|
|
| Expression Analysis of TGF-β1/Smad Signaling Pathway Related Factors During Lung Hypoxia Adaptation in Yak (Bos grunniens) |
| WANG Ke-Jin, WEN De-Quan, CUI Yan, ZHANG Qian, HE Jun-Feng* |
| College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China |
|
|
|
|
Abstract 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 qPCR 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 type Ⅱ epithelial 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 could be involved in the formation of hypoxia adaptive structure and the maintenance of pulmonary vascular function in yaks. The results provide data for further study on the mechanism of lung hypoxia adaptation in yaks.
|
|
Received: 15 February 2022
|
|
|
|
Corresponding Authors:
*hejf@gsau.edu.cn
|
|
|
|
[1] 何俊峰, 余四九, 崔燕. 2009. 不同年龄高原牦牛肺脏的组织结构特征[J]. 畜牧兽医学报, 40(5): 748-755.
(He J F, Yu S J, Cui Y.2009. The tissue structure characteristics of the lungs of plateau yak at different ages[J]. Journal of Animal Husbandry and Veterinary Medicine, 40(05): 748-755.)
[2] 靳新花, 崔燕, 何俊峰, 等. 2009. 成年牦牛肺动脉的显微结构观察[J]. 中国兽医科学, 39(03): 261-265.
(Jin X H, Cui Y, He J F, et al.2009. Microstructural observation of adult yak pulmonary artery[J]. Veterinary Science in China, 39(03): 261-265.)
[3] 李怡, 李学智, 廖冬梅, 等. 2022. 基于MCP-1/MMP-2/TGF-β1局部促炎信号环探讨温和灸对老年大鼠血管衰老的干预作用[J]. 针刺研究, 47(04): 290-297.
(Li Y, Li X Z, Liao D M, et al.2022. To investigate the intervention effect of mild moxibustion on vascular aging in aged rats based on the local pro-inflammatory signal loop of MCP-1/MMP-2/TGF-β1[J]. Acupuncture Research, 47(04): 290-297.)
[4] 文德全, 崔燕, 余四九, 等. 2022. HIF-1α和FGF-2在不同年龄牦牛睾丸中的表达与定位[J]. 中国兽医科学, 52(06): 789-796.
(Wen D Q, Cui Y, Yu S J, et al.2022. Expression and localization of HIF-1α and FGF-2 in yak testis of different ages[J]. Veterinary Science in China, 52(06): 789-796.)
[5] 杨超, 丁学智, 钱娇玲, 等. 2017. 牦牛适应青藏高原环境的组织解剖学研究进展[J]. 中国畜牧杂志, 53(03): 18-24.
(Yang C, Ding X Z, Qian J L, et al.2017. Research progress on the anatomy of yak adapted to the environment of the Qinghai-Tibet Plateau[J]. Chinese Journal of Animal Science, 53(03): 18-24.)
[6] 杨琨, 余四九, 何俊峰, 等. 2012. 牦牛胎儿肺脏发育的形态学研究[J]. 兽类学报, 32(04): 346-355.
(Yang K, Yu S J, He J F, et al.2012. Morphological study of yak fetal lung development[J]. Acta Theriologica Sinica, 32(04): 346-355.)
[7] 张兰, 姚一凡, 覃安琪, 等. 2022. 基于TMT技术对低氧与常氧条件下牦牛PASMCs的蛋白组学分析[J]. 畜牧兽医学报, 53(07): 2182-2193.
(Zhang L, Yao Y F, Tan A Q, et al.2022. Proteomic analysis of yak PASMCs under hypoxic and normoxic conditions based on TMT technology[J]. Journal of Animal Husbandry and Veterinary Medicine, 53(07): 2182-2193.)
[8] 张思源, 柴志欣, 钟金城. 2016. 牦牛高原低氧适应研究进展[J]. 江苏农业科学, 44(03): 13-17.
(Zhang S Y, Chai Z X, Zhong J C, et al.2016. Research progress of yak plateau hypoxia adaptation[J]. Jiangsu Agricultural Sciences, 44(03): 13-17.)
[9] Andre P, Joshi S R, Briscoe S D, et al.2021. Therapeutic approaches for treating pulmonary arterial hypertension by correcting imbalanced TGF-β superfamily signaling[J]. Frontiers in Medicine, 8: 814222.
[10] Alejandre-Alcázar M A, Michiels-Corsten M, Vicencio A G, et al.2008. TGF-beta signaling is dynamically regulated during the alveolarization of rodent and human lungs[J]. Developmental Dynamics, 237(1): 259-269.
[11] Ambalavanan N, Nicola T, Hagood J, et al.2008. Transforming growth factor-beta signaling mediates hypoxia-induced pulmonary arterial remodeling and inhibition of alveolar development in newborn mouse lung[J]. American Journal of Physiology Lung Cellular and Molecular Physiology, 295(1): L86-95.
[12] Chai S D, Liu T, Dong M F, et al.2016. Inactivated pseudomonas aeruginosa inhibits hypoxia-induced pulmonary hypertension by preventing TGF-β1/Smad signaling[J]. Brazilian Journal of Medical and Biological Research, 49(10): e5526.
[13] Chapman H A.2011. Epithelial-mesenchymal interactions in pulmonary fibrosis[J]. Annual Review of Physiology, 73: 413-435.
[14] Chen L, Ge Q, Black J L, et al.2013. Differential regulation of extracellular matrix and soluble fibulin-1 levels by TGF-β1 in airway smooth muscle cells[J]. PLOS ONE, 8(6): e65544.
[15] Chen Y F, Feng J A, Li P, et al.2006. Dominant negative mutation of the TGF-beta receptor blocks hypoxia-induced pulmonary vascular remodeling[J]. Journal of Applied Physiology, 100(2): 564-571.
[16] Ding F, You T, Hou X D, et al.2019. MiR-21 regulates pulmonary hypertension in rats via TGF-β1/Smad2 signaling pathway[J]. European Review for Medical and Pharmacological Sciences, 23(9): 3984-3992.
[17] Doerr M, Morrison J, Bergeron L, et al.2016. Differential effect of hypoxia on early endothelial-mesenchymal transition response to transforming growth beta isoforms 1 and 2[J]. Microvascular Research, 108: 48-63.
[18] Durmowicz A G, Hofmeister S, Kadyraliev T K, et al.1993. Functional and structural adaptation of the yak pulmonary circulation to residence at high altitude[J]. Journal of Applied Physiology, 74(5): 2276-2285.
[19] Gao W, Shao R, Zhang X, et al.2017. Up-regulation of caveolin-1 by DJ-1 attenuates rat pulmonary arterial hypertension by inhibiting TGFβ/Smad signaling pathway[J]. Experimental Cell Research, 361(1): 192-198.
[20] Ge R L, Kubo K, Kobayashi T, et al.1998. Blunted hypoxic pulmonary vasoconstrictive response in the rodent Ochotona curzoniae (pika) at high altitude[J]. The American Journal of Physiology, 274(5): H1792-1799.
[21] Hu H H, Chen D Q, Wang Y N, et al.2018. New insights into TGF-β/Smad signaling in tissue fibrosis[J]. Chemico-Biological Interactions, 292: 76-83.
[22] Huang S, Chen P, Shui X, et al.2014. Baicalin attenuates transforming growth factor-β1-induced human pulmonary artery smooth muscle cell proliferation and phenotypic switch by inhibiting hypoxia inducible factor-1α and aryl hydrocarbon receptor expression[J]. The Journal of Pharmacy and Pharmacology, 66(10): 1469-1477.
[23] Ishizaki T, Koizumi T, Ruan Z, et al.2005. Nitric oxide inhibitor altitude-dependently elevates pulmonary arterial pressure in high-altitude adapted yaks[J]. Respiratory Physiology & Neurobiology, 146(2-3): 225-230.
[24] Ishizaki T, Mizuno S, Sakai A, et al.2015. Blunted activation of Rho-kinase in yak pulmonary circulation[J]. BioMed Research International, 2015: 720250.
[25] Lan D, Xiong X, Ji W, et al.2018. Transcriptome profile and unique genetic evolution of positively selected genes in yak lungs[J]. Genetica, 146(2): 151-160.
[26] Xu M Y, Pang Q Q, Xu S Q, et al.2018. Hypoxia-inducible factor-1α activates transforming growth factor-β1/Smad signaling and increases collagen deposition in dermal fibroblasts[J]. Oncotarget, 9(3): 3188-3197.
[27] Pak O, Aldashev A, Welsh D, et al.2007. The effects of hypoxia on the cells of the pulmonary vasculature[J]. The European Respiratory Journal, 30(2): 364-372.
[28] Qi X, Zhang Q, He Y, et al.2019. The transcriptomic landscape of yaks reveals molecular pathways for high altitude adaptation[J]. Genome Biology and Evolution, 11(1): 72-85.
[29] Rabinovitch M, Gamble W J, Miettinen O S, et al.1981. Age and sex influence on pulmonary hypertension of chronic hypoxia and on recovery[J]. The American Journal of Physiology, 240(1): H62-72.
[30] Richter A, Yeager M E, Zaiman A, et al.2004. Impaired transforming growth factor-beta signaling in idiopathic pulmonary arterial hypertension[J]. American Journal of Respiratory and Critical Care Medicine, 170(12): 1340-1348.
[31] Stenmark K R, Fagan K A, Frid M G.2006. Hypoxia-induced pulmonary vascular remodeling: Cellular and molecular mechanisms[J]. Circulation Research, 99(7): 675-691.
[32] Van Loon K, Yemelyanenko-Lyalenko J, Margadant C, et al.2020. Role of fibrillin-2 in the control of TGF-β activation in tumor angiogenesis and connective tissue disorders[J]. Biochimica et Biophysica Acta[J]. Reviews on Cancer, 1873(2): 188354.
[33] Wang X B, Wang W, Zhu X C, et al.2015. The potential of asiaticoside for TGF-β1/Smad signaling inhibition in prevention and progression of hypoxia-induced pulmonary hypertension[J]. Life Sciences, 137: 56-64.
[34] Weber R E, Lalthantluanga R, Braunitzer G.1988. Functional characterization of fetal and adult yak hemoglobins: An oxygen binding cascade and its molecular basis[J]. Archives of Biochemistry & Biophysics, 263(1): 199-203.
[35] Yue Y, Li Y Q, Fu S, et al.2020. Sthole inhibits cell proliferation by regulating the TGF-β1/Smad/p38 signaling pathways in pulmonary arterial smooth muscle cells[J]. Biomedicine & Pharmacotherapy, 121: 109640.
[36] Zhang H, He H, Cui Y, et al.2021. Regulatory effects of HIF-1α and HO-1 in hypoxia-induced proliferation of pulmonary arterial smooth muscle cells in yak[J]. Cellular Signalling, 87: 110140.
[37] Zhang N, Dong M, Luo Y, et al.2018. Danshensu prevents hypoxic pulmonary hypertension in rats by inhibiting the proliferation of pulmonary artery smooth muscle cells via TGF-β-smad3-associated pathway[J]. European Journal of Pharmacology, 820: 1-7.
[38] Zhao R, Li N, Xu J, et al.2018. Quantitative single-molecule study of TGF-β/Smad signaling[J]. Acta Biochimica et Biophysica Sinica, 50(1): 51-59.
[39] Zhou J, Yu S, He J, et al.2013. Segmentation features and structural organization of the intrapulmonary artery of the yak[J]. Anatomical Record, 296(11): 1775-1788. |
|
|
|
