Role of methyl-CpG-binding domain protein 2 in regulating aquaporin-5 expression in human submandibular gland cells <italic style="font-style: italic">in vitro</italic>

Zhonglin CHEN; Minhai NIE; Ying JIA; Yanyan YE; Shuting DENG; Wei HUANG; Xuqian LIU

doi:10.16016/j.1000-5404.201811163

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Role of methyl-CpG-binding domain protein 2 in regulating aquaporin-5 expression in human submandibular gland cells in vitro

Basic Medicine | 更新时间：2026-01-09

- Role of methyl-CpG-binding domain protein 2 in regulating aquaporin-5 expression in human submandibular gland cells in vitro
- Journal of Third Military Medical University Vol. 41, Issue 10, Pages: 955-960(2019)
- 作者机构：
  
  1.646000 四川泸州；西南医科大学附属口腔医院牙周黏膜科
  2.400038 重庆，陆军军医大学(第三军医大学)第一附属医院口腔科
- 作者简介：
- 基金信息：
  
  Science Research Project of Southwest Medical University(2017-ZRQN-083);Scientific Research Project of Sichuan Medical Association(S17073);Doctoral Research Initiation Fund for City-School-Stomatological Hospital Joint(西南医大科[2018] 6号-2)
- DOI：10.16016/j.1000-5404.201811163
  CLC： R329.2;R394.2;R781.7
- Received：12 November 2018，
  
  Revised：2019-01-02，
  
  Published：30 May 2019
- 稿件说明：
移动端阅览
Zhonglin CHEN, Minhai NIE, Ying JIA, et al. Role of methyl-CpG-binding domain protein 2 in regulating aquaporin-5 expression in human submandibular gland cells in vitro[J]. Journal of Third Military Medical University, 2019, 41(10): 955-960.
DOI：

Zhonglin CHEN, Minhai NIE, Ying JIA, et al. Role of methyl-CpG-binding domain protein 2 in regulating aquaporin-5 expression in human submandibular gland cells in vitro[J]. Journal of Third Military Medical University, 2019, 41(10): 955-960. DOI： 10.16016/j.1000-5404.201811163.

摘要

目的

探讨人下颌下腺细胞(human submandibular gland，HSG)中，甲基化CpG结合域蛋白2(menthyl-CpG-binding domain protein 2，MBD2)是否影响水通道蛋白调控-5(aquaporin 5，AQP5)的表达，为干燥综合征(Sjögren’s syndrome，SS)的治疗提供新的思路。

方法

体外培养HSG细胞，通过AQP5的启动子质粒与MBD2-siRNA或MBD2过表达质粒共转染HSG细胞，利用双荧光素酶报告基因系统检测AQP5启动子的活性；再用MBD2-siRNA或MBD2过表达质粒转染HSG细胞，利用RT-PCR及Western blot技术检测MBD2和AQP5在mRNA水平及蛋白水平上表达的改变。

结果

① MBD2-siRNA转染HSG细胞后，AQP5 pro活性分别上调49.30%和58.35%(

0.05)

AQP5在mRNA水平上表达上调35.40%和471.19%(

0.05)，蛋白水平上调56.16%和82.78% (

0.05)。结果表明：MBD2-siRNA转染HSG细胞后，AQP5启动子活性增加，AQP5表达上调。②MBD2过表达质粒转染HSG细胞后，AQP5 pro活性分别下调25.36%、30.52%及34.78%(

0.05)

AQP5在mRNA水平上表达下调8.06%和87.60%(过表达质粒1 μg组与对照组相比无统计学意义，

0.05；过表达质粒2 μg组与对照组相比差异有统计学意义

0.05)，在蛋白水平上表达下调44.50%和53.18%(

0.05)。实验结果表明：MBD2转染ASG细胞后，AQP5启动子活性降低，AQP5的表达下降(

0.05)。

结论

MBD2可以下调AQP5的表达，这与SS患者的AQP5表达趋势一致，因此MBD2可能成为干燥综合征基因治疗的新靶点。

Abstract

Objective

To study whether methyl-CpG-binding domain protein 2 (MBD2) affects the expression of aquaporin-5 (AQP5) in human submandibular gland (HSG) cells

thereby exploring a new approach to treatment of Sjögren's syndrome (SS).

Methods

Cultured HSG cells were co-transfected with the promoter plasmid of AQP5 and the plasmids for MBD2 silencing (MBD2-siRNA) or overexpression. Luciferase report gene assay was used to detect the transcription level of AQP5 in the transfected cells. In HSG cells transfected with the MBD2-siRNA or the overexpression plasmids

RT-qPCR and Western blotting were performed to analyze the changes in MBD2 and AQP5 expression at both the mRNA and protein levels.

Results

Transfection of the cells with the 2 MBD2-siRNAs respectively increased AQP5 promoter activity by 49.30% and 58.35% (

0.05) and obviously upregulated the expression of AQP5 by 35.40% and 471.19% at the mRNA level (

0.05) and by 56.16% and 82.78% at the protein level (

0.05). Transfection of the cells with the three MBD2 overexpression plasmids resulted in significantly lowered AQP5 promoter activity by 25.36%

30.52% and 34.78% (

0.05); while 1 plasmid produced no significant changes in AQP5 mRNA level

the other 2 plasmids significantly down-regulated AQP5 expression by 8.06% and 87.60% at the mRNA level (

0.05) and by 44.50% and 53.18% at the protein level (

0.05).

Conclusion

MBD2 could down-regulats the expression of AQP5 in HSG cells

in vitro

which consistent with the AQP5 expression about patients with SS

and this finding indentifies MBD2 as a potential target in gene therapy for the syndrome.

关键词

Keywords

references

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Role of methyl-CpG-binding domain protein 2 in regulating aquaporin-5 expression in human submandibular gland cells in vitro

DOI：10.16016/j.1000-5404.201811163

摘要

Abstract

关键词

Keywords

references