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COPD患者肺泡灌洗液来源外泌体通过miR-223-3p/FOXO3通路抑制成骨细胞成骨分化的作用
COPD patients' bronchoalveolar lavage fluid-derived exosomes inhibit osteoblast differentiation
via miR-223-3p/FOXO3 pathway- 陆军军医大学学报 2024年46卷第15期 页码：1780-1788
- 作者机构：
1.400038 重庆，陆军军医大学(第三军医大学)第一附属医院：重症医学科
2.400038 重庆，陆军军医大学(第三军医大学)第一附属医院运动医学中心
- 作者简介：
[陈墨龙，E-mail: chenmlorthop@tmmu.edu.cnCHEN Molong, E-mail: chenmlorthop@tmmu.edu.cn
- 基金信息：
DOI：10.16016/j.2097-0927.202402031
中图分类号： R322.71;R394.3;R563.9收稿：2024-02-07，
修回：2024-06-08，
纸质出版：2024-08-15
- 稿件说明：
移动端阅览
陈红娅, 唐渝镇, 曹英, 等. COPD患者肺泡灌洗液来源外泌体通过miR-223-3p/FOXO3通路抑制成骨细胞成骨分化的作用[J]. 陆军军医大学学报, 2024,46(15):1780-1788.
Hongya CHEN, Yuzhen TANG, Ying CAO, et al. COPD patients' bronchoalveolar lavage fluid-derived exosomes inhibit osteoblast differentiation
via miR-223-3p/FOXO3 pathway[J]. Journal of Army Medical University, 2024, 46(15): 1780-1788.陈红娅, 唐渝镇, 曹英, 等. COPD患者肺泡灌洗液来源外泌体通过miR-223-3p/FOXO3通路抑制成骨细胞成骨分化的作用[J]. 陆军军医大学学报, 2024,46(15):1780-1788. DOI： 10.16016/j.2097-0927.202402031.
Hongya CHEN, Yuzhen TANG, Ying CAO, et al. COPD patients' bronchoalveolar lavage fluid-derived exosomes inhibit osteoblast differentiation
via miR-223-3p/FOXO3 pathway[J]. Journal of Army Medical University, 2024, 46(15): 1780-1788. DOI： 10.16016/j.2097-0927.202402031.
摘要
目的
2
探索慢性阻塞性肺病患者肺泡灌洗液来源的外泌体(COPD-bronchoalveolar lavage fluid-exosome，COPD-Exo)调控成骨细胞分化的作用及机制。
方法
2
选取2023年6月于陆军军医大学第一附属医院就诊的6例COPD患者和6例非COPD患者(对照)，在临床肺泡灌洗过程中收集肺泡灌洗液，提取COPD-Exo、非COPD患者肺泡灌洗液来源的外泌体(ctrl-bronchoalveolar lavage fluid-exosome，Ctrl-Exo)，使用电镜和Western blot进行鉴定。使用茜素红染色、qRT-PCR检测COPD-Exo干预成骨细胞后成骨分化水平的改变。对GEO数据库数据集(GSE218571)中COPD-Exo的微小RNA(microRNA，miRNA)表达谱进行生物信息学分析，获得COPD-Exo中差异表达的miRNA，使用Antagomir阻碍MircoRNA功能，明确具有成骨分化调控功能的miRNA，使用Targetscan软件预测miRNA的下游靶基因并进行验证。
结果
2
COPD患者和非COPD患者的肺泡灌洗液内均可提取出外泌体。茜素红染色及PCR结果表明，COPD-Exo可以抑制人hFOB 1.19成骨细胞的成骨分化(
P
＜0.05)。生物信息学分析显示，COPD-Exo中miR-223-3p表达水平显著上调。使用Antagomir阻碍miR-223-3p可减轻COPD-Exo对于人hFOB 1.19成骨细胞的成骨分化抑制(
P
＜0.05)。Targetscan预测和Western blot结果显示miR-223-3p可能靶向抑制成骨分化相关因子FOXO3的表达(
P
＜0.05)。
结论
2
COPD-Exo可能通过miR-223-3p抑制人hFOB 1.19成骨细胞的成骨分化，该过程与miR-223-3p抑制FOXO3表达有关。
Abstract
Objective
2
To explore the role and mechanism of the exosomes derived from bronchoalveolar lavage fluid (BALF) of patients with chronic obstructive pulmonary disease (COPD-Exo) in regulating osteoblast differentiation.
Methods
2
A total of 6 COPD patients and 6 non-COPD patients admitted in our hospital in June 2023 were recruited
and their BALF samples were collected during the process. COPD-Exo and exosomes from the non-COPD patients (Ctrl-Exo) were extracted and identified by electron microscopy and Western blotting. Alizarin red staining and qRT-PCR were used to detect the differences in osteoblast differentiation after COPD-Exo and Ctrl-Exo intervention. Bioinformatics analysis was performed on the microRNA (miRNA) expression profiles of COPD-Exo and Ctrl-Exo in the GEO database (GSE218571) to obtain differentially expressed miRNAs. Antagomir was used to block mircoRNA function
and the miRNAs with osteogenic differentiation regulatory function were identified. Targetscan software was used to predict the downstream target genes of the miRNAs
and then these miRNAs were verified.
Results
2
Exo could be extracted from BALF of both COPD and non-COPD patients. Alizarin red staining and qRT-PCR results showed that COPD-Exo inhibited the osteogenic differentiation of human hFOB 1.19 osteoblasts (
P
<
0.05). Bioinformatics analysis indicated that the expression level of miR-223-3p was significantly up-regulated in COPD-Exo. Blocking miR-223-3p with Antagomir could alleviate the osteogenic differentiation of human hFOB 1.19 osteoblasts inhibited by COPD-Exo (
P
<
0.05). Targetscan prediction revealed that miR-223-3p may target and inhibit the expression of osteogenic differentiation-related factor FOXO3 (
P
<
0.05).
Conclusion
2
COPD-Exo can inhibit the osteogenic differentiation of human hFOB 1.19 osteoblasts through miR-223-3p
which may be related to the inhibition of FOXO3 expression by miR-223-3p.
关键词
Keywords
references
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