敲除Brg1基因对小鼠肺泡Ⅱ型上皮细胞数量和功能的影响及其机制研究

阮玲瑛; 薛坤娇; 应林燕; 符州; 邹文静

doi:10.16016/j.1000-5404.201908160

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敲除Brg1基因对小鼠肺泡Ⅱ型上皮细胞数量和功能的影响及其机制研究

基础医学 | 更新时间：2026-01-09

- 敲除Brg1基因对小鼠肺泡Ⅱ型上皮细胞数量和功能的影响及其机制研究
- Effect of Brahma-related gene 1 knockout on number and function of type Ⅱ alveolar epithelial cells in mice
- 第三军医大学学报 2020年42卷第7期页码：656-663
- 作者机构：
  
  1.400014 重庆，重庆医科大学附属儿童医院儿科研究所, 儿童发育疾病研究教育部重点实验室, 国家儿童健康与疾病临床医学研究中心, 儿童发育重大疾病国家国际科技合作基地
  2.儿科学重庆市重点实验室
- 作者简介：
  
  [邹文静，E-mail: lucky_121212@163.com ZOU Wenjing, E-mail: lucky_121212@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81670018)
- DOI：10.16016/j.1000-5404.201908160
  中图分类号： R322.35;R329.28;R394.3
- 收稿：2019-08-21，
  
  修回：2019-10-16，
  
  纸质出版：2020-04-15
- 稿件说明：
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阮玲瑛, 薛坤娇, 应林燕, 等. 敲除Brg1基因对小鼠肺泡Ⅱ型上皮细胞数量和功能的影响及其机制研究[J]. 第三军医大学学报, 2020,42(7):656-663.

Lingying RUAN, Kunjiao XUE, Linyan YING, et al. Effect of Brahma-related gene 1 knockout on number and function of type Ⅱ alveolar epithelial cells in mice[J]. Journal of Third Military Medical University, 2020, 42(7): 656-663.
阮玲瑛, 薛坤娇, 应林燕, 等. 敲除Brg1基因对小鼠肺泡Ⅱ型上皮细胞数量和功能的影响及其机制研究[J]. 第三军医大学学报, 2020,42(7):656-663. DOI： 10.16016/j.1000-5404.201908160.

Lingying RUAN, Kunjiao XUE, Linyan YING, et al. Effect of Brahma-related gene 1 knockout on number and function of type Ⅱ alveolar epithelial cells in mice[J]. Journal of Third Military Medical University, 2020, 42(7): 656-663. DOI： 10.16016/j.1000-5404.201908160.

摘要

目的

研究染色质重构复合物核心催化亚基(Brahma-related gene 1，Brg1)对C57BL/6小鼠Ⅱ型肺泡上皮细胞数量和功能的影响及其可能的机制。

方法

取6~8周龄雌性野生型C57BL/6小鼠(Wild type

WT)及Ⅱ型肺泡上皮细胞(alveolar epithelial cell Ⅱ

AECⅡs)上特异性敲除Brg1的C57BL/6小鼠(Brg1

fl/fl

)作为实验对象，每组选取8只小鼠，收取标本，利用免疫组化及免疫荧光分析肺组织中表面活性物质相关蛋白C(surfactant-associated protein C

SFTPC

)阳性的细胞率，流式细胞术分析小鼠肺组织中表面活性物质蛋白C前体蛋白(prosurfactant protein C

pro SP-C)阳性的细胞(即AECⅡs)数量。Western blot检测各组小鼠肺组织中pro SP-C、SFTPC、表面活性物质相关蛋白A(surfactant-associated protein A

SFTPA)、表面活性物质相关蛋白D(surfactant-associated protein D

SFTPD)的表达水平。Western blot检测细胞周期蛋白D1(CyclinD1)表达及增殖相关信号转导途径中关键蛋白EGFR、p-EGFR、PI3K、p-PI3K、AKT、p-AKT及NF-κB p65的活化水平。

结果

抗SFTPC免疫组化、免疫荧光显示其在肺泡上皮表达较WT小鼠增多(

0.05)；流式细胞术显示Brg1

fl/fl

组小鼠肺组织中pro SP-C

的细胞百分比较WT小鼠明显增高(

0.05)，Western blot检测结果显示AECⅡs的功能性蛋白pro SP-C、SFTPC、SFTPA及SFTPD在Brg1

fl/fl

小鼠中明显增多；Brg1

fl/fl

组小鼠肺组织中NF-κB p65及周期蛋白CyclinD1的表达增加，且p-EGFR/EGFR、p-PI3K/PI3K、p-AKT/AKT百分比显著提高，且差异均有统计学意义(

0.05)。

结论

特异性敲除C57BL/6小鼠Ⅱ型肺泡上皮细胞中的Brg1可促使AECⅡs数量和分泌功能的增加，其可能机制与敲除Brg1促进EGFR/PI3K/AKT/NF-κB信号通路的激活，且与增殖周期蛋白CyclinD1表达的增加有关。

Abstract

Objective

To investigate the effect of brahma-related gene 1 (Brg1) knockout on the number and function of type Ⅱ alveolar epithelial cells (AECⅡs) in C57BL/6 mice and explore the possible mechanisms.

Methods

Eight female wild-type (WT) C57BL/6 mice (6 to 8 weeks old) and 8 C57BL/6 mice with Brg1 gene knockout in type Ⅱ alveolar epithelial cells (Brg1

fl/fl

mice) were examined with immunohistochemistry and immunofluorescence assay for the number of surfactant-associated protein C (SFTPC)-positive cells in the pulmonary alveoli. The percentage of prosurfactant protein C (pro SP-C) positive cells in the lung tissues of the mice was analyzed with flow cytometry. Western blotting was used to detect the expression levels of pro SP-C

SFTPC

SFTPA and SFTPD in the lung tissues. The expression levels of cyclin D1 and the key proteins EGFR

p-EGFR

PI3K

p-PI3K

AKT

p-AKT

and NF-κB p65 in the proliferation-related signaling pathway were also detected with Western blotting.

Results

The results of immunohistochemistry and immunofluorescence assay revealed a significantly greater number of SFTPC-positive cells in Brg1

fl/fl

mice than in the WT mice (

0.05). Flow cytometry showed that the percentage of pro SP-C positive cells increased significantly in the lung tissues of Brg1

fl/fl

mice (

0.05). The expression levels of the functional proteins in AECⅡs

including proSP-C

SFTPC

SFTPA and SFTPD

were all significantly increased in Brg1

fl/fl

mice (

0.05); Brg1 knockout significantly promoted the expression of NF-κB p65 and CyclinD1 (

0.05) and increased the ratios of p-EGFR/EGFR

p-PI3K/PI3K and p-AKT/AKT in the lung tissue of the mice (

0.05).

Conclusion

Specific knockout of Brg1 in AECⅡs of C57BL/6 mice increases the number and enhances the functionality of AECⅡs possibly due to the activation of EGFR/PI3K/AKT/NF-κB signaling pathways and an increased protein expression of CyclinD1.

关键词

Keywords

references

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