膜联蛋白A1促进肺成纤维细胞炎症反应及胶原合成的研究

蔡欣; 李炎玉; 苏国媚; 吕秋荣; 赖天文

doi:10.16766/j.cnki.issn.1674-4152.002172

膜联蛋白A1促进肺成纤维细胞炎症反应及胶原合成的研究

doi: 10.16766/j.cnki.issn.1674-4152.002172

广东医科大学附属医院呼吸与危重症医学科，广东湛江 524001

基金项目:

广东省医学科研基金项目 A2018162

详细信息

通讯作者:
赖天文，E-mail：laitianwen2011@163.com

中图分类号: R563 R-332
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出版历程
- 收稿日期: 2020-12-25
- 网络出版日期: 2022-02-15

Effects of Annexin A1 on inflammatory response and collagen deposition of lung fibroblast

Department of Respiratory and Critical Care Medicine, the Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, China

摘要

摘要: 目的探讨膜联蛋白A1(Annexin A1)对肺成纤维细胞炎症反应、增殖及胶原沉积的作用。方法分离小鼠原代肺成纤维细胞，利用不同浓度的Annexin A1干预肺成纤维细胞24 h或48 h后，检测炎症因子、细胞增殖及胶原沉积表达。将C57/BL6小鼠分为对照组、Annexin A1干预组，每组6~8只。Annexin A1干预组每只小鼠气管滴注1 μg/50 μL Annexin A1重组因子，对照组每只小鼠气道滴注50 μL生理盐水，连续干预7 d后取检。检测肺组织炎症因子、Ⅲ型胶原(Collagen Ⅲ)及气道平滑肌肌动蛋白(α-SMA)的表达。结果与对照组比较，Annexin A1能促进肺成纤维细胞分泌IL-6、IL-8等炎症因子(均P < 0.05)，促进成纤维细胞的增殖，Collagen Ⅲ合成和α-SMA表达增加(均P < 0.05)。与对照组小鼠比较，Annexin A1干预组小鼠的肺泡灌洗液细胞总数[(2.91±0.26)×10⁴/mL vs.(7.03±0.48)×10⁴/mL，t=7.432，P=0.008]及中性粒细胞数目明显升高[(0.12±0.04)×10⁴/mL vs. (1.01±0.05)×10⁴/mL，t=13.810，P < 0.001]，炎症因子、胶原沉积及平滑肌表达也显著增加(均P < 0.05)。结论 Annexin A1具有诱导肺成纤维细胞的炎症反应、增殖及胶原沉积作用，提示可能在气道重构发病中起重要作用。
- 膜联蛋白A1 /
- 肺成纤维细胞 /
- 肺纤维化
Abstract: Objective To investigate the effects of Annexin A1 on inflammatory response, proliferation and collagen deposition of pulmonary fibroblasts. Methods The primary lung fibroblasts of C57/BL6 mice were isolated. After 24 or 48 hours of intervention with different concentrations of Annexin A1, the expression of inflammatory factors, cell proliferation and collagen deposition were detected. C57/BL6 mice were randomly divided into control group and Annexin A1 intervention group according to the random number table method, with 6-8 mice in each group. Each mouse in the Annexin A1 intervention group was instilled with mouse recombinant Annexin A1 (1 μg/50 μL) into the trachea, and each mouse in the control group was instilled with 50 μL saline. After continuous intervention for 7 days, mice were sacrificed for measurement. Inflammatory factors, collagen Ⅲ and airway smooth muscle actin (α-SMA) were detected. Results Inflammatory cytokines (IL-6, IL-8), proliferation, collagen Ⅲ, and α-SMA were significantly increased in the Annexin A1 intervention group compared with the control group (all P < 0.05). Compared with control mice, the total number of cells in the alveolar lavage fluid of the Annexin A1 intervention group [(2.91±0.26)×10⁴/mL vs. (7.03±0.48)×10⁴ /mL, t=7.432, P=0.008] and the number of neutrophils [(0.12±0.04)×10⁴/mL vs. (1.01±0.05)×10⁴/mL, t=13.810, P < 0.001] were significantly increased, and the expression of inflammatory cytokines, collagen deposition, and smooth muscle were also significantly increased (all P < 0.05). Conclusion Annexin A1 induces inflammation, proliferation and collagen deposition in the lung fibroblasts, suggesting that Annexin A1 maybe involved in the pathogenesis of airway remodeling.
- Annexin A1 /
- Pulmonary fibroblasts /
- Pulmonary fibrosis

HTML全文

图 1 Annexin A1诱导小鼠原代肺成纤维细胞炎症因子表达

注：用RT-PCR检测IL-6、IL-8、IL-17A及IL-1β表达。

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图 2 Annexin A1促进小鼠肺组织炎症反应

注：用1 μg/50 μL Annexin A1重组因子滴注小鼠气管，连续7 d。观察小鼠肺泡灌洗液(BALF)细胞总数(A)和中性粒细胞比例(B)。HE观察小鼠肺组织炎症浸润(红色箭头)(C，×200倍)及进行HE染色气道炎症评分(D)。用RT-PCR检测小鼠肺组织炎症因子IL-6及IL-8表达(E、F)。每次实验每组小鼠6~8只，代表 3次独立实验。

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表 1 小鼠定量逆转录聚合酶链反应引物序列

待检测物	引物序列
Annexin A1	Forward Primer AAGCAGGCCCGTTTTCTTGAA
	Reverse Primer GCAACATCCGAGGATACATTGA
IL-6	Forward Primer CTGCAAGAGACTTCCATCCAG
	Reverse Primer AGTGGTATAGACAGGTCTGTTGG
IL-17A	Forward Primer TCAGCGTGTCCAAACACTGAG
	Reverse Primer CGCCAAGGGAGTTAAAGACTT
IL-1β	Forward Primer GAAATGCCACCTTTTGACAGTG
	Reverse Primer TGGATGCTCTCATCAGGACAG
Collagen Ⅲ	Forward Primer CTGTAACATGGAAACTGGGGAAA
	Reverse Primer CCATAGCTGAACTGAAAACCACC
α-SMA	Forward Primer CCCAGACATCAGGGAGTAATGG
	Reverse Primer TCTATCGGATACTTCAGCGTCA

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