多配体蛋白多糖-1在慢性阻塞性肺疾病大鼠肺上皮间质转化中的作用及机制*

doi:10.12047/j.cjap.6001.2020.128

中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (6): 611-615.doi: 10.12047/j.cjap.6001.2020.128

多配体蛋白多糖-1在慢性阻塞性肺疾病大鼠肺上皮间质转化中的作用及机制^*

金艳¹, 刘文明²

1.南京医科大学附属常州第二人民医院急诊内科, 江苏常州 213000;
2.南京医科大学附属常州第二人民医院重症监护室, 江苏常州 213000

收稿日期:2020-02-26 修回日期:2020-11-25 出版日期:2020-11-28 发布日期:2021-03-15
通讯作者: ^△Tel: 13915056315; E-mail: lwmicu@163.com
基金资助:
^*河南省新乡医学院第一附属医院青年培育基金项目(QN-2017-B027)

The role of syndecan-1 in the transformation of pulmonary epithelial stroma in rats with chronic obstructive pulmonary disease and its mechanism

JIN Yan¹, LIU Wen-ming²

1. Department of Emergency Medicine, Changzhou 213000, China;
2. ICU, Second People's Hospital of Changzhou Affiliated to Nanjing Medical University, Changzhou 213000, China

Received:2020-02-26 Revised:2020-11-25 Online:2020-11-28 Published:2021-03-15

摘要/Abstract

摘要： 目的:探讨多配体蛋白多糖-1(syndecan-1)在慢性阻塞性肺疾病(COPD)大鼠肺上皮间质转化(EMT)中的作用及机制。方法:选择清洁级SD雄性大鼠30只,随机分为假手术组(暴露气管后注射生理盐水,n=10),COPD组(注射1 mg/ml脂多糖后进行烟熏,造模完成后转染100 μl空载病毒,n=10),syndecan-1过表达组(注射1 mg/ml脂多糖后进行烟熏。造模完成后转染100 μl携带大鼠syndecan-1基因的重组腺病毒载体Ad-CMV-GFP-SDC1,n=10),每天1次,连续处理2周。处理结束后检测各组大鼠肺功能并取肺组织;采用HE染色法观察肺组织损伤情况;采用免疫组化检测各组大鼠肺组织syndecan-1、波形蛋白(vimentin)、E-钙黏蛋白(E-cadherin)的蛋白表达;采用Western blot检测转化生长因子-β1(TGF-β1)、Smad2/3和p-Smad2/3蛋白表达水平;采用qRT-PCR法检测各组大鼠肺组织中vimentin、E-cadherin、TGF-β1和Smad2/3 mRNA表达水平。结果:与假手术组相比,COPD组大鼠气道黏膜脱落,管腔狭窄,气道壁较多炎性细胞浸润,肺气肿严重,每分钟呼气量(V_E)、最大呼气流量(PEF)和0.3s用力呼气容积(FEV_0.3)和肺组织E-cadherin mRNA表达水平均显著降低(P＜0.05),而vimentin、TGF-β1、Smad2/3 mRNA表达水平及TGF-β1、Smad2/3和p-Smad2/3的蛋白表达水平均显著升高(P＜0.05)。与COPD组相比,syndecan-1过表达组气道黏膜脱落及气道壁炎性细胞浸润数减少,管腔狭窄、肺气肿情况均有所改善,V_E、PEF、FEV_0.3和肺组织E-cadherin mRNA表达水平显著升高(P＜0.05),而vimentin、TGF-β1、Smad2/3 mRNA表达水平及TGF-β1、Smad2/3和p-Smad2/3的蛋白表达水平均显著降低(P＜0.05)。结论:COPD大鼠体内TGF-β/Smad信号通路活化且存在肺EMT;过表达syndecan-1可抑制TGF-β/Smad信号通路,减轻EMT,改善COPD大鼠肺组织损伤。

关键词: 多配体蛋白多糖-1, 慢性阻塞性肺疾病, 大鼠, 上皮间质转化

Abstract: Objective: To explore the role and mechanism of polysaccharide-1 (syndecan-1) in the transformation of lung epithelial stroma (EMT) in rats with chronic obstructive pulmonary disease (COPD). Methods: Thirty male SD rats of clean grade were randomly divided into sham operation group (normal saline injection after tracheal exposure, n=10), COPD group (fumigation after injection of 1 mg/ml lipopolysaccharide, transfection of 100 μl empty virus, n=10) and syndecan-1 overexpression group (fumigation after injection of 1 mg/ml lipopolysaccharide, and transfection of 100 μl carrying rat syndecan-1 gene Ad-CMV-GFP-SDC1, n=10), once a day for two weeks. After the treatment, the lung function was detected and lung tissues were collected. HE staining was used to observe lung injury. The expression levels of syndecan-1, vimentin and E-cadherin in lung tissue of rats in each group were detected by immunohistochemistry. Western blot was used to detect the expressions of TGF-β1, Smad2/3 and p-Smad2/3. The mRNA levels of vimentin, E-cadherin, TGF-β1 and Smad2/3 were detected by qRT-PCR. Results: Compared with the sham operation group, the airway mucosa of COPD group was exfoliated, the lumen was narrow, the airway wall was more inflammatory cell infiltration, emphysema was serious, expiratory volume (V_E), peak expiratory flow (PEF), Forced expiratory volume in 0.3 s (FEV_0.3) and the expression of E-cadherin mRNA were significantly decreased (P＜0.05). While the expression of vimentin, TGF-β1 and Smad2/3 mRNA were increased, and the expression levels of TGF-β1, Smad2/3 and p-Smad2/3 protein were significantly increased (P＜0.05). Compared with COPD group, the number of airway mucosa exfoliation and inflammatory cell infiltration in airway wall were decreased, lumen stenosis and emphysema were improved, the levels of V_E, PEF, FEV_0.3 and the expression of E-cadherin mRNA were significantly increased (P＜0.05). And the expressions of vimentin, TGF-β1 and Smad2/3 mRNA were decreased, and expression levels of TGF-β1, Smad2/3 and p-Smad2/3 protein were significantly decreased (P＜0.05). Conclusion: In COPD rats, TGF-β/Smad signal pathway activation induced the production of EMT; overexpression of syndecan-1 could inhibit the EMT mediated by TGF-β/Smad signal pathway, and improve the lung tissue injury of COPD rats.

Key words: syndecan-1, COPD, rat, EMT

中图分类号:

金艳, 刘文明. 多配体蛋白多糖-1在慢性阻塞性肺疾病大鼠肺上皮间质转化中的作用及机制^*[J]. 中国应用生理学杂志, 2020, 36(6): 611-615.

JIN Yan, LIU Wen-ming. The role of syndecan-1 in the transformation of pulmonary epithelial stroma in rats with chronic obstructive pulmonary disease and its mechanism[J]. CJAP, 2020, 36(6): 611-615.

参考文献

[1] 王晓娟, 方向阳. 慢性阻塞性肺疾病全球倡议2019:慢性阻塞性肺疾病诊断、治疗与预防全球策略解读[J]. 中国全科医学, 2019, 22(18): 2141-2149.
[2] Bartis D, Mise N, Mahida RY, et al. Epithelial-mesenchymal transition in lung development and disease: does it exist and is it important[J] ? Thorax, 2014, 69(8): 760-765.
[3] Charchanti A, Papoudou Bai A, Samantas E, et al. Association of low Syndecan-1 expression with adverse histopathological parameters in gastric carcinomas[J]. J BUON, 2019, 24(3): 1106-1112.
[4] Li D, Wu Y, Guo S, et al. Circulating syndecan-1 as a novel biomarker relates to lung function, systemic inflammation, and exacerbation in COPD[J]. Int J Chron Obstruct Pulmon Dis, 2019, 14: 1933-1941.
[5] 黄文锋, 陈斯宁, 吴嘉冬, 等. 慢性阻塞性肺疾病大鼠模型的构建[J]. 现代医药卫生, 2019, 35(6): 801-803.
[6] 陈启华, 王改丽, 刘文龙. 辛伐他汀对慢性阻塞性肺疾病模型大鼠气道炎症和气道黏液高分泌的防治作用及其机制[J]. 中华医学杂志, 2015, 95(22): 1726-1730.
[7] 于娜, 陈建明, 李晓才, 等. SIRT1对肺癌A549细胞增殖和迁移的影响[J]. 广东医学, 2017, 38(1): 65-71.
[8] Sohal SS, Walters EH. Role of epithelial mesenchymal transition(EMT)in chronic obstructive pulmonary disease (COPD)[J]. Respir Res, 2013, 14(1): 120-123.
[9] Zhou J, Tao D, Xu Q, et al. Expression of E-cadherin and vimentin in oral squamous cell carcinoma[J]. Int J Clin Exp Pathol, 2015, 8(3): 3150-3154.
[10] 周立莉, 葛新国, 蔡茂怀, 等. 非小细胞肺癌中ETS-1与E-cadherin、N-cadherin、Vimentin表达相关性的研究[J]. 癌症进展, 2015, 13(5): 504-511.
[11] Dong QZ, Zhu X, Dai C, et al. Osteopontin promotes epithelial-mesenchymal transition of hepatocellular carcinoma through regulating vimentin[J]. Oncotarget, 2016, 7(11): 12997-13012.
[12] 杨欣, 陈睿, 吴晨, 等. 过表达磷脂酰肌醇蛋白聚糖-5对肺腺癌上皮-间充质转化的影响及临床意义[J]. 中华实验外科杂志, 2018, 35(10): 1800-1802.
[13] 许家艳, 孟亚奇, 贾嫚, 等. 人支气管上皮细胞表达的斯钙素-1在上皮间充质转化中的作用研究[J]. 南京医科大学学报(自然科学版), 2018, 38(9): 1198-1203.
[14] 张云珍, 方海燕, 余红, 等. 白藜芦醇抑制TGF-β1/Smad3信号通路对人肺上皮细胞A549 EMT调控的影响及机制[J]. 广东医学, 2019, 40(15): 2130-2134.
[15] 孔晓武, 丁青, 张琦. AMPK抑制TGF-β信号通路减轻EMT诱导膀胱癌转移机制研究[J]. 中国现代医生, 2019, 57(12): 28-31.
[16] 代炜, 姚远, 李彦姝, 等. TGF-β1上调MTA1促进人舌癌Tca8113细胞侵袭转移[J]. 现代肿瘤医学, 2013, 21(5): 950-952.
[17] Fujii T, Shimada K, Tatsumi Y, et al. Syndecan-1 up-regulates microRNA-331-3p and mediates epithelial-to-mesenchymal transition in prostate cancer[J]. Mol Carcinog, 2016, 55(9): 1378-1386.
[18] 许丽娜, 赵静, 应仙华. 子宫内膜异位症患者血清syndecan-1及MMP-9水平变化及其临床意义[J]. 中华内分泌外科杂志, 2018, 12(4): 326-329.

多配体蛋白多糖-1在慢性阻塞性肺疾病大鼠肺上皮间质转化中的作用及机制^*

The role of syndecan-1 in the transformation of pulmonary epithelial stroma in rats with chronic obstructive pulmonary disease and its mechanism

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	徐云, 陈雪辉, 白立炜, 尹清风, 冯春瑜, 张清贵. 达格列净对2型糖尿病大鼠肾脏葡萄糖转运蛋白2及葡萄糖转运蛋白4基因表达的影响^*[J]. 中国应用生理学杂志, 2020, 36(6): 565-570.
[2]	王丹, 高峰, 陈慧, 项伟玲, 骆志显, 金丽琴. 人参归脾丸对脾不统血证大鼠肝损伤的保护作用及其机制^*[J]. 中国应用生理学杂志, 2020, 36(6): 571-575.
[3]	韦凌霞, 丁茂鹏, 王志旺, 刘雪枫, 庞亚蓉, 郭玫. 鳖甲育肝颗粒对乙醇性肝纤维化大鼠的影响^*[J]. 中国应用生理学杂志, 2020, 36(6): 582-586.
[4]	张冉, 马梦尧, 苏欣宇, 孟想, 姜鲲鹏, 李曙, 洪云. 大鼠脑缺血/再灌注过程中血流量和脑组织含水量的变化趋势^*[J]. 中国应用生理学杂志, 2020, 36(6): 586-589.
[5]	苏海容, 程文姚, 袁秋虹, 欧阳菁, 邓伟民. 补肾壮骨颗粒对去卵巢大鼠血清GH和IGF-1及其骨组织中受体表达的影响^*[J]. 中国应用生理学杂志, 2020, 36(6): 605-610.
[6]	冉卉, 尹浩, 刘窗溪, 韩国强, 高方友, 谌鸿斌, 付航, 徐晓钟, 黎涛, 马骏. 甲状腺素对动脉瘤性蛛网膜下腔出血大鼠脑缺氧诱导因子-1α表达的影响及其机制^*[J]. 中国应用生理学杂志, 2020, 36(6): 648-652.
[7]	孙科, 罗招亮, 胡琛, 龚廷亮, 唐国华, 吴绍萍. 小檗碱对大鼠脑缺血/再灌注损伤的保护作用及免疫机制^*[J]. 中国应用生理学杂志, 2020, 36(6): 656-661.
[8]	潘文强, 王书凡, 丁伯平, 黄帧桧. 格列齐特对糖尿病大鼠心肌的保护作用及其机制^*[J]. 中国应用生理学杂志, 2020, 36(5): 402-407.
[9]	高丽, 张文慧, 王羽维, 朱晓佳, 肖钰洁, 高倩雯, 李伟, 陈龙. 苍术素对大鼠心脏正性肌力的作用及其机制^*[J]. 中国应用生理学杂志, 2020, 36(5): 408-413.
[10]	李杰茹, 任静, 杨胜昌, 韩聚强, 郭亚净, 吉恩生. 消痰化瘀利窍中药组方对改善慢性间歇性低氧大鼠心肌纤维化的作用及其机制^*[J]. 中国应用生理学杂志, 2020, 36(5): 414-418.
[11]	贾祥磊, 刘玉珍, 李超红, 范娅楠. 多巴胺对急性间歇性低氧诱导的大鼠颈动脉体低氧敏感性的影响^*[J]. 中国应用生理学杂志, 2020, 36(5): 428-431.
[12]	谢亚磊, 梅颂, 熊艳蕾, 刘诗颖, 徐成丽. 模拟海拔5 500 m低压低氧环境对大鼠HPT轴和肠道菌群的影响^*[J]. 中国应用生理学杂志, 2020, 36(5): 432-437.
[13]	郭锐, 秦卫帅, 张双勇, 张淑玲. 甘麦大枣汤对抑郁模型大鼠海马神经元突触结构及结构蛋白表达的影响^*[J]. 中国应用生理学杂志, 2020, 36(5): 444-448.
[14]	孙林, 柏友萍, 林伟其. 脂肪因子Vaspin在运动与甲鱼油对老年肥胖大鼠胰岛素抵抗的作用^*[J]. 中国应用生理学杂志, 2020, 36(5): 461-464.
[15]	刘玉倩, 张静, 高丽娜, 王海涛. 有氧运动对高糖高脂膳食大鼠腓肠肌Nrf2-SOD的影响^*[J]. 中国应用生理学杂志, 2020, 36(5): 481-485.